METHOD FOR DIAGNOSING HEMATOLOGICAL DISORDERS

Abstract

Disclosed is a method for the diagnosis, and/or the classification, of a hematological disorder, including the steps of: a). measuring, the expression level of at least the genes of a sub-group of 6 genes, b). comparing the expression level of each genes measured in step a)., with the expression level of the same genes in healthy control sample, and c). determining the status of the biological sample.

Description

[0001] The present invention relates to a method for diagnosing hematological disorders in a patient.

[0002] The great quantity of hematopoietic cells and the many stages of differentiation through which they pass further complicate the classification of the neoplasis originating from this type of cells. Despite the efforts to establish a classification based on "real" entities, some of the categories are ambiguous and in many cases contain very heterogeneous groups as regards a response to therapy of clinical course. This heterogeneity is that responsible for, on the one hand, the incessant search for markers capable of differentiating some behaviours from others and, on the other hand, that the disputed classification of this type of neoplasia is subjected to continuous revisions.

[0003] An ideal classification system should be precise, reproducible, easy to use and should especially have biological and clinical significance (Chan W C et al., Croat Med J. 2005; 46:349-59). The current diagnosis systems and the classification of the hematological neoplasias are based on the recognition of histological and morphological, immunophenotypical and cytogenetic characteristics and study of a molecular marker with prognostic value. However, in some of the diagnostic categories defined in this way, the following is observed:

[0004] A marked heterogeneous therapy response: within the same disease, patients either reach full remission, or partial remission, or do not respond, or relapse after a certain therapy. The capacity to predict a response is especially important in this type of neoplasias since the transplant of stem cells is an effective but toxic alternative response. The capacity to determine which patients would respond to a conventional therapy before giving it may be beneficial to be able to apply the most effective treatment to each patient.

[0005] A variable clinical behaviour: within this category for some patients the disease is going to remain stable for long periods of time and that are not going to need therapy and whereas for others the disease is going to progress rapidly requiring aggressive therapy.

[0006] These variations point to the existence of molecular heterogeneity within the diagnostic categories, differences which the conventional methods of diagnosis are not capable of determining and hence, the search for new forms of analysis which provide a greater resolution in the characterization of this type of neoplasias.

[0007] In this line, the use of expression arrays have demonstrated being effective not only in deciphering the biological and clinical diversity which is found in many tumours, but in understanding the biological and pathological processes which affect many symptoms and, in particular, the hematopoietic system. The expression arrays are ordered arrays of sequences associated to a solid support, complementary to mRNA or to its corresponding cDNA or cRNA, which allow the analysis of the differential expression of hundreds or thousands of genes simultaneously. One of the supports to which they are frequently bound is to rectangular fragments of glass similar to slides, a format which is frequently alluded to by the terms microarray, biochip or, simply, chip. Their use is becoming increasingly frequent for the diagnosis of various diseases or for the evolution of the evaluation of the susceptibility of suffering from them.

[0008] In 1999, the Golub group published one of the first articles referring to the role of arrays in the classification of hematological neoplasias (Golub T R et al, Science. 1999; 286:531-7). An array with 6817 genes represented was used for the study of expression profiles in acute myeloid leukemia (AML) and acute lymphoid leukemia (ALL). A group of 50 genes was selected with the capacity of predicting the type of leukemia (class predictor) and they were used to classify a group of unknown samples in the correct categories. The study of the expression of these 50 genes is sufficient for the classification of a sample of AML or ALL. Despite the fact that the distinction between AML and ALL is well established with the current diagnostic methods, the study revealed the existence of specific expression patterns associated with each type of acute leukemia and proved the use.

[0009] The European patent application EP1947194 has proposed the use of specific oligonucleotides for diagnosing chronic lymphoid leukemia (CLL) diseases.

[0010] The international application WO 2005/080601 discloses methods of genetic analysis for the classification, diagnosis and prognosis of acute myeloid leukemia (AML). This application provides a method for differentiating AML subtypes, but never provides methods for the discrimination between preleukemic and leukemic states.

[0011] The international application WO 2006/125195 discloses a group of 24 genes whose expression allows to classify a sample as myelodysplastic syndrome (MDS), AML, or not diseased. However, this document does not provide a method for classifying different grade of MDS.

[0012] So there is a need to provide a new method for diagnosing, classifying the malignant and premalignant states of cancer.

[0013] Oxidative stress is generally defined as an imbalance between the generation of reactive oxygen species (ROS) and impaired antioxidant defense systems. It has long been known to be involved in the pathophysiology of cancer.

[0014] High level of ROS produced either endogenously or exogenously can attack lipids, proteins, and nucleic acids simultaneously in living cells. This has led to cells developing various antioxidant defense mechanisms to both prevent the excessive formation of ROS and limit their harmful effects. The appropriate redox balance is maintained via the combined action of antioxidant enzymes.

[0015] Myelodysplastic syndroma and acute leukemia are characterized by a pathological hematopoieisis. ROS certainly plays an important role in human hematopoiesis. For example, it is well established in murine models that ROS-induced p38 MAPK activation is crucial in hematopoiesis and increased ROS levels are required to trigger hematopoietic stem cells (HSC) exit from quiescence and to drive maturation and differentiation. Moreover, a high ROS level induces a perturbation in the self-renewal activity of HSC. It is now well established that the progression of normal cells to neoplastic transformation results from the accumulation of mutations in genes that control cellular proliferation, survival, and differentiation. Approximately, 30% myelodysplastic syndroma cases progress to acute leukemia. Indirect evidences suggest a role for oxidant DNA damage in the pathogenesis of myelodysplasia. Moreover, the flow cytometric quantification of ROS in bone marrow cells from myelodysplastic and leukemic patients reveals an increase in ROS level in all cases.

[0016] So, studying the antioxidant response in premalignant and malignant state seems to be a good start for providing a new useful method.

[0017] The cancer stem cell (CSC) hypothesis suggests that a subset of cells within a tumor has the ability to recapitulate the generation of a continuously growing tumor (Clarke M F et al, Cancer Research. 2006; 66:9339-44). CSCs are best described in human in which the rare so-called leukemia stem cells (L-HSCs) can be prospectively isolated and shown to transmit the disease when introduced into immuno-compromised mice (Lap/dot T et al, Nature. 1994; 645-8). Cells which do not share this phenotype often represent the bulk of the leukemic clone, but fail to transmit the disease upon transplantation. The early work on L-HSC has now been extended to a list of tumors which is rapidly expanding (Bomken S, Br. J. Cancer. 2010; 103.439-45). Because they appear to be resistant to drugs that are commonly used to treat leukemia in humans, L-HSCs may be responsible for relapse in some patients (Ishikawa F, Nat. Biotechnol 2007; 25:1315-21). Genes which are functionally significant for L-HSC expansion may therefore represent the ultimate therapeutic targets. This raises the possibility that other ROS scavenging systems are of regulatory importance in other cancer stem cells. Indeed a low ROS level in breast CSCs has recently been reported, where it was associated with increased expression of the glutathione biosynthesis genes (Diehn M, Nature. 2009; 458:780-3).

[0018] Therefore, one aim of the invention is to provide a new method for diagnosing cancer.

[0019] Another aim of the invention is to provide a rapid efficient method for classifying pathologic sample, which cannot be classified by other method.

[0020] Another aim of the invention is to provide a kit for the implementation of the above methods.

[0021] Another aim of the invention is to provide composition allowing the implementation of the above method.

[0022] The invention relates to a method for the diagnosis, and/or the classification, preferably in vitro, of an hematological disorder, in particular myeloid and/or lymphoid hematological disorder, preferably myeloid hematological disorder,

[0023] said method comprising the steps of:

a). measuring, from cells contained in a biological sample of a subject, preferably from blood cells or bone marrow cells containing sample, the expression level of at least the genes of a sub-group of 6 genes belonging to a set of genes chosen among a group of 24 genes, said group of 24 genes comprising or being constituted by the nucleic acid sequences SEQ ID NO:1 to 24, wherein said subject is suspected to be afflicted by an hematological disorder, in particular myeloid and/or lymphoid hematological disorder, preferably myeloid hematological disorder, said 6 genes belonging to said sub-group comprising or being constituted by the nucleic acid sequences SEQ ID NO: 1 to 6, b). comparing the expression level of each genes measured in step a)., with the expression level of the same respective genes from cells contained in a control sample preferably from blood cells or bone marrow cells containing sample, said control sample being of the same nature than said biological sample, to establish a gene expression level ratio for each genes of said sub-group, and c). determining the status of said biological sample such that if the ratio established in step b). for each genes of any combination of at least 3 genes from said sub-group is either .gtoreq.2 or .ltoreq.0.5, said biological sample is representative of an hematological disorder cells.

[0024] In other words, the step b). according to the invention consists of:

comparing the expression level of each genes measured in step a)., with the expression level of the same respective genes from cells contained in a control sample, preferably from a control sample containing blood cells or bone marrow cells, said control sample being of the same nature than said biological sample, to establish a gene expression level ratio R.sub.i between the expression level of each genes i measured in step a) and the expression level of the same respective genes i from cells contained in a control sample, for each genes of said sub-group.

[0025] The step c). according to the invention consists of determining the status of said biological sample such that if the ratio R.sub.i for each genes of any combination of 3 genes from said sub-group is

[0026] either .gtoreq.2,

[0027] or .ltoreq.0.5,

[0028] said biological sample is representative of an haematological disorder cells.

[0029] The invention is based on the unexpected observation made by the Inventors that at least 6 specific genes, i.e. the genes comprising or being constituted by the nucleic acid sequences SEQ ID NO 1-6, belonging to a group of 24 specific genes comprising or being constituted by the nucleic acid sequences SEQ ID NO 1-24 are sufficient to determine the status a of an hematological disorder.

[0030] The invention is preferably carried out with sample from patients who have not been previously treated for an hematological disorder.

[0031] These genes are specifically genes coding for enzymes involved in the detoxification of cells, in which ROS accumulate.

[0032] The natural process involved in the elimination of ROS is represented in FIG. 1.

[0033] According to the invention the group (C) of 24 genes comprises a set (B) of genes, said set comprising the subgroup (A) of 6 specific genes as defined above. The imbrications of the group/set/subgroup according to the invention are represented in FIG. 2.

[0034] The method according to the invention is thus carried out as follows:

[0035] from a sample of a patient, the nucleic acid molecules contained in said sample are extracted, preferably the RNA molecules, according to extraction methods known in the art,

[0036] the amount of specific nucleic acid molecules, corresponding to the nucleic acid molecules comprising or being constituted by at least SEQ ID NO: 1-6, is quantified, by well known techniques as illustrated hereafter,

[0037] the amount quantified in the above step is compared with the amount of the same nucleic acid molecules contained in a control sample, and a ratio is established.

[0038] Accordingly, the control sample, which is used as reference, is a sample of an healthy individual, said healthy sample being of the same nature than the sample of the patient. Advantageously, the control sample corresponds to a pool of numerous samples of different healthy individuals, i.e. the control sample represents the mean of numerous healthy individual.

[0039] As mentioned above, the sample of the patient and the control sample are of the same origin. This means that if the sample of the patient is originated from blood of the patient, the control sample is originated from blood of one or many healthy individuals. Blood, in the invention, means total blood, plasma, serum, peripheral blood mononuclear cells (PBMC) . . . .

[0040] In the same way, if the sample of the patient is originated from bone marrow, the control sample is originated from bone marrow of one or many healthy individuals.

[0041] In the invention, the biological of the patient, from whom the biological sample is used, is suspected to be afflicted by an hematological disorder, in particular myeloid and/or lymphoid hematological disorder, preferably myeloid hematological disorder.

[0042] This means that the pathological status of the patient is:

[0043] either undetermined (the pathologist does not know if said patient is afflicted by an hematological disorder),

[0044] or determined (the pathologist knows that the patient is afflicted by an hematological disorder).

[0045] This also means that the pathological status of the patient is:

[0046] first determined (the pathologist identify if the patient is afflicted by an hematological disorder, or not),

[0047] and if the patient is afflicted by an hematological disorder, said hematological disorder is classified.

[0048] If the pathological status is undetermined, then the pathologist measure the expression levels of the genes consisting of SEQ ID NO: 1-6 in a biological sample from said patient and compare said expression levels with the expression levels of the same genes (i.e. genes SEQ ID NO: 1-6) in a control sample of the same nature, as defined above, (for instance a pool of cells from healthy donors that is used as control sample or reference), in order to establish the ratios Ri, and to determine if the patient is afflicted by hematological disorder or not.

[0049] If the pathological status is determined (for instance by cytological studies), the pathologist measures the expression levels of the genes consisting of SEQ ID NO: 1-6 in a biological sample from said patient and compares said expression levels with the expression levels of the same genes (i.e. genes SEQ ID NO: 1-6) in a control sample of the same nature, as defined above, (for instance a pool of cells from healthy donors that is used as control sample or reference), in order to establish the ratios Ri, and can classify the hematological disorder according to the method of the invention.

[0050] The method according to the invention provides an easy to use, rapid and efficient process to evaluate the status of a sample identified as, or suspected to be, a sample corresponding to an hematological disorder, in particular a myeloid disorder.

[0051] If the ratio between the expression level of at least 3 genes, of the above 6 genes belonging to the above defined subgroup, of a patient sample and the expression level of the same at least 3 genes of a control sample is either .gtoreq.2 or .ltoreq.0.5, then the sample of the patient would be considered as presenting the features of a sample corresponding to an hematological disorder.

[0052] Above and hereafter, the ratio is defined as follows:

Ri=[Amount(expression level) of a gene i of SEQ ID NO:i in the patient sample]/[Amount(expression level) of a gene i of SEQ ID NO:i in the control sample]

[0053] i varying from 1 to 24.

[0054] Therefore R.sub.3 represents the ratio as defined above, relative to the gene SEQ ID NO: 3, and thus R.sub.i represents the ratio as defined above, relative to the gene SEQ ID NO: i, i varying from 1 to 24.

[0055] In the invention, "the ratio R.sub.i of each gene of any combination of at least 3 genes of the 6 genes of said subgroup is .gtoreq.2 or .ltoreq.0.5" means that the ratio R.sub.i of each gene of any combination of 3, or 4 or 5 or 6 genes is .gtoreq.2 or .ltoreq.0.5.

[0056] All the 20 combinations of 3 genes chosen among the 6 genes are listed hereafter:

SEQ ID NO: 1+SEQ ID NO: 2+SEQ ID NO: 3,

SEQ ID NO: 1+SEQ ID NO: 2+SEQ ID NO: 4,

SEQ ID NO: 1+SEQ ID NO: 2+SEQ ID NO: 5,

SEQ ID NO: 1+SEQ ID NO: 2+SEQ ID NO: 6,

SEQ ID NO: 1+SEQ ID NO: 3+SEQ ID NO: 4,

SEQ ID NO: 1+SEQ ID NO: 3+SEQ ID NO: 5,

SEQ ID NO: 1+SEQ ID NO: 3+SEQ ID NO: 6,

SEQ ID NO: 1+SEQ ID NO: 4+SEQ ID NO: 5,

SEQ ID NO: 1+SEQ ID NO: 4+SEQ ID NO: 6,

SEQ ID NO: 1+SEQ ID NO: 5+SEQ ID NO: 6,

SEQ ID NO: 2+SEQ ID NO: 3+SEQ ID NO: 4,

SEQ ID NO: 2+SEQ ID NO: 3+SEQ ID NO: 5,

SEQ ID NO: 2+SEQ ID NO: 3+SEQ ID NO: 6,

SEQ ID NO: 2+SEQ ID NO: 4+SEQ ID NO: 5,

SEQ ID NO: 2+SEQ ID NO: 4+SEQ ID NO: 5,

SEQ ID NO: 2+SEQ ID NO: 5+SEQ ID NO: 6,

SEQ ID NO: 3+SEQ ID NO: 4+SEQ ID NO: 5,

SEQ ID NO: 3+SEQ ID NO: 4+SEQ ID NO: 6,

SEQ ID NO: 3+SEQ ID NO: 5+SEQ ID NO: 6 and

SEQ ID NO: 4+SEQ ID NO: 5+SEQ ID NO: 6.

[0057] All the 15 combinations of 4 genes among the 6 genes are the following ones:

SEQ ID NO: 1+SEQ ID NO: 2+SEQ ID NO: 3+SEQ ID NO: 4,

SEQ ID NO: 1+SEQ ID NO: 2+SEQ ID NO: 3+SEQ ID NO: 5,

SEQ ID NO: 1+SEQ ID NO: 2+SEQ ID NO: 3+SEQ ID NO: 6,

SEQ ID NO: 1+SEQ ID NO: 3+SEQ ID NO: 4+SEQ ID NO: 5,

SEQ ID NO: 1+SEQ ID NO: 3+SEQ ID NO: 4+SEQ ID NO: 6,

SEQ ID NO: 1+SEQ ID NO: 2+SEQ ID NO: 4+SEQ ID NO: 5,

SEQ ID NO: 1+SEQ ID NO: 2+SEQ ID NO: 4+SEQ ID NO: 6,

SEQ ID NO: 1+SEQ ID NO: 2+SEQ ID NO: 5+SEQ ID NO: 6,

SEQ ID NO: 1+SEQ ID NO: 2+SEQ ID NO: 3+SEQ ID NO: 5,

SEQ ID NO: 1+SEQ ID NO: 2+SEQ ID NO: 3+SEQ ID NO: 6,

SEQ ID NO: 2+SEQ ID NO: 3+SEQ ID NO: 4+SEQ ID NO: 5,

SEQ ID NO: 2+SEQ ID NO: 3+SEQ ID NO: 4+SEQ ID NO: 6,

SEQ ID NO: 2+SEQ ID NO: 3+SEQ ID NO: 5+SEQ ID NO: 6,

SEQ ID NO: 2+SEQ ID NO: 4+SEQ ID NO: 5+SEQ ID NO: 6 and

SEQ ID NO: 3+SEQ ID NO: 4+SEQ ID NO: 5+SEQ ID NO: 6.

[0058] All the 6 combinations of 5 genes among 6 genes are the following ones:

SEQ ID NO: 1+SEQ ID NO: 2+SEQ ID NO: 3+SEQ ID NO: 4+SEQ ID NO: 5,

SEQ ID NO: 1+SEQ ID NO: 2+SEQ ID NO: 3+SEQ ID NO: 4+SEQ ID NO: 6,

SEQ ID NO: 1+SEQ ID NO: 2+SEQ ID NO: 3+SEQ ID NO: 5+SEQ ID NO: 6,

SEQ ID NO: 1+SEQ ID NO: 2+SEQ ID NO: 4+SEQ ID NO: 5+SEQ ID NO: 6,

SEQ ID NO: 1+SEQ ID NO: 3+SEQ ID NO: 4+SEQ ID NO: 5+SEQ ID NO: 6 and

SEQ ID NO: 2+SEQ ID NO: 3+SEQ ID NO: 4+SEQ ID NO: 5+SEQ ID NO: 6.

[0059] Finally, the combination of the six genes is SEQ ID NO: 1+SEQ ID NO: 2+SEQ ID NO: 3+SEQ ID NO: 4+SEQ ID NO: 5+SEQ ID NO: 6.

[0060] According to the invention, a sample of a patient wherein the ratio R.sub.i of each gene belonging to any combination of least 3 genes of the subgroup of 6 genes is .gtoreq.2 or .ltoreq.0.5 will be considered as a sample corresponding to an hematological disorder.

[0061] For instance, if the combination SEQ ID NO: 1+SEQ ID NO: 2+SEQ ID NO: 3 is studied, if the respective ratios R.sub.1, R.sub.2 and R.sub.3 as described above are as follows:

R1.gtoreq.2 and R2.gtoreq.2 and R3.gtoreq.2, or

R1.ltoreq.0.5 and R2.gtoreq.2 and R3.gtoreq.2, or

R1.gtoreq.2 and R2.ltoreq.0.5 and R3.gtoreq.2, or

R1.gtoreq.2 and R2.gtoreq.2 and R3.ltoreq.0.5, or

R1.ltoreq.0.5 and R2.ltoreq.0.5 and R3.gtoreq.2, or

R1.ltoreq.0.5 and R2.gtoreq.2 and R3.ltoreq.0.5, or

R1.gtoreq.2 and R2.ltoreq.0.5 and R3.ltoreq.0.5, or

R1.ltoreq.0.5 and R2.ltoreq.0.5 and R3.ltoreq.0.5,

[0062] then the sample of the patient in which the ratios are calculated will be considered as a sample corresponding to an hematological disorder.

[0063] The above example applies mutatis mutandis to the combinations of at least 3 genes mentioned above.

[0064] Consequently, if the combination SEQ ID NO: 1+SEQ ID NO: 2+SEQ ID NO: 3+SEQ ID NO: 4 is studied, 4 combinations of 3 genes exist:

SEQ ID NO: 1+SEQ ID NO: 2+SEQ ID NO: 3,

SEQ ID NO: 1+SEQ ID NO: 2+SEQ ID NO:4,

SEQ ID NO: 1+SEQ ID NO: 3+SEQ ID NO: 4, and

SEQ ID NO: 2+SEQ ID NO: 3+SEQ ID NO: 4.

[0065] Therefore, if the respective ratios R.sub.1, R.sub.2, R.sub.3 and R.sub.4 are as follows:

[0066] for combination 1:

R1.gtoreq.2 and R2.gtoreq.2 and R3.gtoreq.2, or

R1.ltoreq.0.5 and R2.gtoreq.2 and R3.gtoreq.2, or

R1.gtoreq.2 and R2.ltoreq.0.5 and R3.gtoreq.2, or

R1.gtoreq.2 and R2.gtoreq.2 and R3.ltoreq.0.5, or

R1.ltoreq.0.5 and R2.ltoreq.0.5 and R3.gtoreq.2, or

R1.ltoreq.0.5 and R2.gtoreq.2 and R3.ltoreq.0.5, or

R1.gtoreq.2 and R2.ltoreq.0.5 and R3.ltoreq.0.5, or

R1.ltoreq.0.5 and R2.ltoreq.0.5 and R3.ltoreq.0.5,

[0066]

[0067] for combination 2

R1.gtoreq.2 and R3.gtoreq.2 and R4.gtoreq.2, or

R1.ltoreq.0.5 and R3.gtoreq.2 and R4.gtoreq.2, or

R1.gtoreq.2 and R3.ltoreq.0.5 and R4.gtoreq.2, or

R1.gtoreq.2 and R3.gtoreq.2 and R4.ltoreq.0.5, or

R1.ltoreq.0.5 and R3.ltoreq.0.5 and R4.gtoreq.2, or

R1.ltoreq.0.5 and R3.gtoreq.2 and R4.ltoreq.0.5, or

R1.gtoreq.2 and R3.ltoreq.0.5 and R4.ltoreq.0.5, or

R1.ltoreq.0.5 and R3.ltoreq.0.5 and R4.ltoreq.0.5,

[0067]

[0068] for combination 3,

R1.gtoreq.2 and R2.gtoreq.2 and R4.gtoreq.2, or

R1.ltoreq.0.5 and R2.gtoreq.2 and R4.gtoreq.2, or

R1.gtoreq.2 and R2.ltoreq.0.5 and R4.gtoreq.2, or

R1.gtoreq.2 and R2.gtoreq.2 and R4.ltoreq.0.5, or

R1.ltoreq.0.5 and R2.ltoreq.0.5 and R4.gtoreq.2, or

R1.ltoreq.0.5 and R2.gtoreq.2 and R4.ltoreq.0.5, or

R1.gtoreq.2 and R2.ltoreq.0.5 and R4.ltoreq.0.5, or

R1.ltoreq.0.5 and R2.ltoreq.0.5 and R4.ltoreq.0.5,

[0068]

[0069] for combination 4

R2.gtoreq.2 and R3.gtoreq.2 and R4.gtoreq.2, or

R2.ltoreq.0.5 and R3.gtoreq.2 and R4.gtoreq.2, or

R2.gtoreq.2 and R3.ltoreq.0.5 and R4.gtoreq.2, or

R2.gtoreq.2 and R3.gtoreq.2 and R4.ltoreq.0.5, or

R2.ltoreq.0.5 and R3.ltoreq.0.5 and R4.gtoreq.2, or

R2.ltoreq.0.5 and R3.gtoreq.2 and R4.ltoreq.0.5, or

R2.gtoreq.2 and R3.ltoreq.0.5 and R4.ltoreq.0.5, or

R2.ltoreq.0.5 and R3.ltoreq.0.5 and R4.ltoreq.0.5,

[0070] then the sample of the patient in which the ratios are calculated will be considered as a sample corresponding to an hematological disorder.

[0071] In the invention, the genes for which the expression level is measured are represented by the RNA molecules obtained by the transcription of said genes. The transcription process is well known in the art.

[0072] Therefore, the invention relates to a process as defined above, in which the expression level of the above genes is measured by determining the amount of RNA molecules that are the products of the transcription of said genes, i.e. which are the products of the expression of said genes.

[0073] Some genes in the invention are able to express many variants, i.e. many RNA molecules that differ in their sequences. Theses variants generally differ in there sequence after alternative splicing, said alternative splicing having as consequence to add, to delete and/or to modify one or more parts of the nucleic acid sequence contained in the gene in the resulting RNA molecule. The skilled person knows the mechanisms of alternative splicing.

[0074] Therefore some genes according to the invention can express more than one RNA molecule, and provide variants.

[0075] The PRDX gene is able to express 3 different variants: the first variant comprising or consisting of the nucleic acid sequence SEQ ID NO: 5, a second variant comprising or consisting of the nucleic acid sequence SEQ ID NO: 73 and a third variant comprising or consisting of the nucleic acid sequence SEQ ID NO: 74.

[0076] The SOD2 gene is able to express 3 different variants: the first variant comprising or consisting of the nucleic acid sequence SEQ ID NO: 7, a second variant comprising or consisting of the nucleic acid sequence SEQ ID NO: 75 and a third variant comprising or consisting of the nucleic acid sequence SEQ ID NO: 76.

[0077] The GSR gene is able to express 4 different variants: the first variant comprising or consisting of the nucleic acid sequence SEQ ID NO: 8, a second variant comprising or consisting of the nucleic acid sequence SEQ ID NO: 77, a third variant comprising or consisting of the nucleic acid sequence SEQ ID NO: 78 and a fourth variant comprising or consisting of the nucleic acid sequence SEQ ID NO: 79.

[0078] The GLRX gene is able to express 2 different variants: the first variant comprising or consisting of the nucleic acid sequence SEQ ID NO: 9 and a second variant comprising or consisting of the nucleic acid sequence SEQ ID NO: 80.

[0079] The PDRX5 gene is able to express 3 different variants: the first variant comprising or consisting of the nucleic acid sequence SEQ ID NO: 11, a second variant comprising or consisting of the nucleic acid sequence SEQ ID NO: 81 and a third variant comprising or consisting of the nucleic acid sequence SEQ ID NO: 82.

[0080] The GPX4 gene is able to express 3 different variants: the first variant comprising or consisting of the nucleic acid sequence SEQ ID NO: 14, and a second variant comprising or consisting of the nucleic acid sequence SEQ ID NO: 83.

[0081] The PDRX5 gene is able to express 3 different variants: the first variant comprising or consisting of the nucleic acid sequence SEQ ID NO: 16, a second variant comprising or consisting of the nucleic acid sequence SEQ ID NO: 84 and a third variant comprising or consisting of the nucleic acid sequence SEQ ID NO: 85.

[0082] Thus, according to the invention, the measure of the expression level of the gene comprising or being constituted by SEQ ID NO: 5, can be evaluated by the measure of the expression level of the gene comprising or being constituted by SEQ ID NO: 73 or SEQ ID NO: 74.

[0083] In the same manner, the measure of the expression level of the gene comprising or being constituted by SEQ ID NO: 7, can be evaluated by the measure of the expression level of the gene comprising or being constituted by SEQ ID NO: 75 or SEQ ID NO: 76.

[0084] Moreover, the measure of the expression level of the gene comprising or being constituted by SEQ ID NO: 8, can be evaluated by the measure of the expression level of the gene comprising or being constituted by SEQ ID NO: 77, SEQ ID NO: 78 or SEQ ID NO: 79.

[0085] Moreover, the measure of the expression level of the gene comprising or being constituted by SEQ ID NO: 9, can be evaluated by the measure of the expression level of the gene comprising or being constituted by SEQ ID NO: 80.

[0086] Moreover, the measure of the expression level of the gene comprising or being constituted by SEQ ID NO: 11, can be evaluated by the measure of the expression level of the gene comprising or being constituted by SEQ ID NO: 81 or SEQ ID NO: 82.

[0087] Moreover, the measure of the expression level of the gene comprising or being constituted by SEQ ID NO: 14, can be evaluated by the measure of the expression level of the gene comprising or being constituted by SEQ ID NO: 83.

[0088] Moreover, the measure of the expression level of the gene comprising or being constituted by SEQ ID NO: 16, can be evaluated by the measure of the expression level of the gene comprising or being constituted by SEQ ID NO: 84 or SEQ ID NO: 85.

[0089] Therefore, as disclosed before and hereafter in the invention, the genes comprising or being constituted by the following sequences: SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 14 and SEQ ID NO: 16 can be replaced by their respective variants, as defined above.

[0090] The genes used according to the invention are represented in the following table 1:

TABLE-US-00001 TABLE 1 Gene name SEQ ID GPX3 SEQ ID NO: 1 GPX1(1) SEQ ID NO: 2 GLRX2(2) SEQ ID NO: 3 CAT SEQ ID NO: 4 PRDX(1-2-3) SEQ ID NO: 5 SEQ ID NO: 73 SEQ ID NO: 74 PRDX5(2) SEQ ID NO: 6 SOD2(1-2-3) SEQ ID NO: 7 SEQ ID NO: 75 SEQ ID NO: 76 GSR (1-2-3-4) SEQ ID NO: 8 SEQ ID NO: 77 SEQ ID NO: 78 SEQ ID NO: 79 GLRX(1-2) SEQ ID NO: 9 SEQ ID NO: 80 PRDX2(1) SEQ ID NO: 10 PRDX5(1-3) SEQ ID NO: 11 SEQ ID NO: 81 SEQ ID NO: 82 SOD1 SEQ ID NO: 12 TXN SEQ ID NO: 13 PRDX3(1-2) SEQ ID NO: 14 SEQ ID NO: 83 GPX7 SEQ ID NO: 15 GPX4(1-2-3) SEQ ID NO: 16 SEQ ID NO: 84 SEQ ID NO: 85 TXN2 SEQ ID NO: 17 PRDX4 SEQ ID NO: 18 GPX1(2) SEQ ID NO: 19 GLRX3 SEQ ID NO: 20 PRDX2(3) SEQ ID NO: 21 PRDX6 SEQ ID NO: 22 GLRX5 SEQ ID NO: 23 GLRX2(1) SEQ ID NO: 24

[0091] Table 1 represents SEQ ID of the genes, and the variant when they exist, used in the invention.

[0092] According to the invention, hematological disorders correspond to disorders which primarily affect the blood. In particular, hematological disorders according to the invention encompass all cytopenias (anemia: decrease in red blood cell count or hemoglobin, thrombopenias: decrease in blood platelet count, and leukopenias: decrease in leukocyte count) whatever the mechanism such as hemoglobinopathies and myelodysplastic syndrome, myeloproliferative disorders (increased numbers of myeloid cells or myelofibrosis, including chronic myeloid leukemia, polycythemia vera, essential thrombocythemia, idiopathic myelofibrosis), lymphoproliferative disorders (increased numbers of lymphoid cells, including chronic lymphocytic leukemia, lymphomas, myeloma, plasmacytoma), acute leukemias and coagulopathies (disorders of bleeding and coagulation).

[0093] In one advantageous embodiment, the invention relates to a method as defined above, wherein if the ratio established in step b). is

[0094] .ltoreq.0.3, for the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 1, and

[0095] .gtoreq.3.0, for the genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 2 and 3, then said biological sample is representative of an acute myeloid leukemia.

[0096] In other words, an embodiment of the invention relates to a method as defined above, wherein if

[0097] the ratio R.sub.1 is .ltoreq.0.3, for the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 1, and

[0098] the ratios R.sub.2 and R.sub.3 are .gtoreq.3.0, for the genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 2 and 3, then said biological sample is representative of an acute myeloid leukemia.

[0099] In an embodiment of the invention, when the ratio R.sub.1 is .ltoreq.0.3 and the ratios R.sub.2 and R.sub.3 are both .gtoreq.3, the sample originating from the patient is representative of an acute myeloid leukemia (AML). Said 3 criterions are cumulative.

[0100] AML are clonal proliferation of immature cells of the myeloid origin. They may appear de novo or secondary in patients with myelodysplastic syndrome (MDS). The classification prepared by the French-American-British group (FAB) considers eight varieties (M0-M7) based on morphological criteria and on the immunophenotype of the neoplastic cells (Bennett J M, et al, 1976).

[0101] The World Health Organisation (WHO) classifies AML by incorporating morphological, immunophenotypical, genetic and clinical data to be able to define biological homogeneous entities and with clinical relevance. Thus, AML is classified into four large categories:

1.--AML with recurrent genetic anomalies, 2.--AML with multilineage dysplasia, 3.--AML related to treatment and 4.--non-classifiable AML.

[0102] Before the invention, the cytogenetic analysis represented the most powerful prognosis factor. It is used to identify subgroups of AML with different prognosis: low risk with favourable response to treatment (t(8;21), t(15;17) or inv(16)), intermediate risk (normal karyotype or t(9;11) or high risk (inv(3), del(5q) or del(7q), or more than three alterations). There is molecular heterogeneity within the risk group. In some cases of patients with normal karyotype, the presence of mutations has been found in some genes.

[0103] Advantageously, the invention relates to the method as defined above, wherein if the ratio established in step b). is

[0104] .ltoreq.0.3, for the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 1, and

[0105] .gtoreq.3.0, for the genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 2 and 3, and further wherein the ratio R10 between the expression level of the gene consisting of SEQ ID NO: 10 measured in said sample and in said control sample is lower than 0.5, (R10.ltoreq.0.5), preferably is lower than 0.3, (R10.ltoreq.0.3) then said biological sample is representative of an acute myeloid leukemia.

[0106] In another embodiment, the invention relates to a method as defined above, wherein step c.) is such that

if the ratio established in step b). for each genes of any combination of at least 3 genes from said sub-group is either .gtoreq.2 or .ltoreq.0.5, provided that

[0107] the ratio between the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 1 measured in said biological sample and measured in said control sample, is not .ltoreq.0.3, or

[0108] the ratios between the expression level of each of genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 2 or 3 measured in said biological sample and measured in said control sample, is not .gtoreq.3, then said biological sample is representative of a myelodysplasic disorder, in particular myelodysplasia chosen among refractory anemia (RA), refractory anemia with ringed sideroblasts (RARS), refractory cytopenia with multilineage dysplasia (RCMD), refractory anemia with excess of blasts (RAEB), 5q-syndrome and myelodysplasia unclassifiable.

[0109] In other words, in another advantageous embodiment, the invention relates to a method as defined above, wherein

if the ratio established in step b). for each genes of any combination of at least 3 genes from said sub-group is either .gtoreq.2 or .ltoreq.0.5, provided that if the combination of at least 3 genes corresponds to the genes comprising or being constituted by the nucleic acid sequence SEQ ID NO: 1, 2 and 3, wherein

[0110] the ratio RI is .ltoreq.0.3, for the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 1, and

[0111] the ratios RI are .gtoreq.3.0, for the genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 2 and 3, this combination is excluded then said biological sample is representative of a myelodysplasic disorder, in particular myelodysplasia chosen among refractory anemia with ringed sideroblasts (RARS), refractory cytopenia with multilineage dysplasia (RCMD), refractory anemia with excess of blasts (RAEB) or 5q-syndrome and unclassifiable myelodysplasia.

[0112] According to the invention, if the ratio of at least 3 genes belonging to the sub-group constituted by the genes comprising or being constituted by the nucleic acid SEQ ID NO: 1-6 is either .gtoreq.2 or .ltoreq.0.5, excluding the particular combination of SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 2 wherein the ratio of SEQ ID NO: 1 is .ltoreq.0.3 and the ratios of SEQ ID NO: 2 and 3 are .gtoreq.3, then biological sample is representative of a myelodysplasic disorder, in particular myelodysplasia chosen among refractory anemia with ringed sideroblasts (RARS), refractory cytopenia with multilineage dysplasia (RCMD), refractory anemia with excess of blasts (RAEB) or 5q-syndrome and unclassifiable myelodysplasia.

[0113] According to the invention, myelodysplasic disorders are defined as preleukemia, and correspond to diverse collection of hematological conditions united by ineffective production (or dysplasia) of myeloid blood cells and risk of transformation to acute myelogenous leukemia (AML).

[0114] The French-American-British (FAB) classification has subdivided the myelodysplasic disorders as follows:

Refractory anemia (RA), characterized by less than 5% primitive blood cells (myeloblasts) in the bone marrow and pathological abnormalities primarily seen in red cell precursors, Refractory anemia with ringed sideroblasts (RARS), also characterized by less than 5% myeloblasts in the bone marrow, but distinguished by the presence of 15% or greater red cell precursors in the marrow being abnormal iron-stuffed cells called "ringed sideroblasts", Refractory anemia with excess blasts (RAEB), characterized by 5-20% myeloblasts in the marrow, Refractory anemia with excess blasts in transformation (RAEB-T), characterized by 21-30% myeloblasts in the marrow (>30% blasts is defined as acute myeloid leukemia), and Chronic myelomonocytic leukemia (CMML), not to be confused with chronic myelogenous leukemia or CML, characterized by less than 20% myeloblasts in the bone marrow and greater than 1000.times.10.sup.9/.mu.L monocytes (a type of white blood cell) circulating in the peripheral blood.

[0115] More recently, the World Health Organization (WHO) has classified dysplastic syndromes as follows:

TABLE-US-00002 TABLE 2 Old system New system Refractory Refractory cytopenia with anemia (RA) unilineage dysplasia (Refractory anemia, Refractory neutropenia, and Refractory thrombocytopenia) Refractory Refractory anemia with ring anemia with sideroblasts (RARS) ringed Refractory anemia with ring sideroblasts sideroblasts - thrombocytosis (RARS) (RARS-t) (provisional entity) which is in essence a myelodysplastic/myeloproliferative disorder and usually has a JAK2 mutation (janus kinase) - New WHO classification 2008 Refractory Refractory cytopenia with cytopenia with multilineage dysplasia (RCMD) multilineage includes the subset Refractory dysplasia cytopenia with multilineage (RCMD) dysplasia and ring sideroblasts (RCMD-RS). RCMD includes patients with pathological changes not restricted to red cells (i.e., prominent white cell precursor and platelet precursor (megakaryocyte) dysplasia. Refractory Refractory anemia with excess anemia with blasts I and II. RAEB was divided excess blasts into RAEB-I (5-9% blasts) and (RAEB) RAEB-II (10-19%) blasts, which has a poorer prognosis than RAEB-I. Auer rods may be seen in RAEB-II which may be difficult to distinguish from acute myeloid leukemia. Refractory The category of RAEB-T was anemia with eliminated; such patients are now excess blasts in considered to have acute leukemia. transformation 5q-syndrome, typically seen in (RAEB-T) older women with normal or high platelet counts and isolated deletions of the long arm of chromosome 5 in bone marrow cells, was added to the classification. Chronic CMML was removed from the myelomonocytic myelodysplastic syndromes and put leukemia in a new category of (CMML) myelodysplastic-myeloproliferative overlap syndromes. 5q-syndrome Unclassifiable myelodysplasia (seen in those cases of megakaryocyte dysplasia with fibrosis and others) Refractory cytopenia of childhood (dysplasia in childhood) - New WHO classification 2008

[0116] Chromosome 5q deletion syndrome (chromosome 5q monosomy, 5q-syndrome) is a rare disorder caused by loss of part of the long arm (q arm) of human chromosome 5.

[0117] The 5q-syndrome is characterized by macrocytic anemia and often thrombocytosis, erythroblastopenia, megakaryocyte hyperplasia with nuclear hypolobation and an isolated interstitial deletion of chromosome 5. The 5q-syndrome is found predominantly in females of advanced age.

[0118] In still another embodiment, the invention relates to a method above defined, wherein

said set comprises 10 genes, said 10 genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 1 to 10, and the step c). is such that

[0119] if the ratio established in step b). for each genes of any combination of 3 genes from said sub-group is either .gtoreq.2 or .ltoreq.0.5, provided that

[0120] the ratio between the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 1 measured in said biological sample and measured in said control sample, is not .ltoreq.0.3, or

[0121] the ratios between the expression level of each of genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 2 or 3 measured in said biological sample and measured in said control sample, is not .gtoreq.3 and further

[0122] if the ratio established in step b) for at least one gene of said set that does not belong to said subgroup is .gtoreq.2 and the ratio established in step b) of at least one other gene of said set that does not belong to said subgroup is .ltoreq.0.5, then said biological sample is representative of a refractory anemia with excess of blast or of a 5q-syndrome.

[0123] In other words, in still another advantageous embodiment, the invention relates to a method above defined, wherein

said set comprises 10 genes, said 10 genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 1 to 10, and the step c). is such that

[0124] if the ratio established in step b). for each genes of any combination of 3 genes from said sub-group is either .gtoreq.2 or .ltoreq.0.5, provided that if the combination of at least 3 genes corresponds to the genes comprising or being constituted by the nucleic acid sequence SEQ ID NO: 1, 2 and 3, wherein

[0125] the ratio RI is .ltoreq.0.3, for the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 1, and

[0126] the ratios RI are .gtoreq.3.0, for the genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 2 and 3, this combination is excluded, and further

[0127] if the ratio established in step b) for at least one gene of said set that does not belong to said subgroup is .gtoreq.2 and the ratio established in step b) of at least one other gene of said set that does not belong to said subgroup is .ltoreq.0.5, then said biological sample is representative of a refractory anaemia with excess of blast or of a 5q-syndrome.

[0128] In this advantageous embodiment of the invention, the genes belonging to the set (i.e. genes comprising or being constituted by SEQ ID NO: 1 to 10) are helpful for discriminating the myelodysplastic disorders.

[0129] More precisely, the genes belonging to the set but that do not belong to the sub group (i.e. genes comprising or being constituted by SEQ ID NO: 7 to 10-D in FIG. 2) are helpful for discriminating the myelodysplastics disorders.

[0130] Then, if, by measuring the expression level of the genes SEQ ID NO: 1-6, the biological sample is considered to be representative of myelodyplastic disorder, it is possible according to the invention to separate refractory anemia with excess of blast and of a 5q-syndrome from the other pathologies, when the ratio of the expression of at least one gene of the group consisting of SEQ ID NO: 7 to 10 is .gtoreq.2 and when the ratio of the expression of at least one gene of the group consisting of SEQ ID NO: 7 to 10 is .ltoreq.0.5.

[0131] Another advantageous embodiment of the invention relates to a method previously defined, wherein

said set comprises 10 genes, said 10 genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 1 to 10, and the step c). is such that

[0132] if the ratio established in step b). for each genes of any combination of 3 genes from said sub-group is either .gtoreq.2 or .ltoreq.0.5, provided that

[0133] the ratio between the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 1 measured in said biological sample and measured in said control sample, is not .ltoreq.0.3, or

[0134] the ratios between the expression level of each of genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 2 or 3 measured in said biological sample and measured in said control sample, is not .gtoreq.3, and

[0135] if the ratio established in step b) for at least one gene of said set that does not belong to said subgroup is .gtoreq.2 and the ratio established in step b) of at least one other gene of said set that does not belong to said subgroup is .ltoreq.0.5, and further if the ratio established in step b) for at least 4 genes of the group of 24 genes that does not belong to said set is .gtoreq.3, then said biological sample is representative of a refractory anemia with excess of blast.

[0136] In other words, another advantageous embodiment of the invention relates to a method previously defined, wherein

said set comprises 10 genes, said 10 genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 1 to 10, and the step c). is such that

[0137] if the ratio established in step b). for each genes of any combination of 3 genes from said sub-group is either .gtoreq.2 or .ltoreq.0.5, provided that if the combination of at least 3 genes corresponds to the genes comprising or being constituted by the nucleic acid sequence SEQ ID NO: 1, 2 and 3, wherein

[0138] the ratio R.sub.i is .ltoreq.0.3, for the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 1, and

[0139] the ratios R.sub.i are .gtoreq.3.0, for the genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 2 and 3, this combination is excluded, and

[0140] if the ratio established in step b) for at least one gene of said set that does not belong to said subgroup is .gtoreq.2 and the ratio established in step b) of at least one other gene of said set that does not belong to said subgroup is .ltoreq.0.5, and further

[0141] if the ratio established in step b) for at least 4 genes of the group of 24 genes that does not belong to said set is .gtoreq.3, then said biological sample is representative of a refractory anaemia with excess of blast.

[0142] In this advantageous embodiment of the invention, the genes belonging to the group (i.e. genes comprising or being constituted by SEQ ID NO: 1 to 24) are helpful for discriminating between refractory anaemia with excess of blast and of a 5q-syndrome.

[0143] More precisely, the genes belonging to the group but that do not belong to the set (i.e. genes comprising or being constituted by SEQ ID NO: 11 to 24-E in FIG. 2) are helpful for the discrimination between refractory anaemia with excess of blast and of a 5q-syndrome.

[0144] The invention also relates to a method for the diagnosis, and/or the classification, preferably in vitro, of an hematological disorder, in particular myeloid and/or lymphoid hematological disorder, preferably myeloid hematological disorder,

said method comprising the steps of: a). measuring, from cells contained in a biological sample of a subject, preferably from blood cells or bone marrow cells containing sample, the expression level of at least the genes of a sub-group of 6 genes belonging to a set of genes chosen among a group of 24 genes, said group of 24 genes comprising or being constituted by the nucleic acid sequences SEQ ID NO:1 to 24, said 6 genes belonging to said sub-group comprising or being constituted by the nucleic acid sequences SEQ ID NO: 1 to 6, b). comparing the expression level of each genes measured in step a)., with the expression level of the same respective genes from cells contained in a control sample, preferably from a control sample containing blood cells or bone marrow cells, said control sample being of the same nature than said biological sample, to establish a gene expression level ratio Ri between the expression level of each genes measured in step a) and the expression level of the same respective genes from cells contained in a control sample, for each genes of said sub-group, and c). determining the status of said biological sample such that if the ratio Ri for each genes of any combination of 3 genes from said sub-group is

[0145] either .gtoreq.2,

[0146] or .ltoreq.0.5,

[0147] said biological sample is representative of an hematological disorder cells.

[0148] An advantageous embodiment of the invention relates to a method as defined above, wherein if

[0149] the ratio R.sub.i is .ltoreq.0.3, for the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 1, and

[0150] the ratios R.sub.i are .gtoreq.3.0, for the genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 2 and 3, then said biological sample is representative of an acute myeloid leukemia.

[0151] In this advantageous embodiment of the method according to the invention, in step c). when the ratio Ri of the expression level of the genes GPX3 (SEQ ID NO: 1) .ltoreq.0.3, the ratio of the expression level of the genes GPX1(1) (SEQ ID NO: 2) is .gtoreq.3.0 and the ratio of the expression level of the genes GLRX2(2) (SEQ ID NO: 3) is .gtoreq.3.0, then the biological sample is representative of an acute myeloid leukemia (AML).

[0152] In another advantageous embodiment, the invention relates to a method as defined above, wherein step c.) is such that

if the ratio R.sub.i established in step b). for each genes of any combination of at least 3 genes from said sub-group is either .gtoreq.2 or .ltoreq.0.5, provided that if the combination of 3 genes corresponds to the genes comprising or being constituted by the nucleic acid sequence SEQ ID NO: 1, 2 and 3, wherein

[0153] the ratio R.sub.i is .ltoreq.0.3, for the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 1, and

[0154] the ratios R.sub.i are .gtoreq.3.0, for the genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 2 and 3, said combination is excluded then said biological sample is representative of a myelodysplasic disorder, in particular myelodysplasia chosen among refractory anemia with ringed sideroblasts (RARS), refractory cytopenia with multilineage dysplasia (RCMD), refractory anemia with excess of blasts (RAEB) or 5q-syndrome and unclassifiable myelodysplasia.

[0155] In other words, when the ratio Ri, established in step c)., of the expression level of each gene of any combination of at least 3 genes chosen among the genes comprising or being constituted by SEQ ID NO: 1-6 is either .gtoreq.2 or .ltoreq.0.5, and said combination does not corresponds to the combination that defines a biological sample as representative of an AML, then said sample is representative of a myelodysplastic disorder or syndrome.

[0156] Another advantageous embodiment of the invention relates to a method previously defines, wherein

said set comprises 10 genes, said 10 genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 1 to 10, and further, in step c.) if the ratio Ri of all the genes of said set that do not belong to said subgroup is comprised between 0.3 to 2, the extremity of the interval being excluded then said biological sample is representative of a refractory anemia with ringed sideroblasts or a refractory cytopenia with multilineage dysplasia.

[0157] In the invention "the genes of said set that do not belong to said subgroup" corresponds to the genes belonging to the group D as defined in FIG. 2.

[0158] Since the subgroup consists of the genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 1-6 and the set consists of the genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 1-10, consequently, the genes of said set that do not belong to said subgroup correspond to the genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 7-10.

[0159] In the above embodiment, if the ratio R.sub.i for each gene represented by the nucleic acid sequences SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9 and SEQ ID NO: 10, i.e. all the ratios of said genes, is comprised between 0.3 and 2, 0.3 and 2 being excluded from the interval, said biological sample is representative of a refractory anemia with ringed sideroblasts or a refractory cytopenia with multilineage dysplasia.

[0160] It is possible to write that if 0.3<R7<2, and 0.3<R8<2, and 0.3<R9<2, and 0.3<R10<2, R7, R8, R9 and R10 representing the respective ratio for the genes represented by SEQ ID NO: 7, 8, 9 and 10), further to the evaluation of the ratio of the genes SEQ ID NO: 1-6, then said biological sample is representative of a refractory anemia with ringed sideroblasts or a refractory cytopenia with multilineage dysplasia.

[0161] The terms "comprised between 0.3 to 2, the extremity of the interval being excluded" refer to the interval represented by the mathematical symbol: ]0.3;2.0[. This interval includes all the values comprised between 0.3 and 2.0, but excludes the specific values 0.3 and 2.

[0162] If Ri=0.3, or Ri=2.0, Ri does not belong to the interval ]0.3;2.0[.

[0163] The evaluation of the ratio of the expression level of the genes represented by SEQ ID NO: 7-10, in a specific interval, allows to discriminate some myelodysplastic syndrome.

[0164] To summarise,

if the ratio for each gene of any combination of 3 genes chosen among the 6 genes represented by SEQ ID NO: 1, 2, 3, 4, 5 and 6, is either .gtoreq.2.0 or .ltoreq.0.5, excluding the combination that defines leukemia, and the ratio of each gene represented by SEQ ID NO: 7, 8, 9 and 10 is comprised in the interval ]0.3; 2.0[, then the biological sample is representative of a refractory anemia with ringed sideroblasts or a refractory cytopenia with multilineage dysplasia.

[0165] In one another advantageous embodiment, the invention relates to a method according the definition mentioned above, wherein further, in step c.)

[0166] if the ratio Ri of at least one gene belonging to said group of genes that does not belong to said set is .ltoreq.0.3, then said biological sample is representative of a refractory anemia with ringed sideroblasts, and

[0167] if the ratio RI of at least one gene belonging to said group of genes that do not belong to said set is .gtoreq.3.0,

[0168] then said biological sample is representative of a refractory cytopenia with multilineage dysplasia.

[0169] In the invention "gene belonging to said group of genes that does not belong to said set" corresponds to the genes belong ing to the group E as defined in FIG. 2.

[0170] Since the subgroup consists of the genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 1-6, the set consists of the genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 1-10, and the group consists of the genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 1-24, consequently, the genes of said group that do not belong to said set correspond to the genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 11-24.

[0171] In the above embodiment, the genes represented by SEQ ID NO: 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 and 24 provide supplemental information regarding the nature of the tested biological sample.

[0172] To summarise,

1) a) if the ratio for each gene of any combination of 3 genes chosen among the 6 genes represented by SEQ ID NO: 1, 2, 3, 4, 5 and 6, is either .gtoreq.2.0 or .ltoreq.0.5, excluding the combination that defines leukemia, and b) the ratio of each gene represented by SEQ ID NO: 7, 8, 9 and 10 is comprised in the interval ]0.3; 2.0[, and c) the ratio of at least 1 gene, i.e. 1, or 2, or 3, or 4, or 5, or 6, or 7, or 8, or 9, or 10, or 11, or 12, or 13, or 14 genes, represented by SEQ ID NO: 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 and 24 is .gtoreq.3.0, then said biological sample is representative of a refractory cytopenia with multilineage dysplasia, and 2)) a) if the ratio for each gene of any combination of 3 genes chosen among the 6 genes represented by SEQ ID NO: 1, 2, 3, 4, 5 and 6, is either .gtoreq.2.0 or .ltoreq.0.5, excluding the combination that defines leukemia, and b) the ratio of each gene represented by SEQ ID NO: 7, 8, 9 and 10 is comprised in the interval ]0.3; 2.0[, and c) the ratio of at least 1 gene, i.e. 1, or 2, or 3, or 4, or 5, or 6, or 7, or 8, or 9, or 10, or 11, or 12, or 13, or 14 genes, represented by SEQ ID NO: 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 and 24 is .ltoreq.0.3, then said biological sample is representative of a refractory anemia with ringed sideroblasts.

[0173] In one other embodiment, the invention relates to a method as defined above, wherein

said set comprises 10 genes, said 10 genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 1 to 10, and further if

[0174] the ratio R.sub.i of at least one gene of said set that does not belong to said subgroup is .gtoreq.2.0, or

[0175] the ratio R.sub.i of at least one gene of said set that does not belong to said subgroup is .ltoreq.0.3, or

[0176] the ratio R.sub.i of at least one gene of said set that does not belong to said subgroup is .gtoreq.2.0 and the ratio Ri of another gene that does not belong to said subgroup is .ltoreq.0.3,

[0177] then said biological sample is representative of chosen among refractory cytopenia with multilineage dysplasia, refractory anemia with excess of blasts, 5 q-syndroma and unclassified myelodysplasia.

[0178] In this advantageous embodiment, it is taking account of the case wherein at least one gene represented by SEQ ID NO: 7, 8 9 or 10 does not belong to the interval ]0.3; 2.0[ as defined above.

[0179] In a first case, at least one gene has a ratio .gtoreq.2.0, whatsoever the ratio of the other genes. In a second case at least one gene has a ratio .ltoreq.0.3, whatsoever the ratio of the other genes. In a third case, both at least one gene has a ratio .gtoreq.2.0 and another gene has a ratio .ltoreq.0.3, whatsoever the ratio of the other genes.

[0180] In one other advantageous embodiment, the invention relates to a method previously defined, wherein

if the ratio R.sub.i of at least one gene of said set that does not belong to said subgroup is .gtoreq.2.0 and the ratio R.sub.i of at least another gene of said set that does not belong to said subgroup is .ltoreq.0.5, then said biological sample is a representative refractory anemia with excess of blasts or of 5q-syndrome.

[0181] This embodiment concerns the case in which at least one gene has a ratio .gtoreq.2.0, whatsoever the ratio of the other genes, and in particular the case in which at least one gene has a ratio .gtoreq.2.0 and another gene has a ratio .ltoreq.0.5, and therefore possibly .ltoreq.0.3.

[0182] This particular situation allows to detect, or to identify, myelodysplastic syndromes that are representative refractory anemia with excess of blasts or of 5q-syndrome.

[0183] In one other advantageous embodiment, the invention relates to a method previously defined, wherein

if the ratio R.sub.i of at least one gene of said set that does not belong to said subgroup is .gtoreq.2.0 and the ratio R.sub.i of at least another gene of said set that does not belong to said subgroup is .ltoreq.0.5, then said biological sample is a representative refractory anemia with excess of blast or 5q-syndrome, and further

[0184] if the ratios R.sub.i of at least four genes belonging to said group of genes that do not belong to said set are .gtoreq.3.0, then said biological sample is representative of a refractory anemia with excess of blasts, and

[0185] if the ratios R.sub.i of at most three genes belonging to said group of genes that do not belong to said set are .gtoreq.3.0, then said biological sample is representative of a 5q-syndrome.

[0186] To summarise,

1) a) if the ratio for each gene of any combination of 3 genes chosen among the 6 genes represented by SEQ ID NO: 1, 2, 3, 4, 5 and 6, is either .gtoreq.2.0 or .ltoreq.0.5, excluding the combination that defines leukemia, and b) the ratio of at least one gene represented by SEQ ID NO: 7, 8, 9 and 10 is .gtoreq.2.0, and at least one other gene represented by SEQ ID NO: 7, 8, 9 and 10 is .ltoreq.0.5, and c) the ratios of at least 4 genes, i.e. 4, or 5, or 6, or 7, or 8, or 9, or 10, or 11, or 12, or 13, or 14 genes, represented by SEQ ID NO: 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 and 24 are .gtoreq.3.0, then said biological sample is representative of refractory anemia with excess of blasts, and 2)) a) if the ratio for each gene of any combination of 3 genes chosen among the 6 genes represented by SEQ ID NO: 1, 2, 3, 4, 5 and 6, is either .gtoreq.2.0 or .ltoreq.0.5, excluding the combination that defines leukemia, and b) the ratio of at least one gene represented by SEQ ID NO: 7, 8, 9 and 10 is .gtoreq.2.0, and at least one other gene represented by SEQ ID NO: 7, 8, 9 and 10 is .ltoreq.0.5, and c) the ratio of at most 3 genes, i.e. 0, or 1, or 2, or 3, genes, represented by SEQ ID NO: 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 and 24 are .gtoreq.3.0, then said biological sample is representative of a 5q-syndrome.

[0187] In one other advantageous embodiment, the invention relates to a method according the above definition,

wherein if

[0188] the ratio R.sub.i of at least one gene of said set that does not belong to said subgroup is .gtoreq.2.0 and

[0189] the ratio R.sub.i of no gene of said set that does not belong to said subgroup is .ltoreq.0.5,

[0190] then said biological sample is representative of a refractory cytopenia with multilineage dysplasia,

[0191] In this specific embodiment, if the ratio of at least one gene represented by SEQ ID NO: 7, 8 9 or 10 is .gtoreq.2.0 and the ratio of the other genes is included in the interval [ 0.5; +.infin.[, then said biological sample is representative of a refractory cytopenia with multi lineage dysplasia.

[0192] To summarize:

a) if the ratio for each gene of any combination of 3 genes chosen among the 6 genes represented by SEQ ID NO: 1, 2, 3, 4, 5 and 6, is either .gtoreq.2.0 or .ltoreq.0.5, excluding the combination that defines leukemia, and b) the ratio of at least one gene represented by SEQ ID NO: 7, 8, 9 and 10 is .gtoreq.2.0, and the ratio of no gene represented by SEQ ID NO: 7, 8, 9 and 10 is .ltoreq.0.5, then said biological sample is representative of a refractory cytopenia with multilineage dysplasia.

[0193] In one other advantageous embodiment, the invention relates to a method according the above definition,

wherein further if

[0194] the ratio R.sub.i of at least one gene of said set that does not belong to said subgroup is .ltoreq.0.5 and

[0195] the ratio RI of no gene of said set that does not belong to said subgroup is .gtoreq.2.0,

[0196] then said biological sample is representative of refractory anemia with ringed sideroblasts, refractory cytopenia with multilineage dysplasia, refractory anemia with excess of blasts, 5q-syndrome or unclassified myelodysplasia,

[0197] In this specific embodiment, if the ratio of at least one gene represented by SEQ ID NO: 7, 8 9 or 10 is .ltoreq.0.5 and the ratio of the other genes is included in the interval ]-.infin.; 2.0], then said biological sample is representative of refractory anemia with ringed sideroblasts, refractory cytopenia with multilineage dysplasia, refractory anemia with excess of blasts, 5q-syndrome or unclassified myelodysplasia.

[0198] To summarize:

a) if the ratio for each gene of any combination of 3 genes chosen among the 6 genes represented by SEQ ID NO: 1, 2, 3, 4, 5 and 6, is either .gtoreq.2.0 or .ltoreq.0.5, excluding the combination that defines leukemia, and b) the ratio of at least one gene represented by SEQ ID NO: 7, 8, 9 and 10 is .ltoreq.0.5, and the ratio of no gene represented by SEQ ID NO: 7, 8, 9 and 10 is .gtoreq.2.0, then said biological sample is representative of refractory anemia with ringed sideroblasts, refractory cytopenia with multilineage dysplasia, refractory anemia with excess of blasts, 5q-syndrome or unclassified myelodysplasia.

[0199] In one other advantageous embodiment, the invention relates to a method as defined above, wherein, in step c.)

if

[0200] the ratio R.sub.i of at least one gene of said set that does not belong to said subgroup is .ltoreq.0.5 and

[0201] the ratio R.sub.i of no gene of said set that does not belong to said subgroup is .gtoreq.2.0,

[0202] and further

[0203] if

[0204] the ratios R.sub.i of all the genes belonging to said group of genes that do not belong to said set are comprised between 0.5 to 2, the extremity of the interval being excluded,

[0205] then said biological sample is representative of a unclassified myelodysplasia,

[0206] In this advantageous embodiment the genes represented by SEQ ID NO: 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 are useful for discriminate unclassified myelodysplasia from refractory anemia with ringed sideroblasts, refractory cytopenia with multilineage dysplasia and 5q-syndrome.

[0207] In one other advantageous embodiment, the invention relates to a method as defined above, wherein, in step c.)

if

[0208] the ratio R.sub.i of at least one gene of said set that does not belong to said subgroup is .ltoreq.0.3 and

[0209] the ratio R.sub.i of no gene of said set that does not belong to said subgroup is .gtoreq.2.0,

[0210] and further

[0211] if

[0212] the ratios R.sub.i of at least one gene belonging to said group of genes that do not belong to said set is .gtoreq.2.0, then said biological sample is representative of a 5q-syndrome or refractory cytopenia with multilineage dysplasia.

[0213] To summarize:

a) if the ratio for each gene of any combination of 3 genes chosen among the 6 genes represented by SEQ ID NO: 1, 2, 3, 4, 5 and 6, is either .gtoreq.2.0 or .ltoreq.0.5, excluding the combination that defines leukemia, and b) the ratio of at least one gene represented by SEQ ID NO: 7, 8, 9 and 10 is .ltoreq.0.3, and the ratio of no gene represented by SEQ ID NO: 7, 8, 9 and 10 is .gtoreq.2.0, and c) the ratios RI of at least one gene represented by SEQ ID NO: 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 is .gtoreq.2.0, then said biological sample is representative of a 5q-syndrome or refractory cytopenia with multilineage dysplasia.

[0214] Another advantageous embodiment of the invention relates to a method as defined above, wherein

if the ratio R.sub.i of at least one gene of said set that does not belong to said subgroup is .ltoreq.0.3 and the ratio R.sub.i of no gene that does not belong to said subgroup is .gtoreq.2.0, then said biological sample is representative of a unclassified myelodysplasia, a 5q-syndrome, or of a refractory cytopenia with multilineage dysplasia.

[0215] To summarize:

a) if the ratio for each gene of any combination of 3 genes chosen among the 6 genes represented by SEQ ID NO: 1, 2, 3, 4, 5 and 6, is either .gtoreq.2.0 or .ltoreq.0.5, excluding the combination that defines leukemia, and b) the ratio of at least one gene represented by SEQ ID NO: 7, 8, 9 and 10 is .ltoreq.0.3, and the ratio of no gene represented by SEQ ID NO: 7, 8, 9 and 10 is .gtoreq.2.0, said biological sample is representative of a unclassified myelodysplasia, a 5q-syndrome, or of a refractory cytopenia with multilineage dysplasia.

[0216] Another advantageous embodiment of the invention relates to a method as defined above, wherein

if the ratio R.sub.i of at least one gene of said set that does not belong to said subgroup is .ltoreq.0.3 and the ratio R.sub.i of no gene that does not belong to said subgroup is .gtoreq.2.0, and further if the ratios R.sub.i of at least two genes of said set that do not belong to said subgroup are .ltoreq.0.3 and the ratio R.sub.i of no gene of said set that does not belong to said subgroup is .gtoreq.2.0, then said biological sample is representative of a 5q-syndrome, or of refractory cytopenia with multilineage dysplasia.

[0217] To summarize:

a) if the ratio for each gene of any combination of 3 genes chosen among the 6 genes represented by SEQ ID NO: 1, 2, 3, 4, 5 and 6, is either .gtoreq.2.0 or .ltoreq.0.5, excluding the combination that defines leukemia, and b) the ratio of at least one gene represented by SEQ ID NO: 7, 8, 9 and 10 is .ltoreq.0.3, the ratio of at least one other gene represented by SEQ ID NO: 7, 8, 9 and 10 is .ltoreq.0.3 and the ratio of no gene represented by SEQ ID NO: 7, 8, 9 and 10 is .gtoreq.2.0, said biological sample is representative of a 5q-syndrome, or of a refractory cytopenia with multilineage dysplasia.

[0218] Another advantageous embodiment of the invention relates to a method as defined above, wherein

if the ratio R.sub.i of at least one gene of said set that does not belong to said subgroup is .ltoreq.0.3 and the ratio R.sub.i of no gene that does not belong to said subgroup is .gtoreq.2.0, and if the ratios R.sub.i of at least two genes of said set that do not belong to said subgroup are .ltoreq.0.3 and the ratio R.sub.i of no gene of said set that does not belong to said subgroup is .gtoreq.2.0, then said biological sample is representative of a 5q-syndrome, or of refractory cytopenia with multilineage dysplasia. and

[0219] if further the ratios R.sub.i of at least two genes belonging to said group of genes that do not belong to said set are .gtoreq.2.0, then said biological sample is representative of a 5q-syndrome.

[0220] To summarize:

a) if the ratio for each gene of any combination of 3 genes chosen among the 6 genes represented by SEQ ID NO: 1, 2, 3, 4, 5 and 6, is either .gtoreq.2.0 or .ltoreq.0.5, excluding the combination that defines leukemia, and b) the ratio of at least one gene represented by SEQ ID NO: 7, 8, 9 and 10 is .ltoreq.0.3, the ratio of at least one other gene represented by SEQ ID NO: 7, 8, 9 and 10 is .ltoreq.0.3 and the ratio of no gene represented by SEQ ID NO: 7, 8, 9 and 10 is .gtoreq.2.0, and c) the ratios of at least two genes represented by SEQ ID NO: 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 are .gtoreq.2.0, said biological sample is representative of a 5q-syndrome.

[0221] The invention also relates to a method for the diagnosis, and/or the classification, preferably in vitro, of an hematological disorder, in particular myeloid and/or lymphoid hematological disorder, preferably myeloid hematological disorder,

said method comprising the steps of: a). measuring, from cells contained in a biological sample of a subject, preferably from blood cells or bone marrow cells containing sample, the expression level 24 genes, said 24 genes comprising or being constituted by the nucleic acid sequences SEQ ID NO:1 to 24, b). comparing the expression level of each genes measured in step a)., with the expression level of the same respective genes from cells contained in a control sample preferably from blood cells or bone marrow cells containing sample, said control sample being of the same nature than said biological sample, to establish a gene expression level ratio for each genes of said sub-group, and c). determining the status of said biological sample such that if the ratio established in step b).

[0222] More advantageously, the invention relates to a method previously defined, wherein the expression level of the genes is measured by a method allowing the determination of the amount of the mRNA or of the cDNA corresponding to said genes. Preferably said method is a quantitative method.

[0223] Levels of mRNA can be quantitatively measured by northern blotting which gives size and sequence information about the mRNA molecules. A sample of RNA is separated on an agarose gel and hybridized to a radio-labeled RNA probe that is complementary to the target sequence. The radio-labeled RNA is then detected by an autoradiograph. Northern blotting is widely used as the additional mRNA size information allows the discrimination of alternately spliced transcripts.

[0224] Another approach for measuring mRNA abundance is reverse transcription quantitative polymerase chain reaction (RT-PCR followed with qPCR). RT-PCR first generates a DNA template from the mRNA by reverse transcription, which is called cDNA. This cDNA template is then used for qPCR where the change in fluorescence of a probe changes as the DNA amplification process progresses. With a carefully constructed standard curve qPCR can produce an absolute measurement such as number of copies of mRNA, typically in units of copies per nanolitre of homogenized tissue or copies per cell. qPCR is very sensitive (detection of a single mRNA molecule is possible), but can be expensive due to the fluorescent probes required.

[0225] Northern blots and RT-qPCR are good for detecting whether a single gene or few genes are expressed.

[0226] Other methods known for one skilled in the art include DNA microarrays or technologies like Serial Analysis of Gene Expression (SAGE).

[0227] SAGE can provide a relative measure of the cellular concentration of different messenger RNAs. The great advantage of tag-based methods is the "open architecture", allowing for the exact measurement of any transcript are present in cells, the sequence of said transcripts could be known or unknown.

[0228] The preferred method used according to the invention is RT-qPCR.

[0229] In still another advantageous embodiment, the invention relates to a method above defined, wherein the measure of the expression level of the genes is carried out by using the at least 6 pairs of oligonucleotides belonging to a group of 24 pairs of oligonucleotides comprising or being constituted by the nucleic acid sequences SEQ ID NO: 25 to 72, said at least 6 pairs of oligonucleotides comprising or being constituted by the nucleic acid sequences SEQ ID NO: 25 to 36.

[0230] In the invention:

[0231] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 25 and SEQ ID NO: 26, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 1,

[0232] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 27 and SEQ ID NO: 28, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 2,

[0233] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 29 and SEQ ID NO: 30, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 3,

[0234] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 31 and SEQ ID NO: 32, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 4,

[0235] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 33 and SEQ ID NO: 34, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 5,

[0236] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 35 and SEQ ID NO: 36, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 6.

[0237] In one advantageous embodiment, the inventions relates to the method as defined above,

[0238] using the at least 10 pairs of oligonucleotides belonging to a group of 24 pairs of oligonucleotides comprising or being constituted by the nucleic acid sequences SEQ ID NO: 25 to 72, said at least 6 pairs of oligonucleotides comprising or being constituted by the nucleic acid sequences SEQ ID NO: 25 to 44.

[0239] In the invention:

[0240] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 36 and SEQ ID NO: 37, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 7,

[0241] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 38 and SEQ ID NO: 39, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 8,

[0242] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 41 and SEQ ID NO: 42, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 9,

[0243] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 43 and SEQ ID NO: 44, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 10.

[0244] In one advantageous embodiment, the inventions relates to the method as defined above,

using 24 pairs of oligonucleotides comprising or being constituted by the nucleic acid sequences SEQ ID NO: 25 to 72.

[0245] In the invention:

[0246] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 45 and SEQ ID NO: 46, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 11,

[0247] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 47 and SEQ ID NO: 48, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 12,

[0248] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 49 and SEQ ID NO: 50, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 13,

[0249] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 51 and SEQ ID NO: 52, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 14,

[0250] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 53 and SEQ ID NO: 54, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 15,

[0251] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 55 and SEQ ID NO: 56, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 16,

[0252] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 57 and SEQ ID NO: 58, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 17,

[0253] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 59 and SEQ ID NO: 60, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 18,

[0254] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 61 and SEQ ID NO: 62, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 19,

[0255] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 63 and SEQ ID NO: 64, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 20,

[0256] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 65 and SEQ ID NO: 66, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 21,

[0257] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 67 and SEQ ID NO: 68, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 22,

[0258] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 69 and SEQ ID NO: 70, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 23,

[0259] the pair of oligonucleotides comprising of being constituted by the nucleic acid sequences SEQ ID NO: 71 and SEQ ID NO: 72, are used for measuring the expression level of the gene comprising or being constituted by the nucleic acid sequence SEQ ID NO: 24.

[0260] The following table 3 represents genes/variants/oligonucleotides used according to the invention.

TABLE-US-00003 TABLE 3 Table 23 SEQ ID SEQ ID Gene Oligo Oligo name SEQ ID SEQ ID variant forward reverse GPX3 SEQ ID -- SEQ ID SEQ ID NO: 1 NO: 25 NO: 26 GPX1 SEQ ID -- SEQ ID SEQ ID (1) NO: 2 NO: 27 NO: 28 GLRX2 SEQ ID -- SEQ ID SEQ ID (2) NO: 3 NO: 29 NO: 30 CAT SEQ ID -- SEQ ID SEQ ID NO: 4 NO: 31 NO: 32 PRDX SEQ ID SEQ ID NO: 73 SEQ ID SEQ ID (1-2-3) NO: 5 or NO: 33 NO: 34 SEQ ID NO: 74 PRDX5 SEQ ID -- SEQ ID SEQ ID (2) NO: 6 NO: 35 NO: 36 SOD2 SEQ ID SEQ ID NO: 75 SEQ ID SEQ ID (1-2-3) NO: 7 or NO: 37 NO: 38 SEQ ID NO: 76 GSR SEQ ID SEQ ID NO: 77 SEQ ID SEQ ID (1-2-3-4) NO: 8 or NO: 39 NO: 40 SEQ ID NO: 78 or SEQ ID NO: 79 GLRX SEQ ID SEQ ID NO: 80 SEQ ID SEQ ID (1-2) NO: 9 NO: 41 NO: 42 PRDX2 SEQ ID -- SEQ ID SEQ ID (1) NO: 10 NO: 43 NO: 44 PRDX5 SEQ ID SEQ ID NO: 81 SEQ ID SEQ ID (1-3) NO: 11 or NO: 45 NO: 46 SEQ ID NO: 82 SOD1 SEQ ID -- SEQ ID SEQ ID NO: 12 NO: 47 NO: 48 TXN SEQ ID -- SEQ ID SEQ ID NO: 13 NO: 49 NO: 50 PRDX3 SEQ ID SEQ ID NO: 83 SEQ ID SEQ ID (1-2) NO: 14 NO: 51 NO: 52 GPX7 SEQ ID -- SEQ ID SEQ ID NO: 15 NO: 53 NO: 54 GPX4 SEQ ID SEQ ID NO: 84 SEQ ID SEQ ID (1-2-3) NO: 16 or NO: 55 NO: 56 SEQ ID NO: 85 TXN2 SEQ ID -- SEQ ID SEQ ID NO: 17 NO: 57 NO: 58 PRDX4 SEQ ID -- SEQ ID SEQ ID NO: 18 NO: 59 NO: 60 GPX1(2) SEQ ID -- SEQ ID SEQ ID NO: 19 NO: 61 NO: 62 GLRX3 SEQ ID -- SEQ ID SEQ ID NO: 20 NO: 63 NO: 64 PRDX2 SEQ ID -- SEQ ID SEQ ID (3) NO: 21 NO: 65 NO: 66 PRDX6 SEQ ID -- SEQ ID SEQ ID NO: 22 NO: 67 NO: 68 GLRX5 SEQ ID -- SEQ ID SEQ ID NO: 23 NO: 69 NO: 70 GLRX2 SEQ ID -- SEQ ID SEQ ID (1) NO: 24 NO: 71 NO: 72 GPX3 SEQ ID -- SEQ ID SEQ ID NO: 1 NO: 25 NO: 26 GPX1 SEQ ID -- SEQ ID SEQ ID (1) NO: 2 NO: 27 NO: 28 GLRX2 SEQ ID -- SEQ ID SEQ ID (2) NO: 3 NO: 29 NO: 30 CAT SEQ ID -- SEQ ID SEQ ID NO: 4 NO: 31 NO: 32 PRDX SEQ ID SEQ ID NO: 73 SEQ ID SEQ ID (1-2-3) NO: 5 or NO: 33 NO: 34 SEQ ID NO: 74 PRDX5 SEQ ID -- SEQ ID SEQ ID (2) NO: 6 NO: 35 NO: 36 SOD2 SEQ ID SEQ ID NO: 75 SEQ ID SEQ ID (1-2-3) NO: 7 or NO: 37 NO: 38 SEQ ID NO: 76

[0261] The oligonucleotides defined above are preferably used for carrying out a qPCR reaction.

[0262] qPCR is well known in the art, and can be carried out by using, in association with oligonucleotides allowing a specific amplification of the target gene, either with dyes or with reporter probe.

[0263] Both techniques are briefly summarized hereafter.

[0264] Real-Time PCR with Double-Stranded DNA-Binding Dyes as Reporters:

[0265] A DNA-binding dye binds to all double-stranded (ds) DNA in PCR, causing fluorescence of the dye. An increase in DNA product during PCR therefore leads to an increase in fluorescence intensity and is measured at each cycle, thus allowing DNA concentrations to be quantified.

[0266] However, dsDNA dyes such as SYBR Green will bind to all dsDNA PCR products, including nonspecific PCR products (such as Primer dimer). This can potentially interfere with or prevent accurate quantification of the intended target sequence.

[0267] The reaction is prepared as usual, with the addition of fluorescent dsDNA dye.

[0268] The reaction is run in a Real-time PCR instrument, and after each cycle, the levels of fluorescence are measured with a detector; the dye only fluoresces when bound to the dsDNA (i.e., the PCR product). With reference to a standard dilution, the dsDNA concentration in the PCR can be determined.

[0269] Like other real-time PCR methods, the values obtained do not have absolute units associated with them (i.e., mRNA copies/cell). As described above, a comparison of a measured DNA/RNA sample to a standard dilution will only give a fraction or ratio of the sample relative to the standard, allowing only relative comparisons between different tissues or experimental conditions. To ensure accuracy in the quantification, it is usually necessary to normalize expression of a target gene to a stably expressed gene (see below). This can correct possible differences in RNA quantity or quality across experimental samples.

[0270] Fluorescent Reporter Probe Method

[0271] Fluorescent reporter probes detect only the DNA containing the probe sequence; therefore, use of the reporter probe significantly increases specificity, and enables quantification even in the presence of non-specific DNA amplification. Fluorescent probes can be used in multiplex assays--for detection of several genes in the same reaction--based on specific probes with different-coloured labels, provided that all targeted genes are amplified with similar efficiency. The specificity of fluorescent reporter probes also prevents interference of measurements caused by primer dimers, which are undesirable potential by-products in PCR. However, fluorescent reporter probes do not prevent the inhibitory effect of the primer dimers, which may depress accumulation of the desired products in the reaction.

[0272] The method relies on a DNA-based probe with a fluorescent reporter at one end and a quencher of fluorescence at the opposite end of the probe. The close proximity of the reporter to the quencher prevents detection of its fluorescence; breakdown of the probe by the 5' to 3' exonuclease activity of the Taq polymerase breaks the reporter-quencher proximity and thus allows unquenched emission of fluorescence, which can be detected after excitation with a laser. An increase in the product targeted by the reporter probe at each PCR cycle therefore causes a proportional increase in fluorescence due to the breakdown of the probe and release of the reporter.

[0273] The PCR is prepared as usual, and the reporter probe is added.

[0274] During the annealing stage of the PCR both probe and primers anneal to the DNA target.

[0275] Polymerisation of a new DNA strand is initiated from the primers, and once the polymerase reaches the probe, its 5'-3'-exonuclease degrades the probe, physically separating the fluorescent reporter from the quencher, resulting in an increase in fluorescence.

[0276] Fluorescence is detected and measured in the real-time PCR thermocycler, and its geometric increase corresponding to exponential increase of the product is used to determine the threshold cycle (CT) in each reaction.

[0277] In one particular embodiment, the measure of the expression level of the genes as defined above is achieved, in addition to the above defined specific oligonucleotides, by using a probe commercially available. Each gene for which the expression level is expected is associated with a specific probe, a probe recognizing one gene is not able to recognize another gene. Moreover a probe specific of one gene can also detect, when they exist, variants of said genes.

[0278] The advantageous probes used in the invention are listed in the table A.

[0279] The association between gene/variant/oligonucleotides and probes are represented in the following table 4.

TABLE-US-00004 TABLE 4 SEQ ID SEQ ID SEQ ID Gene name SEQ ID variant Oligo forward Oligo reverse SEQ probes GPX3 SEQ ID NO: 1 -- SEQ ID NO: 25 SEQ ID NO: 26 CCAGCCGC GPX1 (1) SEQ ID NO: 2 -- SEQ ID NO: 27 SEQ ID NO: 28 GGTGGTGG GLRX2 (2) SEQ ID NO: 3 -- SEQ ID NO: 29 SEQ ID NO: 30 GGCGGCGG CAT SEQ ID NO: 4 -- SEQ ID NO: 31 SEQ ID NO: 32 TGCTGGAG PRDX SEQ ID NO: 5 SEQ ID NO: 73 or SEQ ID NO: 33 SEQ ID NO: 34 CTGGCTGG (1-2-3) SEQ ID NO: 74 PRDX5 (2) SEQ ID NO: 6 -- SEQ ID NO: 35 SEQ ID NO: 36 GGAAGGAG SOD2 SEQ ID NO: 7 SEQ ID NO: 75 or SEQ ID NO: 37 SEQ ID NO: 38 CTGCTGGG (1-2-3) SEQ ID NO: 76 GSR SEQ ID NO: 8 SEQ ID NO: 77 or SEQ ID NO: 39 SEQ ID NO: 40 GCTGGAAG (1-2-3-4) SEQ ID NO: 78 or SEQ ID NO: 79 GLRX SEQ ID NO: 9 SEQ ID NO: 80 SEQ ID NO: 41 SEQ ID NO: 42 GGTGGCTG PRDX2 (1) SEQ ID NO: 10 -- SEQ ID NO: 43 SEQ ID NO: 44 TGGGGAAG PRDX5 SEQ ID NO: 11 SEQ ID NO: 81 or SEQ ID NO: 45 SEQ ID NO: 46 GGAAGGAG (1-3) SEQ ID NO: 82 SOD1 SEQ ID NO: 12 -- SEQ ID NO: 47 SEQ ID NO: 48 TGGGGAAG TXN SEQ ID NO: 13 -- SEQ ID NO: 49 SEQ ID NO: 50 CAGCAGCC PRDX3 SEQ ID NO: 14 SEQ ID NO: 83 SEQ ID NO: 51 SEQ ID NO: 52 CTGCTTCC (1-2) GPX7 SEQ ID NO: 15 -- SEQ ID NO: 53 SEQ ID NO: 54 GGAAGGAG GPX4 SEQ ID NO: 16 SEQ ID NO: 84 or SEQ ID NO: 55 SEQ ID NO: 56 CTGCCCCA (1-2-3) SEQ ID NO: 85 TXN2 SEQ ID NO: 17 -- SEQ ID NO: 57 SEQ ID NO: 58 GGCCCCAG PRDX4 SEQ ID NO: 18 -- SEQ ID NO: 59 SEQ ID NO: 60 ACTGGGAA GPX1(2) SEQ ID NO: 19 -- SEQ ID NO: 61 SEQ ID NO: 62 CTCCTCCT GLRX3 SEQ ID NO: 20 -- SEQ ID NO: 63 SEQ ID NO: 64 TGGTGGAA PRDX2 (3) SEQ ID NO: 21 -- SEQ ID NO: 65 SEQ ID NO: 66 GGAGGCTG PRDX6 SEQ ID NO: 22 -- SEQ ID NO: 67 SEQ ID NO: 68 CCTGGAGC GLRX5 SEQ ID NO: 23 -- SEQ ID NO: 69 SEQ ID NO: 70 TGCTGGAG GLRX2 (1) SEQ ID NO: 24 -- SEQ ID NO: 71 SEQ ID NO: 72 GGATGGAG Table 4

[0280] Table 4 can be read as follows: The expression level of GPX3 (SEQ ID NO: 1) can be quantitatively measured by using the oligonucleotides SEQ ID NO: 24 and 25, and using as probe, coupled with a fluorescent dye and a quencher, having the following sequence CCAGCCGC.

[0281] Also, another example: The expression level of PDRX (SEQ ID NO: 5), or one of its variants (SEQ ID NO 73 or 74) can be quantitatively measured by using the oligonucleotides SEQ ID NO: 33 and 34, and using as probe, coupled with a fluorescent dye and a quencher, having the following sequence CTGGCTGG.

[0282] The above definitions apply mutatis mutandis for the other genes.

[0283] Quencher and dye mentioned above can be chosen by the skilled person, depending of the assay.

[0284] In another advantageous embodiment, the invention relates to the method as defined above, using at least one of the oligonucleotide allowing the measurement of the expression level of each of at least 6 genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 1-6, preferably using at least one of the oligonucleotide allowing the measurement of the expression level of each of at least 10 genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 1-10, in particular using at least one of the oligonucleotide allowing the measurement of the expression level of each of the 24 genes comprising or being constituted by the nucleic acid sequences SEQ ID NO: 1-24.

[0285] In this embodiment, the method is adapted for northern blot assay.

[0286] The invention also relates to a composition comprising oligonucleotides allowing the measure of the expression of at least the genes of a sub-group of 6 genes belonging to a set of genes chosen among a group of 24 genes, said group of 24 genes comprising or being constituted by the nucleic acid sequences SEQ ID NO:1 to 24, said 6 genes belonging to said sub-group comprising or being constituted by the nucleic acid sequences SEQ ID NO: 1 to 6.

[0287] In one advantageous embodiment, the invention relates to a composition as defined above, comprising oligonucleotides allowing the measure of the expression of at least the genes of a set of 10 genes chosen among a group of 24 genes, said group of 24 genes comprising or being constituted by the nucleic acid sequences SEQ ID NO:1 to 24,

[0288] said 10 genes belonging to said set comprising or being constituted by the nucleic acid sequences SEQ ID NO: 1 to 10.

[0289] In one other advantageous embodiment, the invention relates to a composition as defined above, comprising oligonucleotides allowing the measure of the expression of the 24 of said group of 24 genes.

[0290] In one other advantageous embodiment, the invention relates to a composition as defined above, said composition comprising at least 12 oligonucleotides chosen among a library of 48 oligonucleotides comprising or consisting of the nucleic acid sequences SEQ ID NO: 25-72, allowing the measure of the expression of at least the genes of a sub-group of 6 genes belonging to a set of genes chosen among a group of 24 genes comprising or being constituted by the nucleic acid sequences SEQ ID NO:1 to 24,

said at least 12 oligonucleotides comprising or consisting of the nucleic acid sequences SEQ ID NO: 25-36.

[0291] In one other advantageous embodiment, the invention relates to a composition as defined above, said composition comprising at least 20 oligonucleotides chosen among a library of 48 oligonucleotides comprising or consisting of the nucleic acid sequences SEQ ID NO: 25-72, allowing the measure of the expression of at least the genes a set of 10 genes chosen among a group of 24 genes comprising or being constituted by the nucleic acid sequences SEQ ID NO:1 to 24,

said at least 20 oligonucleotides comprising or consisting of the nucleic acid sequences SEQ ID NO: 25-44.

[0292] In one other advantageous embodiment, the invention relates to a composition as defined above, said composition comprising 48 oligonucleotides comprising or consisting of the nucleic acid sequences SEQ ID NO: 25-72, allowing the measure of the expression of a group of 24 genes comprising or being constituted by the nucleic acid sequences SEQ ID NO:1 to 24.

[0293] The above composition may further comprise probes as defined above in Table 4.

[0294] In another aspect, the invention relates to the composition as mentioned above, for its use for the diagnosis, and/or the classification, preferably in vitro, of an hematological disorder, in particular myeloid and/or lymphoid hematological disorder, preferably myeloid hematological disorder.

[0295] Therefore, the invention relates to the above composition per se, and relates to said composition for its use as mentioned above.

[0296] The invention relates to a kit comprising at least 12 oligonucleotides chosen among a group of 48 oligonucleotides comprising or consisting of the nucleic acid sequences SEQ ID NO: 25-72, said at least 12 oligonucleotides comprising or consisting of the nucleic acid sequences SEQ ID NO: 25-36.

[0297] In one advantageous embodiment, the invention relates to the kit as defined above, comprising at least 20 oligonucleotides, said at least 20 oligonucleotides comprising or consisting of the nucleic acid sequences SEQ ID NO: 25-44.

[0298] In one advantageous embodiment, the invention relates to a kit as defined above comprising 48 oligonucleotides comprising or consisting of the nucleic acid sequences SEQ ID NO: 25-72.

[0299] The invention also relates to, in one advantageous embodiment, a kit as defined above, further comprising at least 6 specific probes that respectively interact the nucleic acid molecules comprising or consisting of SEQ ID NO: 1-6, preferably further comprising at least 10 specific probes that respectively interact the nucleic acid molecules comprising or consisting of SEQ ID NO: 1-10, in particular further comprising at least 24 specific probes that respectively interact the nucleic acid molecules comprising or consisting of SEQ ID NO: 1-24.

[0300] In one another advantageous embodiment, the invention relates to a kit as defined above, further comprising nucleic acid molecules corresponding to the genes SEQ ID NO: 1-24, in an amount representative at least one pathology chosen among: acute myeloid leukemia (AML), refractory anemia with ringed sideroblasts (RARS), refractory cytopenia with multilineage dysplasia (RCMD), refractory anemia with excess of blasts (RAEB) or 5q-syndrome and unclassifiable myelodysplasia.

[0301] In another advantageous embodiment, the invention also relates to the kit as defined above, further comprising nucleic acid molecules of a control sample as defined above.

[0302] The invention also relates to a positive control sample comprising or being constituted by at least the nucleic acid molecules corresponding to the genes represented by SEQ ID NO: 1-6, chosen among the group of 24 genes represented by SEQ ID NO: 1-24, said nucleic acid molecules being present in said sample in an amount as represented in the 6 first lines of table 5 or table 6, compared to an healthy sample in which each of the respective nucleic acid molecules are present in an amount of 1.

TABLE-US-00005 TABLE 5 Table 5 RARS RCMD RAEB AML SEQ ID NO: 1 1.32-2.62 0.01-1.67 0.43-2.88 0.03-0.30 SEQ ID NO: 2 2.32-2.98 0.12-9.95 4.48-18.89 2.98-6.98 SEQ ID NO: 3 3.74-9.79 0.08-10.50 0.14-10.87 3.12-15.80 SEQ ID NO: 4 1.28-2.20 0.02-3.94 0.99-8.75 0.32-10.20 SEQ ID NO: 5 1.25-3.68 0.18-5.77 4.59-10.04 2.00-13.34 SEQ ID NO: 6 1.84-6.09 0.00-10.24 0.05-5.53 1.72-5.56 SEQ ID NO: 7 0.65-0.95 0.54-4.37 0.85-2.13 0.08-0.79 SEQ ID NO: 8 0.67-1.09 0.00-1.18 0.44-1.15 0.20-0.58 SEQ ID NO: 9 0.90-1.25 0.43-2.05 0.07-1.05 0.08-0.79 SEQ ID NO: 0.48-1.87 0.02-2.13 0.49-4.41 0.04-0.25 10 SEQ ID NO: 0.81-1.69 0.00-2.01 1.19-1.47 0.65-1.76 11 SEQ ID NO: 0.64-2.12 0.23-3.74 1.48-2.74 0.45-2.94 12 SEQ ID NO: 0.96-2.43 0.63-4.99 1.14-4.22 0.42-5.34 13 SEQ ID NO: 1.20-2.02 0.16-3.16 0.63-1.47 0.70-2.15 14 SEQ ID NO: 0.63-1.44 0.09-1.18 0.00-1.37 0.55-3.71 15 SEQ ID NO: 0.94-2.53 0.10-3.29 2.58-6.22 1.35-4.25 16 SEQ ID NO: 1.07-2.22 0.15-2.58 2.30-2.85 0.90-3.89 17 SEQ ID NO: 1.48-2.79 0.37-2.86 3.31-4.99 1.10-6.67 18 SEQ ID NO: 1.04-5.32 0.49-3.87 1.96-8.77 0.64-5.18 19 SEQ ID NO: 1.49-5.23 0.09-2.58 1.81-11.93 0.84-7.09 20 SEQ ID NO: 0.93-8.83 0.01-2.87 0.15-3.76 0.08-0.63 21 SEQ ID NO: 0.81-3.63 0.14-36.64 0.25-12.98 0.98-5.93 22 SEQ ID NO: 1.47-3.45 0.19-7.04 2.80-53.30 0.26-3.65 23 SEQ ID NO: 0.78-2.25 0.35-1.60 0.40-0.87 0.18-0.83 24

[0303] Table 5 represents, for each gene of SEQ ID NO: 1-24, the specific interval corresponding to the mentioned pathology.

TABLE-US-00006 TABLE 6 Table 6 RARS RCMD RAEB AML SEQ ID 2.14 .+-. 0.71 0.76 .+-. 0.56 1.60 .+-. 1.23 0.16 .+-. 0.13 NO: 1 SEQ ID 2.64 .+-. 0.33 3.65 .+-. 3.95 18.89 .+-. 17.93 4.95 .+-. 1.70 NO: 2 SEQ ID 5.97 .+-. 3.33 4.04 .+-. 3.49 5.47 .+-. 5.36 9.73 .+-. 5.24 NO: 3 SEQ ID 1.63 .+-. 0.50 1.46 .+-. 1.64 4.31 .+-. 4.00 4.01 .+-. 3.73 NO: 4 SEQ ID 2.52 .+-. 1.22 1.89 .+-. 2.14 7.40 .+-. 2.73 5.03 .+-. 4.74 NO: 5 SEQ ID 4.55 .+-. 2.36 3.64 .+-. 4.01 5.30 .+-. 5.13 4.19 .+-. 1.67 NO: 6 SEQ ID 0.84 .+-. 0.17 1.80 .+-. 1.61 1.32 .+-. 0.70 0.33 .+-. 0.27 NO: 7 SEQ ID 0.85 .+-. 0.22 0.76 .+-. 0.42 0.78 .+-. 0.35 0.40 .+-. 0.18 NO: 8 SEQ ID 1.07 .+-. 0.18 1.22 .+-. 0.67 0.54 .+-. 0.49 0.45 .+-. 0.26 NO: 9 SEQ ID 1.06 .+-. 0.72 0.74 .+-. 0.73 2.40 .+-. 1.96 0.11 .+-. 0.08 NO: 10 SEQ ID 1.33 .+-. 0.46 0.93 .+-. 0.83 1.47 .+-. 0.29 1.11 .+-. 0.44 NO: 11 SEQ ID 1.31 .+-. 0.75 1.12 .+-. 1.30 1.92 .+-. 0.71 1.27 .+-. 0.97 NO: 12 SEQ ID 1.51 .+-. 0.80 2.06 .+-. 1.70 2.32 .+-. 1.67 2.52 .+-. 1.87 NO: 13 SEQ ID 1.68 .+-. 0.43 1.64 .+-. 1.21 1.17 .+-. 0.46 1.70 .+-. 0.59 NO: 14 SEQ ID 0.99 .+-. 0.41 0.79 .+-. 0.39 0.70 .+-. 0.68 2.57 .+-. 1.32 NO: 15 SEQ ID 1.61 .+-. 0.82 1.58 .+-. 1.28 6.22 .+-. 4.17 2.62 .+-. 1.37 NO: 16 SEQ ID 1.69 .+-. 0.58 1.15 .+-. 0.96 2.79 .+-. 0.47 2.40 .+-. 1.33 NO: 17 SEQ ID 2.17 .+-. 0.65 1.19 .+-. 0.88 4.36 .+-. 0.91 2.59 .+-. 2.33 NO: 18 SEQ ID 2.59 .+-. 2.37 1.67 .+-. 1.25 4.89 .+-. 3.50 2.12 .+-. 1.80 NO: 19 SEQ ID 2.75 .+-. 2.15 1.27 .+-. 1.12 5.31 .+-. 5.74 3.96 .+-. 2.61 NO: 20 SEQ ID 3.80 .+-. 4.37 1.38 .+-. 1.18 2.41 .+-. 1.97 0.36 .+-. 0.23 NO: 21 SEQ ID 1.83 .+-. 1.56 7.33 .+-. 14.42 5.69 .+-. 6.56 2.90 .+-. 2.21 NO: 22 SEQ ID 2.76 .+-. 1.12 2.60 .+-. 2.88 21.67 .+-. 27.56 1.49 .+-. 1.34 NO: 23 SEQ ID 1.56 .+-. 0.74 0.95 .+-. 0.46 0.87 .+-. 0.62 0.41 .+-. 0.25 NO: 24

[0304] Table 6 represents, for each gene of SEQ ID NO: 1-24, the mean.+-.the standard deviation corresponding to the mentioned pathology.

[0305] The invention also relates to a method for determining the efficacy of a treatment of an hematological disorder, said treatment being liable to be administered to a patient,

said method comprising a). a step of contacting, preferably in vitro, a biological sample of a subject afflicted by an hematological disorder, preferably a myeloid and/or a lymphoid hematological disorder, more preferably myeloid hematological disorder, with a drug liable to be used to treat, or liable to treat, said hematological disorder, b). a step of measuring, in the biological sample contacted with a drug in step a)., the expression level of at least the genes of a sub-group of 6 genes belonging to a set of genes chosen among a group of 24 genes, said group of 24 genes comprising or being constituted by the nucleic acid sequences SEQ ID NO:1 to 24, said 6 genes belonging to said sub-group comprising or being constituted by the nucleic acid sequences SEQ ID NO: 1 to 6, and c). a step of comparing the expression level of said at least the genes of a sub-group of 6 genes belonging to a set of genes chosen among a group of 24 genes obtained in step b). with the expression level of said at least the genes of a sub-group of 6 genes belonging to a set of genes chosen among a group of 24 genes obtained measured in said biological sample which has not been contacted with said drug liable to be used to treat, or liable to treat, said hematological disorder.

[0306] The above method is easy to carry out, and allows to evaluate the AML sample susceptibility to a drug. This is very important to reduce the cost of treatments, that can be ineffective in patient, because the tumor is resistant to the drug.

[0307] The above method is advantageously used to screen, in vitro, drugs having an effect on AML progression, and that could be used in vivo for the treatment of the patient.

[0308] This is more advantageous important for screening drugs, or compounds that are able to modulate the epigenetic modification, in particular demethylating agents such as azacytidine (5-azacytidine) or decitabine (5-azadeoxycytidine).

[0309] Azacytidine and decitabine are powerful chemotherapeutic agents used for treating AML and high grade MDS, but only about 30% of AML and high grade MDS are sensitive to their effects. Thus, in order to reduce the costs, and the side effects of ineffective treatment, it is advantageous to verify in vitro, if the AML and high grade MDS that has to be treated is responsive to these compounds.

[0310] The example 9 shows that the treatment of AML sample with azacytidine can modulate the expression ratio of the genes SEQ ID NO: 1-24, (and consequently at the genes SEQ ID NO: 1-6) demonstrating that azacytidine, in this particular patient from which the AML sample derives, would be effective if it is used in vivo.

[0311] Advantageously, the invention relates to a method as defined above, wherein said set consists of 10 genes consisting of the nucleic acid sequences SEQ ID NO: 1-10.

[0312] Advantageously, the invention relates to a method as defined above, the expression level of the 24 genes consisting of SEQ ID NO: 1-24 is measured.

LEGEND OF THE FIGURES

[0313] FIG. 1 represents the natural mechanism used in cell for eliminating reactive oxide species (ROS, O.sub.2.sup..cndot.-). O.sub.2.sup..cndot.- were converted into H.sub.2O.sub.2 molecules, said H.sub.2O.sub.2 being themselves either converted into H.sub.2O and O.sub.2 as a detoxifying process, or into .sup..cndot.OH and OH.sup.- that exert biological effects in cells.

[0314] FIG. 2 represents the imbrications between the subgroup (A), the set (B) and the group (C) of the genes according to the invention. D represents the ensemble corresponding to the genes belonging to the set but that do not belong to the sub-group. E represents the ensemble corresponding to the genes belonging to the group but that do not belong to the set.

[0315] FIGS. 3 A-E represent the schematic representation of the expression level of each genes represented by SEQ ID NO: 1-24, for each pathologies: RARS, RCMD, RAEB and AML.

[0316] FIG. 3A represents an histogram showing the expression level of the genes indicated in the x-axis, obtained by qRT-PCR, compared to the housekeeping gene GAPDH, by using 5 healthy bone marrow samples (control sample).

[0317] Y-axis represents the ratio .DELTA.CT.sub.gene (.DELTA.CT.sub.gene=CT.sub.gene-CT.sub.GAPDH).

[0318] FIG. 3B is a graphic representation of the mean (black line) and standard error of the mean (grey area) of the variation of expression of each indicated genes (x-axis) in a RARS samples compared to control samples (healthy bone marrow samples). Y-axis represents the variation of the amount compared to control samples, expressed in log.sub.10.

[0319] FIG. 3C is a graphic representation of the mean (black line) and standard error of the mean (grey area) of the variation of expression of each indicated genes (x-axis) in RCMD samples compared to control samples (healthy bone marrow samples). Y-axis represents the variation of the amount compared to control samples, expressed in log.sub.10.

[0320] FIG. 3D is a graphic representation of the mean (black line) and standard error of the mean (grey area) of the variation of expression of each indicated genes (x-axis) in RAEB samples compared to control samples (healthy bone marrow samples). Y-axis represents the variation of the amount compared to control samples, expressed in log.sub.10.

[0321] FIG. 3E is a graphic representation of the mean (black line) and standard error of the mean (grey area) of the variation of expression of each indicated genes (x-axis) in AML samples compared to control samples (healthy bone marrow samples). Y-axis represents the variation of the amount compared to control samples, expressed in log.sub.10.

[0322] FIGS. 4A-D represent the schematic representation of the expression level of each of the genes represented by SEQ ID NO: 1-24, for each pathologies: RARS (FIG. 4A), RCMD (FIG. 4B), RAEB (FIG. 4C) and AML (FIG. 4D). Representation of the expression level of each genes represented by SEQ ID NO: 1-24 of an unclassifiable myelodysplastic syndroma sample is represented in each FIGS. 4A-D by hashed line.

[0323] FIGS. 5A and B represent the schematic representation of the expression level of each of the genes represented by SEQ ID NO: 1-24 of two independent unclassifiable myelodysplastic syndroma samples (FIG. 5B), said expression being similar to the expression level observed in RCMD samples (FIG. 5A)

[0324] FIG. 6 represents the schematic representation of the expression level of each of the genes represented by SEQ ID NO: 1-24 of two independent chronic myelomonocytic leukemia samples (FIG. 6B) said expression being different from the expression level observed in RAEB samples (FIG. 6A)

[0325] FIG. 7 represents the schematic representation of the expression level of each of the genes represented by SEQ ID NO: 1-24 of a patient at diagnosis of RCMD (panel B) and after 12 months (panel B, hatched line). The sample at diagnosis is similar to RCMD sample (panel A) and the sample after 12 months has acquired characteristics of RAEB (see PDRX2(1) and GLRX5 in panel B and panel C).

[0326] FIG. 8 represents the schematic representation of the expression level of each of the genes represented by SEQ ID NO: 1-24 of AML, compared to same AML after treatment with azacytidine (hatched line).

EXAMPLES

Preliminary Comment

[0327] All the samples used in the following examples have been first tested according to the invention, in a blind test, and compared with control samples.

[0328] All the SMD and leukemic samples satisfy the provisions of the method according to the invention, i.e. in each of the samples at least 3 genes of the genes SEQ ID NO: 1-6 are expressed such that their ratio compared to the expression of the same corresponding genes in control samples is either lower than 0.5 or higher to 2.

Example 1

[0329] In the following examples, the patient's samples and the analysis genes expressions are analysed as follows.

Material and Methods

Sample Harvest

[0330] Normal bone marrow (BM) samples (used as a reference) were obtained from patients undergoing orthopedic surgery. Bone marrow samples from MDS and AML patients were obtained at the time of diagnosis and during the follow up. AML and MDS cells were classified according to morphological, cytochemical and cytogenetical findings. Patients were informed and consenting following a procedure approved by the ethical committee. BM samples were aspirated into heparinized syringes and transferred to EDTA tubes.

Erythrocyte Lysis

[0331] Lysis was performed in 47 mL lysis buffer containing EDTA (0.12 mM), potassium bicarbonate (KHCO.sub.3) (10 mM), and ammonium chloride (NH.sub.4Cl) (150 mM) for 3 mL of BM. Following incubation at room temperature for 15 min, cells were centrifuged at 700 g for 10 min, and washed, twice, in 20 mL of Phosphate Buffered Saline (PBS) (Invitrogen). The pellet was resuspended in Trizol.RTM. (Invitrogen) (1 mL/8.10.sup.6 cells), mixed vigorously for 15 minutes. The lysate was stored at -80.degree. C. until the RNA extraction step.

RNA Extraction

[0332] Total RNA extraction was performed according to the chloroform/isopropanol/ethanol method. The phase separation was obtained by adding chloroform (0.2 mL per 1 mL of TRIzol.RTM. purchased from Invitrogen) to the lysate. After mixing for 50 sec, the solution was separated into three phases by centrifugation at 12,000 g for 15 min at 4.degree. C. RNA was precipitated, from the aqueous phase, by adding isopropanol (0.5 mL per 1 mL of TRIzol.RTM.). Following incubation at room temperature for 10 min and centrifugation at 12,000 g for 10 min at 4.degree. C., RNA was washed in 75% ethanol (1 mL per 1 mL of TRIzol.RTM.) and centrifugated at 7500 g for 5 min at 4.degree. C. This step was performed twice. After removing ethanol supernatant, the RNA pellet was air-dried for 20 min. Then, RNA was dissolved in 50 .mu.L of UltraPure.TM. DEPC-treated water (Invitrogen) and stored at -80.degree. C.

RNA Quantification and Qualification

[0333] Total cellular RNA was quantified using a Nano-Drop 1000 spectrophotometer (Nano-Drop Technologies) and RNA purity was analyzed using an Agilent 2100 Bioanalyzer (Agilent Technologies).

Reverse Transcription and Quantitative Real-Time PCR (qRT-PCR) Analysis

[0334] Three micrograms of total RNA from each sample were reverse transcribed using the SuperScript.RTM. VILO.TM. cDNA Synthesis kit (Invitrogen) according to the protocol of the supplier. The relative quantification of gene expression was done by real-time PCR on the LightCycler.RTM. 480 microwell plate-based cycler platform (Roche Applied Science) using Universal ProbeLibrary assays designed with the ProbeFinder software (Roche Applied Science, www.roche-applied-science.com/sis/rtpcr/upl/ezhome.html). Primers were purchased from Invitrogen and Universal ProbeLibrary probes from Roche Applied Science. The nucleotide sequences of the primers and probes of each target are shown in Table A. All targets were concomitantly analyzed. qRT-PCR reactions were carried out in a total volume of 10 .mu.L on 20 ng of cDNA using LightCycler.RTM. 480 Probes Master (Roche Applied Science). The LightCycler.RTM. 480 was programmed to an initial denaturation (95.degree. C., 10 min) following by 45 cycles of 10 sec at 95.degree. C., 30 sec at 60.degree. C., 1 sec at 72.degree. C. and a final cooling step at 40.degree. C. for 10 sec. All reactions were run in triplicate, and average values were used for quantification. Results were analyzed by the relative quantification method (.DELTA..DELTA.CT=.DELTA.CT.sub.patient-.DELTA.CT.sub.reference) using the Cycle threshold (CT) values determined with the LightCycler.RTM. 480 software (release 1.5.0) from Roche Applied Science. The human glyceraldehyde-3-phosphate dehydrogenase gene (GAPDH) was used as the endogenous control to normalize the expression of the target (.DELTA.CT=CT.sub.target-CT.sub.endogenous control). Change of relative mRNA expression between a patient and the reference was determined for each target using the 2exp(-.DELTA..DELTA.CT) method (Livak K J and Schmittgen T D. Analysis of relative gene expression data using real time quantitative PCR and the 2exp(-.DELTA..DELTA.CT) method. Methods 2001; 25:402-408).

Example 2

Use of 6 Genes for Diagnosing Myelodysplasia and Leukemia

[0335] In order to test the validity of the method, patient samples identified as representative of leukemic samples where used, and the expression level of each of the genes represented by SEQ ID NO: 1 to SEQ ID NO: 6 was evaluated by RT-qPCR as disclosed in Example 1, by using oligonucleotides and probes of the Table 4.

[0336] The results are represented in the following table 7:

TABLE-US-00007 TABLE 7 AML AML # 1 AML # 2 # 3 AML # 4 AML # 5 SEQ ID NO: 1 0.03 0.29 0.30 0.08 0.08 SEQ ID NO: 2 3.54 6.16 6.98 2.98 5.10 SEQ ID NO: 3 14.24 7.07 8.41 15.80 3.12 SEQ ID NO: 4 10.20 4.06 3.32 2.17 0.32 SEQ ID NO: 5 13.34 2.96 2.00 4.46 2.36 SEQ ID NO: 6 5.56 3.22 5.12 5.34 1.72

[0337] Table 7 represents the ratio of the expression level of each indicated genes (SEQ ID NO: 1-6) for each AML sample #1-#5

[0338] The criterions defined in the invention are satisfied:

A sample is considered as representative of an hematological disorder when the ratio of the expression level of each genes of any combination of 3 genes among the genes represented by SEQ ID NO: 1-6 is either .gtoreq.2.0 or .ltoreq.0.5.

[0339] Whatever the combination of 3 genes taken in consideration, the above criterions are satisfied.

[0340] The other criterion is that: if ratio of SEQ ID NO: 1 is .ltoreq.0.3 and the ratio of both SEQ ID NO: 2 and 3 are .gtoreq.3.0, the sample is representative of an AML.

[0341] Again, for all the 5 above AML samples tested (Table 7), the criterions are satisfied.

[0342] All the tested samples satisfy the criterions regarding AML. Therefore, the method according to the invention allows the discrimination between AML and myelodysplastic disorders.

Example 3

Validation of the Method with Leukemic Cell Lines

[0343] In order to validate the method, the expression level of the genes represented by SEQ ID NO: 1-6 was evaluated in 11 cell lines corresponding to almost all the AML subtypes defined according to the FAB classification.

[0344] The following cell lines have been tested:

KG1a and KG1 (FAB M0/M1), HL60 (FAB M2), KASUMI-1 (FAB M2), ML-2 (FAB M4), MV4-11 (FAB M5), THP-1 (FAB M5), U937 (FAB M5), K562 (FAB M6), TF-1 (FAB M6) and UT7 (FAB M7).

[0345] The expression levels are indicated in the following table 8.

TABLE-US-00008 TABLE 8 SEQ ID MV4- NO: KG1a KG1 HL60 Kasumi-1 ML-2 11 THP-1 U937 K562 TF-1 UT7 1 0.05 0.06 0.05 0.04 0.06 0.03 0.15 0.07 0.10 0.03 0.30 2 3.42 5.26 4.74 6.23 5.61 17.95 45.70 12.20 4.58 7.92 48.90 3 37.71 54.38 33.49 14.34 34.17 17.19 77.40 45.84 69.24 18.38 53.66 4 0.53 0.91 2.73 1.25 2.13 5.19 8.80 1.91 2.08 1.54 2.44 5 197.83 39.98 14.21 9.46 6.10 8.08 29.76 17.93 48.00 31.97 33.70 6 30.91 32.92 13.85 12.11 9.14 5.84 20.37 10.29 24.11 13.99 30.32

[0346] Table 8 represents the ratios R.sub.i between the expression levels of the indicated gene in the corresponding cell line, compared to the expression levels of the corresponding gene in control samples (healthy bone marrow samples).

[0347] The criterions defined in the invention as satisfied:

A sample is considered as representative of an hematological disorder when the ratio of the expression level of each genes of any combination of 3 genes among the genes represented by SEQ ID NO: 1-6 is either .gtoreq.2.0 or .ltoreq.0.5.

[0348] Whatever the combination of 3 genes taken in consideration, the above criterions are satisfied.

[0349] The other criterion is that: if ratio of SEQ ID NO: 1 is .ltoreq.0.3 and the ratio of both SEQ ID NO: 2 and 3 are .gtoreq.3.0, the sample is representative of an AML.

[0350] Again, for all the 11 above cell lines tested (Table 3), the criterions are satisfied, and the method confirm that these results obtained with this cell lines correspond to those obtained with primary leukemic cells.

Example 4

Classification of Myelodyplastic Sample

[0351] As defined above, it has been proposed that the measure of the expression level of the genes represented by SEQ ID NO: 1-24 allows to identify subtype of myelodysplastic syndrome.

[0352] A panel of patient sample, classified by other techniques, has been tested to validate the method according to the invention.

[0353] 3 RARS, 6 RCMD and 3 RAEB have been used, and the expression level of the genes represented by SEQ ID NO: 1-24 have been evaluated.

[0354] Results are presented hereafter:

TABLE-US-00009 TABLE 9 Table 9: RARS samples RARS RARS RARS #1 #2 #3 SEQ ID NO: 1 2.48 1.32 2.62 SEQ ID NO: 2 2.63 2.32 2.98 SEQ ID NO: 3 9.79 3.74 4.38 SEQ ID NO: 4 2.20 1.41 1.28 SEQ ID NO: 5 3.68 2.64 1.25 SEQ ID NO: 6 5.72 6.09 1.84 SEQ ID NO: 7 0.65 0.93 0.95 SEQ ID NO: 8 1.09 0.79 0.67 SEQ ID NO: 9 1.25 1.06 0.90 SEQ ID NO: 10 1.87 0.48 0.82 SEQ ID NO: 11 1.69 1.50 0.81 SEQ ID NO: 12 2.12 0.64 1.17 SEQ ID NO: 13 2.43 1.16 0.96 SEQ ID NO: 14 2.02 1.82 1.20 SEQ ID NO: 15 1.44 0.89 0.63 SEQ ID NO: 16 2.53 1.35 0.94 SEQ ID NO: 17 2.22 1.07 1.79 SEQ ID NO: 18 2.79 2.25 1.48 SEQ ID NO: 19 1.43 5.32 1.04 SEQ ID NO: 20 5.23 1.49 1.53 SEQ ID NO: 21 1.64 8.83 0.93 SEQ ID NO: 22 3.63 1.06 0.81 SEQ ID NO: 23 3.35 3.45 1.47 SEQ ID NO: 24 1.64 0.78 2.25

TABLE-US-00010 TABLE 10 RCMD samples RCMD RCMD RCMD RCMD RCMD RCMD #1 #2 #3 #4 #4 #5 SEQ ID 0.55 1.05 1.67 0.66 0.61 0.01 NO: 1 SEQ ID 0.52 9.95 5.75 0.54 0.12 5.05 NO: 2 SEQ ID 3.44 4.57 10.50 2.71 0.08 2.96 NO: 3 SEQ ID 2.07 2.53 3.94 0.06 0.02 0.13 NO: 4 SEQ ID 2.43 5.77 2.23 0.48 0.18 0.28 NO: 5 SEQ ID 3.96 6.09 10.24 1.46 0.10 0.00 NO: 6 SEQ ID 0.58 3.22 4.37 1.27 0.54 0.82 NO: 7 SEQ ID 0.91 1.05 1.18 0.84 0.57 0.00 NO: 8 SEQ ID 1.48 2.05 1.86 0.88 0.62 0.43 NO: 9 SEQ ID 0.68 2.13 0.75 0.31 0.53 0.02 NO: 10 SEQ ID 0.71 2.01 1.89 0.69 0.30 0.00 NO: 11 SEQ ID 0.72 3.74 0.78 0.55 0.71 0.23 NO: 12 SEQ ID 1.98 4.99 2.99 1.09 0.63 0.66 NO: 13 SEQ ID 1.81 3.16 2.89 0.69 1.10 0.16 NO: 14 SEQ ID 0.87 1.18 0.95 1.01 0.64 0.09 NO: 15 SEQ ID 1.43 3.29 2.80 0.35 0.10 1.48 NO: 16 SEQ ID 1.24 2.58 1.95 0.70 0.15 0.30 NO: 17 SEQ ID 1.08 2.86 1.23 0.37 0.95 0.64 NO: 18 SEQ ID 1.58 3.87 2.28 0.49 0.71 1.10 NO: 19 SEQ ID 2.58 2.46 1.71 0.47 0.09 0.31 NO: 20 SEQ ID 1.96 2.40 2.87 0.56 0.47 0.01 NO: 21 SEQ ID 0.59 3.60 2.49 0.51 0.14 36.64 NO: 22 SEQ ID 2.46 7.04 5.12 0.40 0.40 0.19 NO: 23 SEQ ID 0.81 1.60 1.37 0.67 0.93 0.35 NO: 24

TABLE-US-00011 TABLE 11 Table 11: RAEB samples RAEB #1 RAEB #2 RAEB #3 SEQ ID NO: 1 1.49 0.43 2.88 SEQ ID NO: 2 38.96 13.21 4.48 SEQ ID NO: 3 5.40 10.87 0.14 SEQ ID NO: 4 3.18 0.99 8.75 SEQ ID NO: 5 4.59 7.56 10.04 SEQ ID NO: 6 10.31 5.53 0.05 SEQ ID NO: 7 0.85 2.13 0.99 SEQ ID NO: 8 0.76 1.15 0.44 SEQ ID NO: 9 0.50 1.05 0.07 SEQ ID NO: 10 2.30 0.49 4.41 SEQ ID NO: 11 1.76 1.19 1.46 SEQ ID NO: 12 1.48 2.74 1.54 SEQ ID NO: 13 1.59 1.14 4.22 SEQ ID NO: 14 0.63 1.47 1.40 SEQ ID NO: 15 0.72 1.37 0.00 SEQ ID NO: 16 10.76 5.31 2.58 SEQ ID NO: 17 3.23 2.85 2.30 SEQ ID NO: 18 3.31 4.99 4.78 SEQ ID NO: 19 1.96 3.95 8.77 SEQ ID NO: 20 2.18 11.93 1.81 SEQ ID NO: 21 3.32 3.76 0.15 SEQ ID NO: 22 3.83 12.98 0.25 SEQ ID NO: 23 8.91 2.80 53.30 SEQ ID NO: 24 1.58 0.63 0.40

[0355] All the tested samples satisfy the criterions defined in the method according to the invention.

Example 5

Schematic Representation of RARS, RCMD, RAEB and AML Specific Profiles

[0356] The results corresponding to the above classification can be illustrated by an "antioxidogram", corresponding to areas representatives of a determined sample.

[0357] The expression of each gene of SEQ ID NO: 1-24 is measured by qRT-PCR as defined above, and compared to the expression level of an housekeeping gene GAPDH.

[0358] FIG. 3A represents the variation of expression of each genes of SEQ ID NO: 1-24 compared to GAPDH, classified according to their expression lever. The gene number is easily found by using the above Table 1.

[0359] The antioxidogram for RARS sample (FIG. 3B), RCMD sample (FIG. 3C), RAEB samples (FIG. 3D) and AML samples (3E) can be used to classify a new sample.

[0360] Indeed, by measuring the ratio for each genes represented by SEQ ID NO: 1-24, a curve can be drawn. This curve can then be compared to the antioxidograms, and thus, it is easy to determine into what type of pathology belongs the studied sample.

Example 6

Classification of Unclassifiable Myelodysplastic Syndromes

[0361] The above antioxidogram can be used to classify myelodysplastic syndromes for which other classification techniques fails.

[0362] The expression level of each of the genes of SEQ ID NO: 1-24 has been measured according to the invention, and an curve has been established.

[0363] This curve (hashed line) has been then compared to the "specific" antioxidogram of RARS, RCDM, RAEB and AML.

[0364] FIG. 4 shows that most of the ratios R.sub.i of expression are close to those that are characteristic of an RCDM sample.

[0365] FIG. 5 shows that two independent samples of unclassifiable myelodysplastic syndrome presents similar expression level of the genes of SEQ ID NO: 1-24, and thus are close to those that are characteristic of an RCDM sample.

Example 7

Distinction of MDS by Using the Oxydograms

[0366] Chronic myelomonocytic leukemia (CMML) is a form of leukemia featuring monocytosis. The categorization of this disease has been controversial.

[0367] Patients with CMML can present with various clinical features, mimicking either myelodysplastic syndroms or myeloproliferative neoplasms depending upon a patient's specific presentation.

[0368] Due to this controversy it was classified by the World Health Organization in a "myelodysplastic/myeloproliferative" category of medical conditions in the early 2000s.

[0369] The oxydogram according to the invention can be helpful to determine the status of a CMML sample, which is close to RAEB by histological analysis.

[0370] As shown in FIG. 6, the expression level of the genes SEQ ID NO: 1-24 of samples of two patients with CMML is different from the expression level of the genes SEQ ID NO: 1-24 of RAEB.

[0371] These data demonstrate that CMML are distinct from SDM.

Example 8

Use of Oxydogram for the Follow-Up of Evolutive MDS

[0372] As mentioned previously, MDS evolve progressively toward AML. It is thus important to know if this progression is slow or rapid.

[0373] The oxydogram can be used to determine if the molecular expression of the genes SEQ ID NO: 1-24 has evolved from diagnosis to a determined date.

[0374] An example is shown in FIG. 7. A patient has been diagnosed at t=0, as having a RCMD.

[0375] Twelve months from the diagnosis, the expression level of the genes SEQ ID NO: 1-24 have also been measured.

[0376] The FIG. 7 shows a difference between the expression at the diagnosis and after twelve months. After twelve months, the oxydogram shows that the patient is becoming to evolved from RCMD toward RAEB, although the histological analysis does not any differences.

Example 9

In Vitro Measure of Antitumoral Effect of Demethylating Agent

[0377] New therapeutic agents are actively searched in order to treat AML. These compounds are expensive, due to extensive searches, and unfortunately are not universally effective in all AML samples.

[0378] An average of 30% of AML subtypes are responsive to the new therapeutic drugs, and could effectively be used in the patient, in order to slow down the leukemic progression.

[0379] However, positive or negative in vivo response to a drug cannot be obtained before 6 to 9 months after the beginning of the treatment.

[0380] The oxydogram according to the invention can be used to evaluate, in vitro, if the drug will be effective on the leukemic sample, by studying the modulation of the expression level of the genes SEQ ID NO: 1-24.

[0381] FIG. 8 show an example of an AML sample treated with the demethylating agent azacytidine. This figure demonstrates that, in this specific sample, treatment modify the expression level of the genes, and could be efficient in vivo for the patient.

Sequence CWU 1

1

8511779DNAHomo sapiens 1gtcgccccgg gacggggagg tggggagctg agggcaagtc gcgcccgccc ctgaaatccc 60agccgcctag cgattggctg caagggtctc ggcttggccg cggattggtc acacccgagg 120gcttgaaagg tggctgggag cgccggacac ctcagacgga cggtggccag ggatcaggca 180gcggctcagg cgaccctgag tgtgccccca ccccgccatg gcccggctgc tgcaggcgtc 240ctgcctgctt tccctgctcc tggccggctt cgtctcgcag agccggggac aagagaagtc 300gaagatggac tgccatggtg gcataagtgg caccatttac gagtacggag ccctcaccat 360tgatggggag gagtacatcc ccttcaagca gtatgctggc aaatacgtcc tctttgtcaa 420cgtggccagc tactgaggcc tgacgggcca gtacattgaa ctgaatgcac tacaggaaga 480gcttgcacca ttcggtctgg tcattctggg ctttccctgc aaccaatttg gaaaacagga 540accaggagag aactcagaga tccttcctac cctcaagtat gtccgaccag gtggaggctt 600tgtccctaat ttccagctct ttgagaaagg ggatgtcaat ggagagaaag agcagaaatt 660ctacactttc ctaaagaact cctgtcctcc cacctcggag ctcctgggta catctgaccg 720cctcttctgg gaacccatga aggttcacga catccgctgg aactttgaga agttcctggt 780ggggccagat ggtataccca tcatgcgctg gcaccaccgg accacggtca gcaacgtcaa 840gatggacatc ctgtcctaca tgaggcggca ggcagccctg ggggtcaaga ggaagtaact 900gaaggccgtc tcatcccatg tccaccatgt aggggaggga ctttgttcag gaagaaatcc 960gtgtctccaa ccacactatc tacccatcac agaccccttt cctatcactc aaggccccag 1020cctggcacaa atggatgcat acagttctgt gtactgccag gcatgtgggt gtgggtgcat 1080gtgggtgttt acacacatgc ctacaggtat gcgtgattgt gtgtgtgtgc atgggtgtac 1140agccacgtgt ctacctatgt gtctttctgg gaatgtgtac catctgtgtg cctgcagctg 1200tgtagtgctg gacagtgaca accctttctc tccagttctc cactccaatg ataatagttc 1260acttacacct aaacccaaag gaaaaaccag ctctaggtcc aattgttctg ctctaactga 1320tacctcaacc ttggggccag catctcccac tgcctccaaa tattagtaac tatgactgac 1380gtccccagaa gtttctgggt ctaccacact ccccaacccc ccactcctac ttcctgaagg 1440gccctcccaa ggctacatcc ccaccccaca gttctccctg agagagatca acctccctga 1500gatcaaccaa ggcagatgtg acagcaaggg ccacggaccc catggcaggg gtggcgtctt 1560catgagggag gggcccaaag cccttgtggg cggacctccc ctgagcctgt ctgaggggcc 1620agcccttagt gcattcaggc taaggcccct gggcagggat gccacccctg ctccttcgga 1680ggacgtgccc tcacccctca ctggtccact ggcttgagac tcaccccgtc tgcccagtaa 1740aagcctttct gcagcagctg aaaaaaaaaa aaaaaaaaa 17792921DNAHomo sapiens 2cagttaaaag gaggcgcctg ctggcctccc cttacagtgc ttgttcgggg cgctccgctg 60gcttcttgga caattgcgcc atgtgtgctg ctcggctagc ggcggcggcg gcggcggccc 120agtcggtgta tgccttctcg gcgcgcccgc tggccggcgg ggagcctgtg agcctgggct 180ccctgcgggg caaggtacta cttatcgaga atgtggcgtc cctctgaggc accacggtcc 240gggactacac ccagatgaac gagctgcagc ggcgcctcgg accccggggc ctggtggtgc 300tcggcttccc gtgcaaccag tttgggcatc aggagaacgc caagaacgaa gagattctga 360attccctcaa gtacgtccgg cctggtggtg ggttcgagcc caacttcatg ctcttcgaga 420agtgcgaggt gaacggtgcg ggggcgcacc ctctcttcgc cttcctgcgg gaggccctgc 480cagctcccag cgacgacgcc accgcgctta tgaccgaccc caagctcatc acctggtctc 540cggtgtgtcg caacgatgtt gcctggaact ttgagaagtt cctggtgggc cctgacggtg 600tgcccctacg caggtacagc cgccgcttcc agaccattga catcgagcct gacatcgaag 660ccctgctgtc tcaagggccc agctgtgcct agggcgcccc tcctaccccg gctgcttggc 720agttgcagtg ctgctgtctc gggggggttt tcatctatga gggtgtttcc tctaaaccta 780cgagggagga acacctgatc ttacagaaaa taccacctcg agatgggtgc tggtcctgtt 840gatcccagtc tctgccagac caaggcgagt ttccccacta ataaagtgcc gggtgtcagc 900agaaaaaaaa aaaaaaaaaa a 9213735DNAHomo sapiens 3gcggtgtccg gcagtagagc tcgctgcaga tccgggctct gaccatgatt tggcgccgcg 60cggcgctggc ggggacgcgg ctggtttgga gcaggagcgg ctcggcaggc tggcttgaca 120gggcggcggg agctgcggga gctgcggcag ctgcggcctc tgggatggag agcaatacat 180catcatcttt ggagaattta gcgacggcgc ctgtgaacca gatccaagaa acaatttctg 240ataattgtgt ggtgattttc tcaaaaacat cctgttctta ctgtacaatg gcaaaaaagc 300ttttccatga catgaatgtt aactataaag tggtggaact ggacctgctt gaatatggaa 360accagttcca agatgctctt tacaaaatga ctggtgaaag aactgttcca agaatatttg 420tcaatggtac ttttattgga ggtgcaactg acactcatag gcttcacaaa gaaggaaaat 480tgctcccact agttcatcag tgttatttaa aaaaaagtaa gaggaaagaa tttcagtgat 540gtttatacta ataagtttgc tagtacagtg tcagttattt aaagtggtaa tgcccgataa 600tgtcttttaa atgtttgagg atgttttaaa tacatgcatt gtcttcacga agaagatgta 660aaaataatga acaataaatt gcggtggaaa cctcaaaaaa aaaaaaaaaa aaaaaaaaaa 720aaaaaaaaaa aaaaa 73542300DNAHomo sapiens 4actcggggca acaggcagat ttgcctgctg agggtggaga cccacgagcc gaggcctcct 60gcagtgttct gcacagcaaa ccgcacgcta tggctgacag ccgggatccc gccagcgacc 120agatgcagca ctggaaggag cagcgggccg cgcagaaagc tgatgtcctg accactggag 180ctggtaaccc agtaggagac aaacttaatg ttattacagt agggccccgt gggccccttc 240ttgttcagga tgtggttttc actgatgaaa tggctcattt tgaccgagag agaattcctg 300agagagttgt gcatgctaaa ggagcagggg cctttggcta ctttgaggtc acacatgaca 360ttaccaaata ctccaaggca aaggtatttg agcatattgg aaagaagact cccatcgcag 420ttcggttctc cactgttgct ggagaatcgg gttcagctga cacagttcgg gaccctcgtg 480ggtttgcagt gaaattttac acagaagatg gtaactggga tctcgttgga aataacaccc 540ccattttctt catcagggat cccatattgt ttccatcttt tatccacagc caaaagagaa 600atcctcagac acatctgaag gatccggaca tggtctggga cttctggagc ctacgtcctg 660agtctctgca tcaggtttct ttcttgttca gtgatcgggg gattccagat ggacatcgcc 720acatgaatgg atatggatca catactttca agctggttaa tgcaaatggg gaggcagttt 780attgcaaatt ccattataag actgaccagg gcatcaaaaa cctttctgtt gaagatgcgg 840cgagactttc ccaggaagat cctgactatg gcatccggga tctttttaac gccattgcca 900caggaaagta cccctcctgg actttttaca tccaggtcat gacatttaat caggcagaaa 960cttttccatt taatccattc gatctcacca aggtttggcc tcacaaggac taccctctca 1020tcccagttgg taaactggtc ttaaaccgga atccagttaa ttactttgct gaggttgaac 1080agatagcctt cgacccaagc aacatgccac ctggcattga ggccagtcct gacaaaatgc 1140ttcagggccg cctttttgcc tatcctgaca ctcaccgcca tcgcctggga cccaattatc 1200ttcatatacc tgtgaactgt ccctaccgtg ctcgagtggc caactaccag cgtgacggcc 1260cgatgtgcat gcaggacaat cagggtggtg ctccaaatta ctaccccaac agctttggtg 1320ctccggaaca acagccttct gccctggagc acagcatcca atattctgga gaagtgcgga 1380gattcaacac tgccaatgat gataacgtta ctcaggtgcg ggcattctat gtgaacgtgc 1440tgaatgagga acagaggaaa cgtctgtgtg agaacattgc cggccacctg aaggatgcac 1500aaattttcat ccagaagaaa gcggtcaaga acttcactga ggtccaccct gactacggga 1560gccacatcca ggctcttctg gacaagtaca atgctgagaa gcctaagaat gcgattcaca 1620cctttgtgca gtccggatct cacttggcgg caagggagaa ggcaaatctg tgaggccggg 1680gccctgcacc tgtgcagcga agcttagcgt tcatccgtgt aacccgctca tcactggatg 1740aagattctcc tgtgctagat gtgcaaatgc aagctagtgg cttcaaaata gagaatccca 1800ctttctatag cagattgtgt aacaatttta atgctatttc cccaggggaa aatgaaggtt 1860aggatttaac agtcatttaa aaaaaaaatt tgttttgacg gatgattgga ttattcattt 1920aaaatgatta gaaggcaagt ttctagctag aaatatgatt ttatttgaca aaatttgttg 1980aaattatgta tgtttacata tcacctcatg gcctattata ttaaaatatg gctataaata 2040tataaaaaga aaagataaag atgatctact cagaaatttt tatttttcta aggttctcat 2100aggaaaagta catttaatac agcagtgtca tcagaagata acttgagcac cgtcatggct 2160taatgtttat tcctgataat aattgatcaa attcattttt ttcactggag ttacattaat 2220gttaattcag cactgatttc acaacagatc aatttgtaat tgcttacatt tttacaataa 2280ataatctgta cgtaagaaca 230051262DNAHomo sapiens 5actctcgcga gatccctact ggctataaag gcagcgcccc ggagagctct tgcgcgtctt 60gttcttgcct ggtgtcggtg gttagtttct gcgacttgtg ttgggactgg tgagtgtggg 120cagtgcggcc cctgcggagt gaggcgcggc gcgcccttct tgcctgttgc ctcttcctcc 180tcctgtccgg ggcccgcccg cgctcgggtg ggggtgctgt gatgcgtgag gcagccgggg 240gaggcccgga gtccgagact gcttgagcgc tgcgcacacc cctctcgtgg gccccccacg 300taggtgcggg aacctggttg aaccccaagc tgataggaag atgtcttcag gaaatgctaa 360aattgggcac cctgccccca acttcaaagc cacagctgtt atgccagatg gtcagtttaa 420agatatcagc ctgtctgact acaaaggaaa atatgttgtg ttcttctttt accctcttga 480cttcaccttt gtgtgcccca cggagatcat tgctttcagt gatagggcag aagaatttaa 540gaaactcaac tgccaagtga ttggtgcttc tgtggattct cacttctgtc atctagcatg 600ggtcaataca cctaagaaac aaggaggact gggacccatg aacattcctt tggtatcaga 660cccgaagcgc accattgctc aggattatgg ggtcttaaag gctgatgaag gcatctcgtt 720caggggcctt tttatcattg atgataaggg tattcttcgg cagatcactg taaatgacct 780ccctgttggc cgctctgtgg atgagacttt gagactagtt caggccttcc agttcactga 840caaacatggg gaagtgtgcc cagctggctg gaaacctggc agtgatacca tcaagcctga 900tgtccaaaag agcaaagaat atttctccaa gcagaagtga gcgctgggct gttttagtgc 960caggctgcgg tgggcagcca tgagaacaaa acctcttctg tatttttttt ttccattagt 1020aaaacacaag acttcagatt cagccgaatt gtggtgtctt acaaggcagg cctttcctac 1080agggggtgga gagaccagcc tttcttcctt tggtaggaat ggcctgagtt ggcgttgtgg 1140gcaggctact ggtttgtatg atgtattagt agagcaaccc attaatcttt tgtagtttgt 1200attaaacttg aactgagacc ttgatgagtc tttaaaaaaa aaaaaaaaaa aaaaaaaaaa 1260aa 12626827DNAHomo sapiens 6gcagtggagg cggcccaggc ccgccttccg cagggtgtcg ccgctgtgcc gctagcggtg 60ccccgcctgc tgcggtggca ccagccagga ggcggagtgg aagtggccgt ggggcgggta 120tgggactagc tggcgtgtgc gccctgagac gctcagcggg ctatatactc gtcggtgggg 180ccggcggtca gtctgcggca gcggcagcaa gacggtgcag tgaaggagag tgggcgtctg 240gcggggtccg cagtttcagc agagccgctg cagccatggc cccaatcaag gtgggagatg 300ccatcccagc agtggaggtg tttgaagggg agccagggaa caaggtgaac ctggcagagc 360tgttcaaggg caagaagggt gtgctgtttg gagttcctgg ggccttcacc cctggatgtt 420ccaaggttcg gctcctggct gatcccactg gggcctttgg gaaggagaca gacttattac 480tagatgattc gctggtgtcc atctttggga atcgacgtct caagaggttc tccatggtgg 540tacaggatgg catagtgaag gccctgaatg tggaaccaga tggcacaggc ctcacctgca 600gcctggcacc caatatcatc tcacagctct gaggccctgg gccagattac ttcctccacc 660cctccctatc tcacctgccc agccctgtgc tggggccctg caattggaat gttggccaga 720tttctgcaat aaacacttgt ggtttgcggc caaaaaaaaa aaaaaaaaaa aaaaaaaaaa 780aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaa 82771593DNAHomo sapiens 7gcggtgccct tgcggcgcag ctggggtcgc ggccctgctc cccgcgcttt cttaaggccc 60gcgggcggcg caggagcggc actcgtggct gtggtggctt cggcagcggc ttcagcagat 120cggcggcatc agcggtagca ccagcactag cagcatgttg agccgggcag tgtgcggcac 180cagcaggcag ctggctccgg ttttggggta tctgggctcc aggcagaagc acagcctccc 240cgacctgccc tacgactacg gcgccctgga acctcacatc aacgcgcaga tcatgcagct 300gcaccacagc aagcaccacg cggcctacgt gaacaacctg aacgtcaccg aggagaagta 360ccaggaggcg ttggccaagg gagatgttac agcccagata gctcttcagc ctgcactgaa 420gttcaatggt ggtggtcata tcaatcatag cattttctgg acaaacctca gccctaacgg 480tggtggagaa cccaaagggg agttgctgga agccatcaaa cgtgactttg gttcctttga 540caagtttaag gagaagctga cggctgcatc tgttggtgtc caaggctcag gttggggttg 600gcttggtttc aataaggaac ggggacactt acaaattgct gcttgtccaa atcaggatcc 660actgcaagga acaacaggcc ttattccact gctggggatt gatgtgtggg agcacgctta 720ctaccttcag tataaaaatg tcaggcctga ttatctaaaa gctatttgga atgtaatcaa 780ctgggagaat gtaactgaaa gatacatggc ttgcaaaaag taaaccacga tcgttatgct 840gagtatgtta agctctttat gactgttttt gtagtggtat agagtactgc agaatacagt 900aagctgctct attgtagcat ttcttgatgt tgcttagtca cttatttcat aaacaactta 960atgttctgaa taatttctta ctaaacattt tgttattggg caagtgattg aaaatagtaa 1020atgctttgtg tgattgaatc tgattggaca ttttcttcag agagctaaat tacaattgtc 1080atttataaaa ccatcaaaaa tattccatcc atatactttg gggacttgta gggatgcctt 1140tctagtccta ttctattgca gttatagaaa atctagtctt ttgccccagt tacttaaaaa 1200taaaatatta acactttccc aagggaaaca ctcggctttc tatagaaaat tgcacttttt 1260gtcgagtaat cctctgcagt gatacttctg gtagatgtca cccagtggtt tttgttaggt 1320caaatgttcc tgtatagttt ttgcaaatag agctgtatac tgtttaaatg tagcaggtga 1380actgaactgg ggtttgctca cctgcacagt aaaggcaaac ttcaacagca aaactgcaaa 1440aaggtggttt ttgcagtagg agaaaggagg atgtttattt gcagggcgcc aagcaaggag 1500aattgggcag ctcatgcttg agacccaatc tccatgatga cctacaagct agagtattta 1560aaggcagtgg taaatttcag gaaagcagaa gtt 159383174DNAHomo sapiens 8tcctcctggg tcttgcctag cggcgggcgc atgcttagtc accgtgaggc tgcgcttgcc 60cggggcccgc gcccccctac cccggggacc gcccccgggc cgcccgcccc acttggcgcg 120ccacttccgc gtgcatggcc ctgctgcccc gagccctgag cgccggcgcg ggaccgagct 180ggcggcgggc ggcgcgcgcc ttccgaggct tcctgctgct tctgcccgag cccgcggccc 240tcacgcgcgc cctctcccgt gccatggcct gcaggcagga gccgcagccg cagggcccgc 300cgcccgctgc tggcgccgtg gcctcctatg actacctggt gatcgggggc ggctcgggcg 360ggctggccag cgcgcgcagg gcggccgagc tgggtgccag ggccgccgtg gtggagagcc 420acaagctggg tggcacttgc gtgaatgttg gatgtgtacc caaaaaggta atgtggaaca 480cagctgtcca ctctgaattc atgcatgatc atgctgatta tggctttcca agttgtgagg 540gtaaattcaa ttggcgtgtt attaaggaaa agcgggatgc ctatgtgagc cgcctgaatg 600ccatctatca aaacaatctc accaagtccc atatagaaat catccgtggc catgcagcct 660tcacgagtga tcccaagccc acaatagagg tcagtgggaa aaagtacacc gccccacaca 720tcctgatcgc cacaggtggt atgccctcca cccctcatga gagccagatc cccggtgcca 780gcttaggaat aaccagcgat ggattttttc agctggaaga attgcccggc cgcagcgtca 840ttgttggtgc aggttacatt gctgtggaga tggcagggat cctgtcagcc ctgggttcta 900agacatcact gatgatacgg catgataagg tacttagaag ttttgattca atgatcagca 960ccaactgcac ggaggagctg gagaacgctg gcgtggaggt gctgaagttc tcccaggtca 1020aggaggttaa aaagactttg tcgggcttgg aagtcagcat ggttactgca gttcccggta 1080ggctaccagt catgaccatg attccagatg ttgactgcct gctctgggcc attgggcggg 1140tcccgaatac caaggacctg agtttaaaca aactggggat tcaaaccgat gacaagggtc 1200atatcatcgt agacgaattc cagaatacca acgtcaaagg catctatgca gttggggatg 1260tatgtggaaa agctcttctt actccagttg caatagctgc tggccgaaaa cttgcccatc 1320gactttttga atataaggaa gattccaaat tagattataa caacatccca actgtggtct 1380tcagccaccc ccctattggg acagtgggac tcacggaaga tgaagccatt cataaatatg 1440gaatagaaaa tgtgaagacc tattcaacga gctttacccc gatgtatcac gcagttacca 1500aaaggaaaac aaaatgtgtg atgaaaatgg tctgtgctaa caaggaagaa aaggtggttg 1560ggatccatat gcagggactt gggtgtgatg aaatgctgca gggttttgct gttgcagtga 1620agatgggagc aacgaaggca gactttgaca acacagtcgc cattcaccct acctcttcag 1680aagagctggt cacacttcgt tgagaaccag gagacacgtg tggcgggcag tgggacccat 1740agatcttctg aaatgaaaca aataatcaca ttgacttact gtttgagttt tatgtatttc 1800tttattttaa tcaggatctt ctgatagtgg aaatttttag tacataatag aacttattta 1860tggagttaga aatttgtagt gttatccagg attgattttc atttgatcac atctcacagt 1920aattaatatt ttcaagtttt ttttttatta acagctctgt gctagttttt tttttctgtt 1980ttagcctcat cccaaatata aagctttgtg aagtacaatt aacttaatgt acttgaatga 2040atagaacttg ctactttttt tttttttttt tttgagacag agttttgctc tcattgccca 2100ggctggagtg cggtggtgct atttcagctc accacaacct ctgcctcctg ggttcaagtg 2160attctcctgc cttagcctcc cgaatagctg gaattacagg cacgcaccac catgcctgac 2220taattttgta tttttagtag acatggggtt tctccatgtt ggtcaggctg gtctcaaact 2280cccaccttca ggtgatccgc ccacctcggc ctcctgaggt gctgagatta caggcgtgag 2340ccactgtgcc agcttgctaa ttttcacaga agttgatggc aattcttcac atgtaaacag 2400tgccagtgca cagaaccttt atatattttt tgaagccagt actgtgctct gcatataaca 2460aagctgcttc aaggatgaga cctttttcta aaagcatgta atgtgagaag ccggcctgcc 2520ttattttctt ttttcttttt taatgattaa aaatagtttg tggcaaggca cggtggctca 2580ggcctgtaat tctagcactt tgggaggccg aggcaggagg attacttgag cctacaagtt 2640tgaggccagc atgcacagca tagcaagact gcatctctac agagagtaaa aaaaattacc 2700cgagtgtggt gatgtgcatc tgtaatctca gctacttggg aggctgaggt gagaggatca 2760cttgagcttg ggtgaggtga ggctgcagtg agtcctgatc atgctgctgc actcaatctt 2820ggacaacaga gcaagaccct gtctcaaaaa aaaaaaaaaa aaatatatat atatatatat 2880attattttta tgaggtgaag tgcatcaaac ttgggaaaga tttgaggagg ctgggaacct 2940cctggaaaac cactccttga agaaagatat gagagacatt tagaagtgat tcctgctttc 3000agaaggaggt ggattcaaat acatcaaaag tcccttcctc tgctaagtgt ttatagttca 3060atgaataatt tcaatatttg tatgtgttct tgtcatttta tttttttctg aaaaacttcc 3120aaaaatttga aaataaaatt acagcctttt cttcttataa aaaaaaaaaa aaaa 317491661DNAHomo sapiens 9attgcattcc tgggcattgc taactagtga agtataccag atggaaatgt cttcgaagct 60gtccctttaa aactcgagca agctaccagg caaactccgc ctccagggag gttccttatt 120aaataggagc caactggctg ggtcggggct caatacccca agcaatacct gcaactgagg 180attcttcccg gggagaccgc agcccatcgg catggctcaa gagtttgtga actgcaaaat 240ccagcctggg aaggtggttg tgttcatcaa gcccacctgc ccgtactgca ggagggccca 300agagatcctc agtcaattgc ccatcaaaca agggcttctg gaatttgtcg atatcacagc 360caccaaccac actaacgaga ttcaagatta tttgcaacag ctcacgggag caagaacggt 420gcctcgagtc tttattggta aagattgtat aggcggatgc agtgatctag tctctttgca 480acagagtggg gaactgctga cgcggctaaa gcagattgga gctctgcagt aaccacagaa 540caggccccat gctgacgtcc ctcctcaaga gctggatggc attgcaaatg atgacagcac 600ttctggtgga tgaatttggg ggcacaaaca gcttttttcc tcttttggct cagtatttaa 660aagtggacca acttgctctt aatcacaggg ccaagaaggt tgacgggcca tcttggtttt 720cttctggatg tgctctttgg ttttcagaag actgtgacaa gttctggccc aggattcgct 780cactgaccct caattgtcct ctttggcatg cgtttcttac tgttctccat gtgtcggcat 840gtctctacct ctaagccagt gtttttcaac tatgtttatc cagactcctt ctccacaatg 900atgaatccac agttggttat ctgctactgc ccattagcta aaatcatttt gctgcttgac 960tttatggagt ttgtattatg aaatcagtgg gtattttgaa tgtgttcttt ctaactacat 1020gcatctctcc actcaactcc accccatccc atcccacctt gaaaatcact gctctgaacc 1080agtgttctcc accttgtcct ccacagatct cataggaaat gttcaacaat tctgtgaaag 1140gtcacaggac ccaattggag aaatcatatg aaaagcatag ttggtcttgg tgtcatatgg 1200atcagaggca caagtgcaga ggctgtggtc atgcggaaca ctctgttatt taagatggct 1260atccagataa tcctgaacac tgtgtattta ttttatttag actaccagca aagattaaag 1320catgaaatgt aaaacatctg ataaaactta cagcccccta caccaagagt gtatctgtga 1380aagagctcct acactttgaa aacttaagaa tcccttatca tgaagtttgc ctgttctaga 1440attgtaagat tgttaatttc cttcaatctc tagtgacaac acttaatttc ttttctaata 1500aaaaaaacct atagatgatt cagtgatttt tgtccaattc atttgcatgt tctcaagaca 1560ttaaggaatg ttatgcgaaa tacactaact taaaactgtg tttatatttg gccctgccat 1620tataaataaa gacacgtgct gctgtcaaaa aaaaaaaaaa a 1661101039DNAHomo sapiens 10gctcgtccgc tccctccccc gcgccgtgca cgtcttggtt cgggccgggc ataaaaggct 60tcgcggccca gggctcactt ggcgctgaga acgcgggtcc acgcgtgtga tcgtccgtgc 120gtctagcctt tgcccacgca gctttcagtc atggcctccg gtaacgcgcg catcggaaag 180ccagcccctg acttcaaggc cacagcggtg gttgatggcg ccttcaaaga ggtgaagctg 240tcggactaca aagggaagta cgtggtcctc tttttctacc ctctggactt cacttttgtg 300tgccccaccg agatcatcgc gttcagcaac cgtgcagagg

acttccgcaa gctgggctgt 360gaagtgctgg gcgtctcggt ggactctcag ttcacccacc tggcttggat caacaccccc 420cggaaagagg gaggcttggg ccccctgaac atccccctgc ttgctgacgt gaccagacgc 480ttgtctgagg attacggcgt gctgaaaaca gatgagggca ttgcctacag gggcctcttt 540atcatcgatg gcaagggtgt ccttcgccag atcactgtta atgatttgcc tgtgggacgc 600tccgtggatg aggctctgcg gctggtccag gccttccagt acacagacga gcatggggaa 660gtttgtcccg ctggctggaa gcctggcagt gacacgatta agcccaacgt ggatgacagc 720aaggaatatt tctccaaaca caattaggct ggctaacgga tagtgagctt gtgcccctgc 780ctaggtgcct gtgctgggtg tccacctgtg cccccacctg ggtgccctat gctgacccag 840gaaaggccag acctgcccct ccaaactcca cagtatggga ccctggaggg ctaggccaag 900gccttctcat gcctccacct agaagctgaa tagtgacgcc ctcccccaag cccacccagc 960cgcacacagg cctagaggta accaataaag tattagggaa aggtgtgaaa aaaaaaaaaa 1020aaaaaaaaaa aaaaaaaaa 1039112799DNAHomo sapiens 11acgtgtgcgg gagggaagca ggaagtgact gcgggagtgg agccggcgag agagtggcag 60cgggggctga tggaagtgca gtgggggctg gagagggcac cctactgtat ccagcatgct 120ccaaggccac agctctgtgt tccaggcctt gctggggacc ttcttcacct gggggatgac 180agcagctggg gcagctctcg tgttcgtatt ctctagtgga cagaggcgga tcttagatgg 240aagtcttggc tttgctgcag gggtcatgtt ggcagcttcc tattggtctc ttctggcccc 300agcagttgag atggccacgt cctctggggg cttcggtgcc tttgccttct tccctgtggc 360tgttggcttc acccttggag cggcttttgt ctacttggct gacctcctga tgcctcactt 420gggtgcagca gaagaccccc agacgaccct ggcactgaac ttcggctcta cgttgatgaa 480gaagaagtct gatcctgagg gtcccgcgct gctcttccct gagagtgaac tttccatccg 540gataggtaga gctgggcttc tttcagacaa gagtgagaat ggtgaggcat atcagagaaa 600gaaggcggca gccactggcc ttccagaggg tcctgctgtc cctgtgcctt ctcgagggaa 660tctggcacag cccggcggca gcagctggag gaggatcgca ctgctcatct tggccatcac 720tatacacaac gttccagagg gtctcgctgt tggagttgga tttggggcta tagaaaagac 780ggcatctgct acctttgaga gtgccaggaa tttggccatt ggaatcggga tccagaattt 840ccccgagggc ctggctgtca gccttccctt gcgaggggca ggcttctcca cctggagagc 900tttctggtat gggcagctga gcggcatggt ggagcccctg gccggggtct ttggtgcctt 960tgccgtggtg ctggctgagc ccatcctgcc ctacgctctg gcctttgctg ccggtgccat 1020ggtctacgtg gtcatggacg acatcatccc cgaagcccag atcagtggta atgggaaact 1080ggcatcctgg gcctccatcc tgggatttgt agtgatgatg tcactggacg ttggcctggg 1140ctagggctga gacgcttcgg accccgggaa aggccatacg aagaaacagc agtggttggc 1200ttctatggga caacaagctt ctttcttcac attaaaactt ttttccttcc tctcttcttc 1260atctcattat cctgattgac tctgattata atagaaccat ttttactttg ctttgaggga 1320gatttttgat ttaatgggga attttaaggt gtcatggaaa tacagattct ttgttttggc 1380cactgaatgg actctctctt cagtgggatt atcaaggaac ttcagatcag ggaaatctcc 1440acttcgggac cttctatctg cctcccaact cctcaaggtc acctatagaa gcgagctacc 1500aaaagacgtc tcctaagcat tttggtggcc tagtgactca gggcagagtg gccagcacac 1560ctctcatccg cccctcctgc tccatcactg ctgagcctct ccccatctag aatgttggaa 1620ctggagcatc ataaagatag caagctacct tccaaggccg agccagccca gagaggagca 1680tgtcttcctt tacctccccc taaggagata ctacatggga gggggacaca gaaaaaggga 1740aggaaattgg ctagtctggc tttttttttt ttttttttta aaggcaaaga ttgacattat 1800tgaaggaaag gggatgagga caactgtgaa ctcacagtga gccctgtgga aagaagagac 1860agacagagtg tgggtttgtt cggaggcctc tgctgtcaat ggattccagg agcaaggcca 1920tttgtcgcgc tttccaaatt tcttaggcat ttattttgat aagtttatag ccatcatgtt 1980tctaagagac ttggagacac cagcaaactg ctagaactca aactcttcaa ttactcaaag 2040aaggagccat ttcagttaac tcaagtgaat gaaagagttt tggaatctgc tgtgggtcct 2100tccctgttga ccatttggta acttataatc tgacaaaaac tcttgagctg caacaggcct 2160tgccagaggg ctcaggatgg gaaaggaaga aggggatagg aaaagaagag gtaattttac 2220atttcccctt taaagtaaat tttagccaac tcatcattct gaaatgtccc tataaagaat 2280gagtcgaact agaccagaag ccagcctact ccttcttaca tagcttctcc aacaggggta 2340gcaatgacct gtccacttca aacacagata aggcctgcca tcctcattgg ttaaaggcac 2400acgtgagact ttcagtgggc tctgctgaga aggaaggcag cccaggagtc aggtatgcag 2460gcattgcatt gtcagtgtct gctctcagag tttacacatt caattgcttc caagggtgaa 2520tctcctgctc tgtgaatgct atcagacccc aaaggccaac cttgggctgg gtctatgtac 2580gttcttccga agcactgatg atcaaaattg aagacacatt cagaggtttg attggttgag 2640attaactggt gtggtggttg gtgtatgtat gttttatttt tatgtctttg tatgtagttc 2700tacataatgc aaattgtgct ttctgatgga caagacctca taactgtgat taatatcaat 2760aaaaagggga tgttgtggat gaaaaaaaaa aaaaaaaaa 279912981DNAHomo sapiens 12gtttggggcc agagtgggcg aggcgcggag gtctggccta taaagtagtc gcggagacgg 60ggtgctggtt tgcgtcgtag tctcctgcag cgtctggggt ttccgttgca gtcctcggaa 120ccaggacctc ggcgtggcct agcgagttat ggcgacgaag gccgtgtgcg tgctgaaggg 180cgacggccca gtgcagggca tcatcaattt cgagcagaag gaaagtaatg gaccagtgaa 240ggtgtgggga agcattaaag gactgactga aggcctgcat ggattccatg ttcatgagtt 300tggagataat acagcaggct gtaccagtgc aggtcctcac tttaatcctc tatccagaaa 360acacggtggg ccaaaggatg aagagaggca tgttggagac ttgggcaatg tgactgctga 420caaagatggt gtggccgatg tgtctattga agattctgtg atctcactct caggagacca 480ttgcatcatt ggccgcacac tggtggtcca tgaaaaagca gatgacttgg gcaaaggtgg 540aaatgaagaa agtacaaaga caggaaacgc tggaagtcgt ttggcttgtg gtgtaattgg 600gatcgcccaa taaacattcc cttggatgta gtctgaggcc ccttaactca tctgttatcc 660tgctagctgt agaaatgtat cctgataaac attaaacact gtaatcttaa aagtgtaatt 720gtgtgacttt ttcagagttg ctttaaagta cctgtagtga gaaactgatt tatgatcact 780tggaagattt gtatagtttt ataaaactca gttaaaatgt ctgtttcaat gacctgtatt 840ttgccagact taaatcacag atgggtatta aacttgtcag aatttctttg tcattcaagc 900ctgtgaataa aaaccctgta tggcacttat tatgaggcta ttaaaagaat ccaaattcaa 960actaaaaaaa aaaaaaaaaa a 98113508DNAHomo sapiens 13tttggtgctt tggatccatt tccatcggtc cttacagccg ctcgtcagac tccagcagcc 60aagatggtga agcagatcga gagcaagact gcttttcagg aagccttgga cgctgcaggt 120gataaacttg tagtagttga cttctcagcc acgtggtgtg ggccttgcaa aatgatcaag 180cctttctttc attccctctc tgaaaagtat tccaacgtga tattccttga agtagatgtg 240gatgactgtc aggatgttgc ttcagagtgt gaagtcaaat gcatgccaac attccagttt 300tttaagaagg gacaaaaggt gggtgaattt tctggagcca ataaggaaaa gcttgaagcc 360accattaatg aattagtcta atcatgtttt ctgaaaatat aaccagccat tggctattta 420aaacttgtaa tttttttaat ttacaaaaat ataaaatatg aagacataaa cccagttgcc 480atctgcgtga caataaaaca ttaatgct 508141591DNAHomo sapiens 14ccctgcgtct ctgcccgccc cgtggcgccc gagtgcactg aagatggcgg ctgctgtagg 60acggttgctc cgagcgtcgg ttgcccgaca tgtgagtgcc attccttggg gcatttctgc 120cactgcagcc ctcaggcctg ctgcatgtgg aagaacgagc ttgacaaatt tattgtgttc 180tggttccagt caagcaaaat tattcagcac cagttcctca tgccatgcac ctgctgtcac 240ccagcatgca ccctatttta agggtacagc cgttgtcaat ggagagttca aagacctaag 300ccttgatgac tttaagggga aatatttggt gcttttcttc tatcctttgg atttcacctt 360tgtgtgtcct acagaaattg ttgcttttag tgacaaagct aacgaatttc acgacgtgaa 420ctgtgaagtt gtcgcagtct cagtggattc ccactttagc catcttgcct ggataaatac 480accaaggaag aatggtggtt tgggccacat gaacatcgca ctcttgtcag acttaactaa 540gcagatttcc cgagactacg gtgtgctgtt agaaggttct ggtcttgcac taagaggtct 600cttcataatt gaccccaatg gagtcatcaa gcatttgagc gtcaacgatc tcccagtggg 660ccgaagcgtg gaagaaaccc tccgcttggt gaaggcgttc cagtatgtag aaacacatgg 720agaagtctgc ccagcgaact ggacaccgga ttctcctacg atcaagccaa gtccagctgc 780ttccaaagag tactttcaga aggtaaatca gtagatcacc catgtgtatc tgcaccttct 840caactgagag aagaaccaca gttgaaacct gcttttatca ttttcaagat ggttatttgt 900agaaggcaag gaaccaatta tgcttgtatt cataagtatt actctaaatg ttttgttttt 960gtaattctgg ctaagacctt ttaaacatgg ttagttgcta gtacaaggaa tcctttattg 1020gtaacatctt ggtggctggc tagctagttt ctacagaaca taatttgcct ctatagaagg 1080ctattcttag atcatgtctc aatggaaaca ctcttctttc ttagccttac ttgaatcttg 1140cctataataa agtagagcaa cacacattga aagcttctga tcaacggtcc tgaaattttc 1200atcttgaatg tctttgtatt aaactgaatt ttcttttaag ctaacaaaga tcataatttt 1260caatgattag ccgtgtaact cctgcaatga atgtttatgt gattgaagca aatgtgaatc 1320gtattatttt aaaaagtggc agagtgactt aactgatcat gcatgatccc tcatccctga 1380aattgagttt atgtagtcat tttacttatt ttattcatta gctaactttg tctatgtata 1440tttctagata ttgattagtg taatcgatta taaaggatat ttatcaaatc cagggattgc 1500attttgaaat tataattatt ttctttgctg aagtattcat tgtaaaacat acaaaataaa 1560catattttaa aacatttgca ttttaccacc a 1591151246DNAHomo sapiens 15gtctttgccc tcgcgacgcc gccacctccg gaacaagcca tggtggcggc gacggtggca 60gcggcgtggc tgctcctgtg ggctgcggcc tgcgcgcagc aggagcagga cttctacgac 120ttcaaggcgg tcaacatccg gggcaaactg gtgtcgctgg agaagtaccg cggatcggtg 180tccctggtgg tgaatgtggc cagcgagtgc ggcttcacag accagcacta ccgagccctg 240cagcagctgc agcgagacct gggcccccac cactttaacg tgctcgcctt cccctgcaac 300cagtttggcc aacaggagcc tgacagcaac aaggagattg agagctttgc ccgccgcacc 360tacagtgtct cattccccat gtttagcaag attgcagtca ccggtactgg tgcccatcct 420gccttcaagt acctggccca gacttctggg aaggagccca cctggaactt ctggaagtac 480ctagtagccc cagatggaaa ggtggtaggg gcttgggacc caactgtgtc agtggaggag 540gtcagacccc agatcacagc gctcgtgagg aagctcatcc tactgaagcg agaagactta 600taaccaccgc gtctcctcct ccaccacctc atcccgccca cctgtgtggg gctgaccaat 660gcaaactcaa atggtgcttc aaagggagag acccactgac tctccttcct ttactcttat 720gccattggtc ccatcattct tgtgggggaa aaattctagt attttgatta tttgaatctt 780acagcaacaa ataggaactc ctggccaatg agagctcttg accagtgaat caccagccga 840tacgaacgtc ttgccaacaa aaatgtgtgg caaatagaag tatatcaagc aataatctcc 900cacccaaggc ttctgtaaac tgggaccaat gattacctca tagggctgtt gtgaggatta 960ggatgaaata cctgtgaaag tgcctaggca gtgccagcca aataggaggc attcaatgaa 1020cattttttgc atataaacca aaaaataact tgttatcaat aaaaacttgc atccaacatg 1080aatttccagc cgatgataat ccaggccaaa ggtttagttg ttgttatttc ctctgtatta 1140ttttcttcat tacaaaagaa atgcaagttc attgtaacaa tccaaacaat acctcacgat 1200ataaaataaa aatgaaagta tcctcctcaa aaaaaaaaaa aaaaaa 124616942DNAHomo sapiens 16gagcgctctg gagggcgtgg ccgtgggaaa ggaggcgcgg aaagccgacg cgcgtccatt 60ggtcggctgg acgaggggag gagccgctgg ctcccagccc cgccgcgatg agcctcggcc 120gcctttgccg cctactgaag ccggcgctgc tctgtggggc tctggccgcg cctggcctgg 180ccgggaccat gtgcgcgtcc cgggacgact ggcgctgtgc gcgctccatg cacgagtttt 240ccgccaagga catcgacggg cacatggtta acctggacaa gtaccggggc ttcgtgtgca 300tcgtcaccaa cgtggcctcc cagtgaggca agaccgaagt aaactacact cagctcgtcg 360acctgcacgc ccgatacgct gagtgtggtt tgcggatcct ggccttcccg tgtaaccagt 420tcgggaagca ggagccaggg agtaacgaag agatcaaaga gttcgccgcg ggctacaacg 480tcaaattcga tatgttcagc aagatctgcg tgaacgggga cgacgcccac ccgctgtgga 540agtggatgaa gatccaaccc aagggcaagg gcatcctggg aaatgccatc aagtggaact 600tcaccaagtt cctcatcgac aagaacggct gcgtggtgaa gcgctacgga cccatggagg 660agcccctggt gatagagaag gacctgcccc actatttcta gctccacaag tgtgtggccc 720cgcccgagcc cctgcccacg cccttggagc cttccaccgg cactcatgac ggcctgcctg 780caaacctgct ggtggggcag acccgaaaat ccagcgtgca ccccgccgga ggaaggtccc 840atggcctgct gggcttggct cggcgccccc acccctggct accttgtggg aataaacaga 900caaattagcc tgctggaaaa aaaaaaaaaa aaaaaaaaaa aa 942171342DNAHomo sapiens 17gaggaagtga cgacaggcgt gcccttgaca ggcagggagg gctaggctgt gcatccctcc 60gctcgcattg cagggagatg gctcagcgac ttcttctgag gaggttcctg gcctctgtca 120tctccaggaa gccctctcag ggtcagtggc cacccctcac ttccagagcc ctgcagaccc 180cacaatgcag tcctggtggc ctgactgtaa cacccaaccc agcccggaca atatacacca 240cgaggatctc cttgacaacc tttaatatcc aggatggacc tgactttcaa gaccgagtgg 300tcaacagtga gacaccagtg gttgtggatt tccacgcaca gtggtgtgga ccctgcaaga 360tcctggggcc gaggttagag aagatggtgg ccaagcagca cgggaaggtg gtgatggcca 420aggtggatat tgatgaccac acagacctcg ccattgagta tgaggtgtca gcggtgccca 480ctgtgctggc catgaagaat ggggacgtgg tggacaagtt tgtgggcatc aaggatgagg 540atcagttgga ggccttcctg aagaagctga ttggctgaca agcagggatg agtcctggtt 600cccttgcccg cgtgggaccc caatagaact cagcccttcc atgccagccc ttcctgctgc 660ctccctcctg tctggctcct ggggcccatg cttagagccc aggctccagc cctgagtgct 720tccgagctgg cggactgccc aggggccatc agaggatggt ggtgctgctg ctgatccggg 780gaccgctgtc ttccctccca tacgcctttc atccctcctt ctagggccta tggcagttct 840cccaggatgt gtggcgagag cctgggccag cccacagcgt tcctagtcag gcagccacac 900cttggtcctc atcttggtcc cttccaatct gaaacctcgt gcctggctcg tctgccacct 960acatttctct ttccagctgc tgttttgtaa aaagaaaaag aaaaaagaag cccaaactag 1020tgagagtaat atctaattat ctcatttttt gtaggtctgt gataaagaac ttagtcatcc 1080cttccacctc ctactgtgaa gaacagaccc tgggtcccac actgaaatcc cctctagtca 1140cccattccca ccccccaggg agctgcctcc caggcagggg gtgcagaaaa tgattgatgg 1200gctggggaac cctggagagc ctcgactccg gaagtctcaa ggtgcctcct cctctcctta 1260gctggcccgt tggttttctg agcagggggc tgaactgtga acaagtcaga caaataaagc 1320aagggtctgc accatcaaaa aa 134218921DNAHomo sapiens 18gcggcgctcg cgccaaggga cgtgtttctg cgctcgcgtg gtcatggagg cgctgccgct 60gctagccgcg acaactccgg accacggccg ccaccgaagg ctgcttctgc tgccgctact 120gctgttcctg ctgccggctg gagctgtgca gggctgggag acagaggaga ggccccggac 180tcgcgaagag gagtgccact tctacgcggg tggacaagtg tacccgggag aggcatcccg 240ggtatcggtc gccgaccact ccctgcacct aagcaaagcg aagatttcca agccagcgcc 300ctactgggaa ggaacagctg tgatcgatgg agaatttaag gagctgaagt taactgatta 360tcgtgggaaa tacttggttt tcttcttcta cccacttgat ttcacatttg tgtgtccaac 420tgaaattatc gcttttggcg acagacttga agaattcaga tctataaata ctgaagtggt 480agcatgctct gttgattcac agtttaccca tttggcctgg attaataccc ctcgaagaca 540aggaggactt gggccaataa ggattccact tctttcagat ttgacccatc agatctcaaa 600ggactatggt gtatacctag aggactcagg ccacactctt agaggtctct tcattattga 660tgacaaagga atcctaagac aaattactct gaatgatctt cctgtgggta gatcagtgga 720tgagacacta cgtttggttc aagcattcca gtacactgac aaacacggag aagtctgccc 780tgctggctgg aaacctggta gtgaaacaat aatcccagat ccagctggaa agctgaagta 840tttcgataaa ctgaattgag aaatacttct tcaagttatg atgcttgaaa gttctcaata 900aagttcacgg tttcattacc a 921191200DNAHomo sapiens 19cagttaaaag gaggcgcctg ctggcctccc cttacagtgc ttgttcgggg cgctccgctg 60gcttcttgga caattgcgcc atgtgtgctg ctcggctagc ggcggcggcg gcggcggccc 120agtcggtgta tgccttctcg gcgcgcccgc tggccggcgg ggagcctgtg agcctgggct 180ccctgcgggg caaggtacta cttatcgaga atgtggcgtc cctctgaggc accacggtcc 240gggactacac ccagatgaac gagctgcagc ggcgcctcgg accccggggc ctggtggtgc 300tcggcttccc gtgcaaccag tttgggcatc aggtgcgccg ggcggagcgg ggcggggcgg 360gggcggacgt gcagtagtgg ctgggggcgc cggcggtgtg ctggtgggtg ccgtcggctc 420catgcgcgga gagtctggct actctctcgt ttcctttctg ttgctcgtag ctgctgaaat 480tcctctccgc ccttgggatt gcgcatggag ggcaaaatcc cggtgactca tagaaaatct 540cccttgtttg tggttagaac gtttctctcc tcctcttgac cccgggttct agctgccctt 600ctctcctgta ggagaacgcc aagaacgaag agattctgaa ttccctcaag tacgtccggc 660ctggtggtgg gttcgagccc aacttcatgc tcttcgagaa gtgcgaggtg aacggtgcgg 720gggcgcaccc tctcttcgcc ttcctgcggg aggccctgcc agctcccagc gacgacgcca 780ccgcgcttat gaccgacccc aagctcatca cctggtctcc ggtgtgtcgc aacgatgttg 840cctggaactt tgagaagttc ctggtgggcc ctgacggtgt gcccctacgc aggtacagcc 900gccgcttcca gaccattgac atcgagcctg acatcgaagc cctgctgtct caagggccca 960gctgtgccta gggcgcccct cctaccccgg ctgcttggca gttgcagtgc tgctgtctcg 1020ggggggtttt catctatgag ggtgtttcct ctaaacctac gagggaggaa cacctgatct 1080tacagaaaat accacctcga gatgggtgct ggtcctgttg atcccagtct ctgccagacc 1140aaggcgagtt tccccactaa taaagtgccg ggtgtcagca gaaaaaaaaa aaaaaaaaaa 1200201274DNAHomo sapiens 20gcttctgtct ggcggcggca gcatggcggc gggggcggct gaggcagctg tagcggccgt 60ggaggaggtc ggctcagccg ggcagtttga ggagctgctg cgcctcaaag ccaagtccct 120ccttgtggtc catttctggg caccatgggc tccacagtgt gcacagatga acgaagttat 180ggcagagtta gctaaagaac tccctcaagt ttcatttgtg aagttggaag ctgaaggtgt 240tcctgaagta tctgaaaaat atgaaattag ctctgttccc acttttctgt ttttcaagaa 300ttctcagaaa atcgaccgat tagatggtgc acatgcccca gagttgacca aaaaagttca 360gcgacatgca tctagtggct ccttcctacc cagcgctaat gaacatctta aagaagatct 420caaccttcgc ttgaagaaat tgactcatgc tgccccctgc atgctgttta tgaaaggaac 480tcctcaagaa ccacgctgtg gtttcagcaa gcagatggtg gaaattcttc acaaacataa 540tattcagttt agcagttttg atatcttctc agatgaagag gttcgacagg gactcaaagc 600ctattccagt tggcctacct atcctcagct ctatgtttct ggagagctca taggaggact 660tgatataatt aaggagctag aagcatctga agaactagat acaatttgtc ccaaagctcc 720caaattagag gaaaggctca aagtgctgac aaataaagct tctgtgatgc tctttatgaa 780aggaaacaaa caggaagcaa aatgtggatt cagcaaacaa attctggaaa tactaaatag 840tactggtgtt gaatatgaaa cattcgatat attggaggat gaagaagttc ggcaaggatt 900aaaagcttac tcaaattggc caacataccc tcagctgtat gtgaaagggg agctggtggg 960aggattggat attgtgaagg aactgaaaga aaatggtgaa ttgctgccta tactgagagg 1020agaaaattaa taaatcttaa acttggtgcc caactattgt aagaaatatt taattacatt 1080gggagcagtt catgatttag tcctcagaaa tggactagga atagaaaatt cctgctttct 1140cagttacatg ttttgtgtat ttcacaatgt cgtgctaaat aaatgtatgt tacatttttt 1200tcccaccaaa aatagaatgc aataaacatc ttcaaattat taacgaaaaa aaaaaaaaaa 1260aaaaaaaaaa aaaa 127421710DNAHomo sapiens 21gctcgtccgc tccctccccc gcgccgtgca cgtcttggtt cgggccgggc ataaaaggct 60tcgcggccca gggctcactt ggcgctgaga acgcgggtcc acgcgtgtga tcgtccgtgc 120gtctagcctt tgcccacgca gctttcagtc atggcctccg gtaacgcgcg catcggaaag 180ccagcccctg acttcaaggc cacagcggtg gttgatggcg ccttcaaaga ggtgaagctg 240tcggactaca aagggaagta cgtggtcctc tttttctacc ctctggactt cacttttgtg 300tgccccaccg agatcatcgc gttcagcaac cgtgcagagg acttccgcaa gctgggctgt 360gaagtgctgg gcgtctcggt ggactctcag ttcacccacc tggcttggta tgagcagggg 420ccaaagaggg aggttgcagc taagctcaca ccctcaggtc ctagcagtgt ggcttcgtgg 480ccattgctca acctctggaa cctgcgtttc cccatcgtga aaataatgga aacattgccg 540cccaagtctt taaggatgat gacagtaatt agcatttgac aactagttgc ctggtatata 600gagttgcaga tgcaactcag atgcaactct atctactcta tgtacttagt tcccaggagg 660gaggctgtgc tgccctattt catgaagatg gaaactccag ttcaccgaag 710221715DNAHomo sapiens 22gaaccaaccg gttgcttgct gtcccagcgg cgccccctca tcaccgtcgc catgcccgga 60ggtctgcttc tcggggacgt ggctcccaac tttgaggcca ataccaccgt cggccgcatc 120cgtttccacg actttctggg agactcatgg ggcattctct tctcccaccc tcgggacttt 180accccagtgt gcaccacaga gcttggcaga gctgcaaagc tggcaccaga atttgccaag 240aggaatgtta agttgattgc cctttcaata gacagtgttg

aggaccatct tgcctggagc 300aaggatatca atgcttacaa ttgtgaagag cccacagaaa agttaccttt tcccatcatc 360gatgatagga atcgggagct tgccatcctg ttgggcatgc tggatccagc agagaaggat 420gaaaagggca tgcctgtgac agctcgtgtg gtgtttgttt ttggtcctga taagaagctg 480aagctgtcta tcctctaccc agctaccact ggcaggaact ttgatgagat tctcagggta 540gtcatctctc tccagctgac agcagaaaaa agggttgcca ccccagttga ttggaaggat 600ggggatagtg tgatggtcct tccaaccatc cctgaagaag aagccaaaaa acttttcccg 660aaaggagtct tcaccaaaga gctcccatct ggcaagaaat acctccgcta cacaccccag 720ccttaagtct cttggagaag ctggtgctgt gagccagagg atgtcagctg ccaattgtgt 780tttcctgcag caattccata aacacatcct ggtgtcatca cagccaaggt ttttaggttg 840ctataccaat ggcttattaa atgaaaatgg cactaaaagt ttcttgagat tctttatact 900ctctgccttc agcaatcaat tccattcata catcagcact ctgctggttc tgtttgaaat 960atgttctgta tttaaaactc aaatcttgtt ggatctctgc agggcttgtg accaatgaag 1020tcatatttgt tgatggttga caaagcttgc ttcactccat cagagaatga ctatcaattt 1080ttttttaact gtcctatcac gtcctctcct gtcacccatt ttgaagagtg gcagaacttg 1140aagttcaact tcctctgtaa atatccaagt ataaagccca ggaacttcta gaataaccca 1200gatgcgcttt aatttttttt aatatgtttt gatcacagaa cttctagaat aacccagatg 1260ctctttcata ttcttttaat acatcttgat cacagctggg ggaaaaaaag ctttttaatt 1320ctataccttc ctagtagata agtgaagagc agggaaagag acctttaaat attttgctat 1380aaaaaaattt gtgataagtt tctatcaaaa tggggagatt gcagaaaagg cttcccttgg 1440ctcccaagga ggtgtagcag gtgtgagcaa tattagtgcc atgtgccttt cacacagggt 1500ttgcatttat cagtctgttt tccgatgatg tgtacatgaa agagtacacc atgtgaagag 1560aagagagaat gattgaaaat gttttagtat agaactcttc ttgcagtggg ttgctatttt 1620ctagatttta ctttttaggg aacaaaataa aatcctttgt taaaactggg aaaaaaaaaa 1680aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaa 1715231182DNAHomo sapiens 23aatgagggcc tccagggggc gggtcggact gccgcgggcc ggggagcgct ctgggtggcc 60agctgtgggc ccgggccgtc gtgggctccg gcttgcgtgc ggagatgagc gggtccctcg 120gccgagctgc ggcggctctg ctccgctggg ggcgcggcgc gggcggcggt ggcctttggg 180gtccgggcgt gcgggcggcg ggctcgggcg cgggcggcgg cggctcggcg gagcagttgg 240acgcgctggt gaagaaggac aaggtggtgg tcttcctcaa ggggacgccg gagcagcccc 300agtgcggctt cagcaacgcc gtggtgcaga tcctgcggct gcacggcgtc cgcgattacg 360cggcctacaa cgtgctggac gacccggagc tccgacaagg cattaaagac tattccaact 420ggcccaccat cccgcaagtg tacctcaatg gcgagtttgt agggggctgt gacattcttc 480tgcagatgca ccagaatggg gacttggtgg aagaactgaa aaagctgggg atccactccg 540cccttttaga tgaaaagaaa gaccaagact ccaagtgagg gcggccaagt cctcgctgag 600cagagaggga gccgttcatg tcagagactc actgccagaa aagccttacc cattttggtt 660ttcactattg agaccgcaac tgcttgcact gatcattttg gttcgtgagc agttggtgat 720tttagttggt ctggtgttcg ggctaagaat attttattgt ggacttaatt acaaccactg 780cactgtaatg attcaatgct gtattatgat attgctgtaa acaaaattca ttcttatatt 840gtcacttatt ctttgcctga ttcagaagtt aaataggagc tttggaatca ttattcatga 900cccctctgca aatgtgtcag tctccaaaga gagtatctcc ccccaaattt tgtgtagctt 960cttttgttat ggaaaatggt gaacaaaaaa agaaactgtg ataactgggg cgttgttttt 1020taaaataaac tccagcacag ggatgctgtg catgcctgag ttgattccga agtgcatatg 1080tctgtaagga tttggagtgc ctgcagtgtt ttatgtgtgg gaagtaaggg tgagtctcat 1140attcttctat taaatttgcc acaagaattg caaaaaaaaa aa 1182241170DNAHomo sapiens 24aaggctatta ttaccaccac tgagtggctt aaataatcct gtcaacagca atcgcccatt 60tccaaagcca tggtgaaaca tctctgtgct aatttctttt gttttgtttc ctaatttttt 120ttttttggca ggtggtggga aataatcttt gtcttctttg gagtaaacct tcaacaccgg 180attttttctt ttaattatgg atgtaaaccc caatatcccc ataatttaca ttgggtctcg 240accaattgcc taattataag aggatatatt taggctctta tttcatccac acaaaaactt 300gtgtaacagg tagttggaaa catctgaggc accactttga ttctgttttg gatggtcatg 360ttttttctcc tccgtttccc cagcatgtct gccaccatcc tcatgcactg cttccaagtg 420cctgggagcc tttatgagcg tccctaaacc taaaagaatc cagaggcggg gctcggatga 480accctcgaga taagcaagtg agccgcttct cccctctaaa ggatgtttac acgtgggtgg 540cactcgctgg aatccagcgc tcgggcagcc ctgggaggac gcgctcagct gcgaggagga 600tggagagcaa tacatcatca tctttggaga atttagcgac ggcgcctgtg aaccagatcc 660aagaaacaat ttctgataat tgtgtggtga ttttctcaaa aacatcctgt tcttactgta 720caatggcaaa aaagcttttc catgacatga atgttaacta taaagtggtg gaactggacc 780tgcttgaata tggaaaccag ttccaagatg ctctttacaa aatgactggt gaaagaactg 840ttccaagaat atttgtcaat ggtactttta ttggaggtgc aactgacact cataggcttc 900acaaagaagg aaaattgctc ccactagttc atcagtgtta tttaaaaaaa agtaagagga 960aagaatttca gtgatgttta tactaataag tttgctagta cagtgtcagt tatttaaagt 1020ggtaatgccc gataatgtct tttaaatgtt tgaggatgtt ttaaatacat gcattgtctt 1080cacgaagaag atgtaaaaat aatgaacaat aaattgcggt ggaaacctaa aaaaaaaaaa 1140aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 11702518DNAArtificial sequenceoligonucleotide sens 25gggagctgag ggcaagtc 182619DNAArtificial sequenceoligonucleotide antisens 26gagacccttg cagccaatc 192719DNAArtificial sequenceoligonucleotide sens 27caaccagttt gggcatcag 192819DNAArtificial sequenceoligonucleotide antisens 28gttcacctcg cacttctcg 192918DNAArtificial sequenceoligonucleotide sens 29gctggtttgg agcaggag 183024DNAArtificial sequenceoligonucleotide antisens 30ccaaagatga tgatgtattg ctct 243118DNAArtificial sequenceoligonucleotide sens 31cgcagttcgg ttctccac 183218DNAArtificial sequenceoligonucleotide antisens 32gggtcccgaa ctgtgtca 183321DNAArtificial sequenceoligonucleotide sens 33cactgacaaa catggggaag t 213421DNAArtificial sequenceoligonucleotide antisens 34tttgctcttt tggacatcag g 213518DNAArtificial sequenceoligonucleotide sens 35cacccctgga tgttccaa 183622DNAArtificial sequenceoligonucleotide antisens 36ggacaccagc gaatcatcta gt 223720DNAArtificial sequenceoligonucleotide sens 37tccactgcaa ggaacaacag 203819DNAArtificial sequenceoligonucleotide antisens 38taagcgtgct cccacacat 193921DNAArtificial sequenceoligonucleotide sens 39tgccagctta ggaataacca g 214018DNAArtificial sequenceoligonucleotide antisens 40cctgcaccaa caatgacg 184120DNAArtificial sequenceoligonucleotide sens 41ggcttctgga atttgtcgat 204221DNAArtificial sequenceoligonucleotide antisens 42tgcatccgcc tatacaatct t 214321DNAArtificial sequenceoligonucleotide sens 43gccttccagt acacagacga g 214421DNAArtificial sequenceoligonucleotide antisens 44gttgggctta atcgtgtcac t 214518DNAArtificial sequenceoligonucleotide sens 45tcctggctga tcccactg 184620DNAArtificial sequenceoligonucleotide antisens 46atgccatcct gtaccaccat 204720DNAArtificial sequenceoligonucleotide sens 47gcatcatcaa tttcgagcag 204820DNAArtificial sequenceoligonucleotide antisens 48caggccttca gtcagtcctt 204918DNAArtificial sequenceoligonucleotide sens 49ttacagccgc tcgtcaga 185021DNAArtificial sequenceoligonucleotide antisens 50ggcttcctga aaagcagtct t 215120DNAArtificial sequenceoligonucleotide sens 51ctggacaccg gattctccta 205225DNAArtificial sequenceoligonucleotide antisens 52gggtgatcta ctgatttacc ttctg 255320DNAArtificial sequenceoligonucleotide sens 53ccatcctgcc ttcaagtacc 205420DNAArtificial sequenceoligonucleotide antisens 54ttccatctgg ggctactagg 205518DNAArtificial sequenceoligonucleotide sens 55tacggaccca tggaggag 185622DNAArtificial sequenceoligonucleotide antisens 56ccacacactt gtggagctag aa 225720DNAArtificial sequenceoligonucleotide sens 57gagacaccag tggttgtgga 205819DNAArtificial sequenceoligonucleotide antisens 58gcttggccac catcttctc 195920DNAArtificial sequenceoligonucleotide sens 59gcacctaagc aaagcgaaga 206021DNAArtificial sequenceoligonucleotide antisens 60aaattctcca tcgatcacag c 216121DNAArtificial sequenceoligonucleotide sens 61cccttgtttg tggttagaac g 216221DNAArtificial sequenceoligonucleotide antisens 62gagagaaggg cagctagaac c 216319DNAArtificial sequenceoligonucleotide sens 63tcctcaagaa ccacgctgt 196427DNAArtificial sequenceoligonucleotide antisens 64tgagaagata tcaaaactgc taaactg 276526DNAArtificial sequenceoligonucleotide sens 65gcaactcaga tgcaactcta tctact 266623DNAArtificial sequenceoligonucleotide antisens 66tgaactggag tttccatctt cat 236724DNAArtificial sequenceoligonucleotide sens 67caatagacag tgttgaggac catc 246820DNAArtificial sequenceoligonucleotide antisens 68tttctgtggg ctcttcacaa 206920DNAArtificial sequenceoligonucleotide sens 69gtgataactg gggcgttgtt 207018DNAArtificial sequenceoligonucleotide antisens 70actcaggcat gcacagca 187118DNAArtificial sequenceoligonucleotide sens 71gtggcactcg ctggaatc 187222DNAArtificial sequenceoligonucleotide antisens 72cgtcgctaaa ttctccaaag at 22731236DNAHomo sapiens 73actctcgcga gatccctact ggctataaag gcagcgcccc ggagagctct tgcgcgtctt 60gttcttgcct ggtgtcggtg gttagtttct gcgacttgtg ttgggactgg tgagtgtggg 120cagtgcggcc cctgcggagt gaggcgcggc gcgcccttct tgcctgttgc ctcttcctcc 180tcctgtccgg ggcccgcccg cgctcgggtg ggggtgctgt gatgcgtgag gcagccgggg 240gaggcccgga gtccgagact gcttgagcgc tgcgcacacc cctctcgtgg gccccccacg 300tagctgatag gaagatgtct tcaggaaatg ctaaaattgg gcaccctgcc cccaacttca 360aagccacagc tgttatgcca gatggtcagt ttaaagatat cagcctgtct gactacaaag 420gaaaatatgt tgtgttcttc ttttaccctc ttgacttcac ctttgtgtgc cccacggaga 480tcattgcttt cagtgatagg gcagaagaat ttaagaaact caactgccaa gtgattggtg 540cttctgtgga ttctcacttc tgtcatctag catgggtcaa tacacctaag aaacaaggag 600gactgggacc catgaacatt cctttggtat cagacccgaa gcgcaccatt gctcaggatt 660atggggtctt aaaggctgat gaaggcatct cgttcagggg cctttttatc attgatgata 720agggtattct tcggcagatc actgtaaatg acctccctgt tggccgctct gtggatgaga 780ctttgagact agttcaggcc ttccagttca ctgacaaaca tggggaagtg tgcccagctg 840gctggaaacc tggcagtgat accatcaagc ctgatgtcca aaagagcaaa gaatatttct 900ccaagcagaa gtgagcgctg ggctgtttta gtgccaggct gcggtgggca gccatgagaa 960caaaacctct tctgtatttt ttttttccat tagtaaaaca caagacttca gattcagccg 1020aattgtggtg tcttacaagg caggcctttc ctacaggggg tggagagacc agcctttctt 1080cctttggtag gaatggcctg agttggcgtt gtgggcaggc tactggtttg tatgatgtat 1140tagtagagca acccattaat cttttgtagt ttgtattaaa cttgaactga gaccttgatg 1200agtctttaaa aaaaaaaaaa aaaaaaaaaa aaaaaa 1236741042DNAHomo sapiens 74actctcgcga gatccctact ggctataaag gcagcgcccc ggagagctct tgcgcgtctt 60gttcttgcct ggtgtcggtg gttagtttct gcgacttgtg ttgggactgc tgataggaag 120atgtcttcag gaaatgctaa aattgggcac cctgccccca acttcaaagc cacagctgtt 180atgccagatg gtcagtttaa agatatcagc ctgtctgact acaaaggaaa atatgttgtg 240ttcttctttt accctcttga cttcaccttt gtgtgcccca cggagatcat tgctttcagt 300gatagggcag aagaatttaa gaaactcaac tgccaagtga ttggtgcttc tgtggattct 360cacttctgtc atctagcatg ggtcaataca cctaagaaac aaggaggact gggacccatg 420aacattcctt tggtatcaga cccgaagcgc accattgctc aggattatgg ggtcttaaag 480gctgatgaag gcatctcgtt caggggcctt tttatcattg atgataaggg tattcttcgg 540cagatcactg taaatgacct ccctgttggc cgctctgtgg atgagacttt gagactagtt 600caggccttcc agttcactga caaacatggg gaagtgtgcc cagctggctg gaaacctggc 660agtgatacca tcaagcctga tgtccaaaag agcaaagaat atttctccaa gcagaagtga 720gcgctgggct gttttagtgc caggctgcgg tgggcagcca tgagaacaaa acctcttctg 780tatttttttt ttccattagt aaaacacaag acttcagatt cagccgaatt gtggtgtctt 840acaaggcagg cctttcctac agggggtgga gagaccagcc tttcttcctt tggtaggaat 900ggcctgagtt ggcgttgtgg gcaggctact ggtttgtatg atgtattagt agagcaaccc 960attaatcttt tgtagtttgt attaaacttg aactgagacc ttgatgagtc tttaaaaaaa 1020aaaaaaaaaa aaaaaaaaaa aa 1042751035DNAHomo sapiens 75gcggtgccct tgcggcgcag ctggggtcgc ggccctgctc cccgcgcttt cttaaggccc 60gcgggcggcg caggagcggc actcgtggct gtggtggctt cggcagcggc ttcagcagat 120cggcggcatc agcggtagca ccagcactag cagcatgttg agccgggcag tgtgcggcac 180cagcaggcag ctggctccgg ttttggggta tctgggctcc aggcagaagc acagcctccc 240cgacctgccc tacgactacg gcgccctgga acctcacatc aacgcgcaga tcatgcagct 300gcaccacagc aagcaccacg cggcctacgt gaacaacctg aacgtcaccg aggagaagta 360ccaggaggcg ttggccaagg gagatgttac agcccagata gctcttcagc ctgcactgaa 420gttcaatggt ggtggtcata tcaatcatag cattttctgg acaaacctca gccctaacgg 480tggtggagaa cccaaagggg agttgctgga agccatcaaa cgtgactttg gttcctttga 540caagtttaag gagaagctga cggctgcatc tgttggtgtc caaggctcag gttggggttg 600gcttggtttc aataaggaac ggggacactt acaaattgct gcttgtccaa atcaggatcc 660actgcaagga acaacaggcc ttattccact gctggggatt gatgtgtggg agcacgctta 720ctaccttcag tataaaaatg tcaggcctga ttatctaaaa gctatttgga atgtaatcaa 780ctgggagaat gtaactgaaa gatacatggc ttgcaaaaag taaaccacga tcgttatgct 840gatcataccc taatgatccc agcaagataa tgtcctgtct tctaagatgt gcatcaagcc 900tggtacatac tgaaaaccct ataaggtcct ggataatttt tgtttgatta ttcattgaag 960aaacatttat tttccaattg tgtgaagttt ttgactgtta ataaaagaat ctgtcaacca 1020tcaaaaaaaa aaaaa 103576918DNAHomo sapiens 76gcggtgccct tgcggcgcag ctggggtcgc ggccctgctc cccgcgcttt cttaaggccc 60gcgggcggcg caggagcggc actcgtggct gtggtggctt cggcagcggc ttcagcagat 120cggcggcatc agcggtagca ccagcactag cagcatgttg agccgggcag tgtgcggcac 180cagcaggcag ctggctccgg ttttggggta tctgggctcc aggcagaagc acagcctccc 240cgacctgccc tacgactacg gcgccctgga acctcacatc aacgcgcaga tcatgcagct 300gcaccacagc aagcaccacg cggcctacgt gaacaacctg aacgtcaccg aggagaagta 360ccaggaggcg ttggccaagg gggagttgct ggaagccatc aaacgtgact ttggttcctt 420tgacaagttt aaggagaagc tgacggctgc atctgttggt gtccaaggct caggttgggg 480ttggcttggt ttcaataagg aacggggaca cttacaaatt gctgcttgtc caaatcagga 540tccactgcaa ggaacaacag gccttattcc actgctgggg attgatgtgt gggagcacgc 600ttactacctt cagtataaaa atgtcaggcc tgattatcta aaagctattt ggaatgtaat 660caactgggag aatgtaactg aaagatacat ggcttgcaaa aagtaaacca cgatcgttat 720gctgatcata ccctaatgat cccagcaaga taatgtcctg tcttctaaga tgtgcatcaa 780gcctggtaca tactgaaaac cctataaggt cctggataat ttttgtttga ttattcattg 840aagaaacatt tattttccaa ttgtgtgaag tttttgactg ttaataaaag aatctgtcaa 900ccatcaaaaa aaaaaaaa 918773087DNAHomo sapiens 77tcctcctggg tcttgcctag cggcgggcgc atgcttagtc accgtgaggc tgcgcttgcc 60cggggcccgc gcccccctac cccggggacc gcccccgggc cgcccgcccc acttggcgcg 120ccacttccgc gtgcatggcc ctgctgcccc gagccctgag cgccggcgcg ggaccgagct 180ggcggcgggc ggcgcgcgcc ttccgaggct tcctgctgct tctgcccgag cccgcggccc 240tcacgcgcgc cctctcccgt gccatggcct gcaggcagga gccgcagccg cagggcccgc 300cgcccgctgc tggcgccgtg gcctcctatg actacctggt gatcgggggc ggctcgggcg 360ggctggccag cgcgcgcagg gcggccgagc tgggtgccag ggccgccgtg gtggagagcc 420acaagctggg tggcacttgc gtgaatgttg gatgtgtacc caaaaaggta atgtggaaca 480cagctgtcca ctctgaattc atgcatgatc atgctgatta tggctttcca agttgtgagg 540gtaaattcaa ttggcgtgtt attaaggaaa agcgggatgc ctatgtgagc cgcctgaatg 600ccatctatca aaacaatctc accaagtccc atatagaaat catccgtggc catgcagcct 660tcacgagtga tcccaagccc acaatagagg tcagtgggaa aaagtacacc gccccacaca 720tcctgatcgc cacaggtggt atgccctcca cccctcatga gagccagatc cccggtgcca 780gcttaggaat aaccagcgat ggattttttc agctggaaga attgcccggc cgcagcgtca 840ttgttggtgc aggttacatt gctgtggaga tggcagggat cctgtcagcc ctgggttcta 900agacatcact gatgatacgg catgataagg tcaaggaggt taaaaagact ttgtcgggct 960tggaagtcag catggttact gcagttcccg gtaggctacc agtcatgacc atgattccag 1020atgttgactg cctgctctgg gccattgggc gggtcccgaa taccaaggac ctgagtttaa 1080acaaactggg gattcaaacc gatgacaagg gtcatatcat cgtagacgaa ttccagaata 1140ccaacgtcaa aggcatctat gcagttgggg atgtatgtgg aaaagctctt cttactccag 1200ttgcaatagc tgctggccga aaacttgccc atcgactttt tgaatataag gaagattcca 1260aattagatta taacaacatc ccaactgtgg tcttcagcca cccccctatt gggacagtgg 1320gactcacgga agatgaagcc attcataaat atggaataga aaatgtgaag acctattcaa 1380cgagctttac cccgatgtat

cacgcagtta ccaaaaggaa aacaaaatgt gtgatgaaaa 1440tggtctgtgc taacaaggaa gaaaaggtgg ttgggatcca tatgcaggga cttgggtgtg 1500atgaaatgct gcagggtttt gctgttgcag tgaagatggg agcaacgaag gcagactttg 1560acaacacagt cgccattcac cctacctctt cagaagagct ggtcacactt cgttgagaac 1620caggagacac gtgtggcggg cagtgggacc catagatctt ctgaaatgaa acaaataatc 1680acattgactt actgtttgag ttttatgtat ttctttattt taatcaggat cttctgatag 1740tggaaatttt tagtacataa tagaacttat ttatggagtt agaaatttgt agtgttatcc 1800aggattgatt ttcatttgat cacatctcac agtaattaat attttcaagt ttttttttta 1860ttaacagctc tgtgctagtt ttttttttct gttttagcct catcccaaat ataaagcttt 1920gtgaagtaca attaacttaa tgtacttgaa tgaatagaac ttgctacttt tttttttttt 1980ttttttgaga cagagttttg ctctcattgc ccaggctgga gtgcggtggt gctatttcag 2040ctcaccacaa cctctgcctc ctgggttcaa gtgattctcc tgccttagcc tcccgaatag 2100ctggaattac aggcacgcac caccatgcct gactaatttt gtatttttag tagacatggg 2160gtttctccat gttggtcagg ctggtctcaa actcccacct tcaggtgatc cgcccacctc 2220ggcctcctga ggtgctgaga ttacaggcgt gagccactgt gccagcttgc taattttcac 2280agaagttgat ggcaattctt cacatgtaaa cagtgccagt gcacagaacc tttatatatt 2340ttttgaagcc agtactgtgc tctgcatata acaaagctgc ttcaaggatg agaccttttt 2400ctaaaagcat gtaatgtgag aagccggcct gccttatttt cttttttctt ttttaatgat 2460taaaaatagt ttgtggcaag gcacggtggc tcaggcctgt aattctagca ctttgggagg 2520ccgaggcagg aggattactt gagcctacaa gtttgaggcc agcatgcaca gcatagcaag 2580actgcatctc tacagagagt aaaaaaaatt acccgagtgt ggtgatgtgc atctgtaatc 2640tcagctactt gggaggctga ggtgagagga tcacttgagc ttgggtgagg tgaggctgca 2700gtgagtcctg atcatgctgc tgcactcaat cttggacaac agagcaagac cctgtctcaa 2760aaaaaaaaaa aaaaaatata tatatatata tatattattt ttatgaggtg aagtgcatca 2820aacttgggaa agatttgagg aggctgggaa cctcctggaa aaccactcct tgaagaaaga 2880tatgagagac atttagaagt gattcctgct ttcagaagga ggtggattca aatacatcaa 2940aagtcccttc ctctgctaag tgtttatagt tcaatgaata atttcaatat ttgtatgtgt 3000tcttgtcatt ttattttttt ctgaaaaact tccaaaaatt tgaaaataaa attacagcct 3060tttcttctta taaaaaaaaa aaaaaaa 3087783014DNAHomo sapiens 78tcctcctggg tcttgcctag cggcgggcgc atgcttagtc accgtgaggc tgcgcttgcc 60cggggcccgc gcccccctac cccggggacc gcccccgggc cgcccgcccc acttggcgcg 120ccacttccgc gtgcatggcc ctgctgcccc gagccctgag cgccggcgcg ggaccgagct 180ggcggcgggc ggcgcgcgcc ttccgaggct tcctgctgct tctgcccgag cccgcggccc 240tcacgcgcgc cctctcccgt gccatggcct gcaggcagga gccgcagccg cagggcccgc 300cgcccgctgc tggcgccgtg gcctcctatg actacctggt gatcgggggc ggctcgggcg 360ggctggccag cgcgcgcagg gcggccgagc tgggtgccag ggccgccgtg gtggagagcc 420acaagctggg tggcacttgc gtgaatgttg gatgtgtacc caaaaaggta atgtggaaca 480cagctgtcca ctctgaattc atgcatgatc atgctgatta tggctttcca agttgtgagg 540gtaaattcaa ttggcgtgtt attaaggaaa agcgggatgc ctatgtgagc cgcctgaatg 600ccatctatca aaacaatctc accaagtccc atatagaaat catccgtggc catgcagcct 660tcacgagtga tcccaagccc acaatagagg tcagtgggaa aaagtacacc gccccacaca 720tcctgatcgc cacaggtggt atgccctcca cccctcatga gagccagatc cccggtgcca 780gcttaggaat aaccagcgat ggattttttc agctggaaga attgcccggc cgcagcgtca 840ttgttggtgc aggttacatt gctgtggaga tggcagggat cctgtcagcc ctgggttcta 900agacatcact gatgatacgg catgataagg tacttagaag ttttgattca atgatcagca 960ccaactgcac ggaggagctg gagaacgctg gcgtggaggt gctgaagttc tcccagggga 1020ttcaaaccga tgacaagggt catatcatcg tagacgaatt ccagaatacc aacgtcaaag 1080gcatctatgc agttggggat gtatgtggaa aagctcttct tactccagtt gcaatagctg 1140ctggccgaaa acttgcccat cgactttttg aatataagga agattccaaa ttagattata 1200acaacatccc aactgtggtc ttcagccacc cccctattgg gacagtggga ctcacggaag 1260ataagccatt cataaatatg gaatagaaaa tgtgaagacc tattcaacga gctttacccc 1320gatgtatcac gcagttacca aaaggaaaac aaaatgtgtg atgaaaatgg tctgtgctaa 1380caaggaagaa aaggtggttg ggatccatat gcagggactt gggtgtgatg aaatgctgca 1440gggttttgct gttgcagtga agatgggagc aacgaaggca gactttgaca acacagtcgc 1500cattcaccct acctcttcag aagagctggt cacacttcgt tgagaaccag gagacacgtg 1560tggcgggcag tgggacccat agatcttctg aaatgaaaca aataatcaca ttgacttact 1620gtttgagttt tatgtatttc tttattttaa tcaggatctt ctgatagtgg aaatttttag 1680tacataatag aacttattta tggagttaga aatttgtagt gttatccagg attgattttc 1740atttgatcac atctcacagt aattaatatt ttcaagtttt ttttttatta acagctctgt 1800gctagttttt tttttctgtt ttagcctcat cccaaatata aagctttgtg aagtacaatt 1860aacttaatgt acttgaatga atagaacttg ctactttttt tttttttttt tttgagacag 1920agttttgctc tcattgccca ggctggagtg cggtggtgct atttcagctc accacaacct 1980ctgcctcctg ggttcaagtg attctcctgc cttagcctcc cgaatagctg gaattacagg 2040cacgcaccac catgcctgac taattttgta tttttagtag acatggggtt tctccatgtt 2100ggtcaggctg gtctcaaact cccaccttca ggtgatccgc ccacctcggc ctcctgaggt 2160gctgagatta caggcgtgag ccactgtgcc agcttgctaa ttttcacaga agttgatggc 2220aattcttcac atgtaaacag tgccagtgca cagaaccttt atatattttt tgaagccagt 2280actgtgctct gcatataaca aagctgcttc aaggatgaga cctttttcta aaagcatgta 2340atgtgagaag ccggcctgcc ttattttctt ttttcttttt taatgattaa aaatagtttg 2400tggcaaggca cggtggctca ggcctgtaat tctagcactt tgggaggccg aggcaggagg 2460attacttgag cctacaagtt tgaggccagc atgcacagca tagcaagact gcatctctac 2520agagagtaaa aaaaattacc cgagtgtggt gatgtgcatc tgtaatctca gctacttggg 2580aggctgaggt gagaggatca cttgagcttg ggtgaggtga ggctgcagtg agtcctgatc 2640atgctgctgc actcaatctt ggacaacaga gcaagaccct gtctcaaaaa aaaaaaaaaa 2700aaatatatat atatatatat attattttta tgaggtgaag tgcatcaaac ttgggaaaga 2760tttgaggagg ctgggaacct cctggaaaac cactccttga agaaagatat gagagacatt 2820tagaagtgat tcctgctttc agaaggaggt ggattcaaat acatcaaaag tcccttcctc 2880tgctaagtgt ttatagttca atgaataatt tcaatatttg tatgtgttct tgtcatttta 2940tttttttctg aaaaacttcc aaaaatttga aaataaaatt acagcctttt cttcttataa 3000aaaaaaaaaa aaaa 3014792928DNAHomo sapiens 79tcctcctggg tcttgcctag cggcgggcgc atgcttagtc accgtgaggc tgcgcttgcc 60cggggcccgc gcccccctac cccggggacc gcccccgggc cgcccgcccc acttggcgcg 120ccacttccgc gtgcatggcc ctgctgcccc gagccctgag cgccggcgcg ggaccgagct 180ggcggcgggc ggcgcgcgcc ttccgaggct tcctgctgct tctgcccgag cccgcggccc 240tcacgcgcgc cctctcccgt gccatggcct gcaggcagga gccgcagccg cagggcccgc 300cgcccgctgc tggcgccgtg gcctcctatg actacctggt gatcgggggc ggctcgggcg 360ggctggccag cgcgcgcagg gcggccgagc tgggtgccag ggccgccgtg gtggagagcc 420acaagctggg tggcacttgc gtgaatgttg gatgtgtacc caaaaaggta atgtggaaca 480cagctgtcca ctctgaattc atgcatgatc atgctgatta tggctttcca agttgtgagg 540gtaaattcaa ttggcgtgtt attaaggaaa agcgggatgc ctatgtgagc cgcctgaatg 600ccatctatca aaacaatctc accaagtccc atatagaaat catccgtggc catgcagcct 660tcacgagtga tcccaagccc acaatagagg tcagtgggaa aaagtacacc gccccacaca 720tcctgatcgc cacaggtggt atgccctcca cccctcatga gagccagatc cccggtgcca 780gcttaggaat aaccagcgat ggattttttc agctggaaga attgcccggc cgcagcgtca 840ttgttggtgc aggttacatt gctgtggaga tggcagggat cctgtcagcc ctgggttcta 900agacatcact gatgatacgg catgataagg ggattcaaac cgatgacaag ggtcatatca 960tcgtagacga attccagaat accaacgtca aaggcatcta tgcagttggg gatgtatgtg 1020gaaaagctct tcttactcca gttgcaatag ctgctggccg aaaacttgcc catcgacttt 1080ttgaatataa ggaagattcc aaattagatt ataacaacat cccaactgtg gtcttcagcc 1140acccccctat tgggacagtg ggactcacgg aagatgaagc cattcataaa tatggaatag 1200aaaatgtgaa gacctattca acgagcttta ccccgatgta tcacgcagtt accaaaagga 1260aaacaaaatg tgtgatgaaa atggtctgtg ctaacaagga agaaaaggtg gttgggatcc 1320atatgcaggg acttgggtgt gatgaaatgc tgcagggttt tgctgttgca gtgaagatgg 1380gagcaacgaa ggcagacttt gacaacacag tcgccattca ccctacctct tcagaagagc 1440tggtcacact tcgttgagaa ccaggagaca cgtgtggcgg gcagtgggac ccatagatct 1500tctgaaatga aacaaataat cacattgact tactgtttga gttttatgta tttctttatt 1560ttaatcagga tcttctgata gtggaaattt ttagtacata atagaactta tttatggagt 1620tagaaatttg tagtgttatc caggattgat tttcatttga tcacatctca cagtaattaa 1680tattttcaag tttttttttt attaacagct ctgtgctagt tttttttttc tgttttagcc 1740tcatcccaaa tataaagctt tgtgaagtac aattaactta atgtacttga atgaatagaa 1800cttgctactt tttttttttt tttttttgag acagagtttt gctctcattg cccaggctgg 1860agtgcggtgg tgctatttca gctcaccaca acctctgcct cctgggttca agtgattctc 1920ctgccttagc ctcccgaata gctggaatta caggcacgca ccaccatgcc tgactaattt 1980tgtattttta gtagacatgg ggtttctcca tgttggtcag gctggtctca aactcccacc 2040ttcaggtgat ccgcccacct cggcctcctg aggtgctgag attacaggcg tgagccactg 2100tgccagcttg ctaattttca cagaagttga tggcaattct tcacatgtaa acagtgccag 2160tgcacagaac ctttatatat tttttgaagc cagtactgtg ctctgcatat aacaaagctg 2220cttcaaggat gagacctttt tctaaaagca tgtaatgtga gaagccggcc tgccttattt 2280tcttttttct tttttaatga ttaaaaatag tttgtggcaa ggcacggtgg ctcaggcctg 2340taattctagc actttgggag gccgaggcag gaggattact tgagcctaca agtttgaggc 2400cagcatgcac agcatagcaa gactgcatct ctacagagag taaaaaaaat tacccgagtg 2460tggtgatgtg catctgtaat ctcagctact tgggaggctg aggtgagagg atcacttgag 2520cttgggtgag gtgaggctgc agtgagtcct gatcatgctg ctgcactcaa tcttggacaa 2580cagagcaaga ccctgtctca aaaaaaaaaa aaaaaaatat atatatatat atatattatt 2640tttatgaggt gaagtgcatc aaacttggga aagatttgag gaggctggga acctcctgga 2700aaaccactcc ttgaagaaag atatgagaga catttagaag tgattcctgc tttcagaagg 2760aggtggattc aaatacatca aaagtccctt cctctgctaa gtgtttatag ttcaatgaat 2820aatttcaata tttgtatgtg ttcttgtcat tttatttttt tctgaaaaac ttccaaaaat 2880ttgaaaataa aattacagcc ttttcttctt ataaaaaaaa aaaaaaaa 2928801093DNAHomo sapiens 80attgcattcc tgggcattgc taactagtga agtataccag atggaaatgt cttcgaagct 60gtccctttaa aactcgagca agctaccagg caaactccgc ctccagggag gttccttatt 120aaataggagc caactggctg ggtcggggct caatacccca agcaatacct gcaactgagg 180attcttcccg gggagaccgc agcccatcgg catggctcaa gagtttgtga actgcaaaat 240ccagcctggg aaggtggttg tgttcatcaa gcccacctgc ccgtactgca ggagggccca 300agagatcctc agtcaattgc ccatcaaaca agggcttctg gaatttgtcg atatcacagc 360caccaaccac actaacgaga ttcaagatta tttgcaacag ctcacgggag caagaacggt 420gcctcgagtc tttattggta aagattgtat aggcggatgc agtgatctag tctctttgca 480acagagtggg gaactgctga cgcggctaaa gcagattgga gctctgcagt aaccacagat 540ctcataggaa atgttcaaca attctgtgaa aggtcacagg acccaattgg agaaatcata 600tgaaaagcat agttggtctt ggtgtcatat ggatcagagg cacaagtgca gaggctgtgg 660tcatgcggaa cactctgtta tttaagatgg ctatccagat aatcctgaac actgtgtatt 720tattttattt agactaccag caaagattaa agcatgaaat gtaaaacatc tgataaaact 780tacagccccc tacaccaaga gtgtatctgt gaaagagctc ctacactttg aaaacttaag 840aatcccttat catgaagttt gcctgttcta gaattgtaag attgttaatt tccttcaatc 900tctagtgaca acacttaatt tcttttctaa taaaaaaaac ctatagatga ttcagtgatt 960tttgtccaat tcatttgcat gttctcaaga cattaaggaa tgttatgcga aatacactaa 1020cttaaaactg tgtttatatt tggccctgcc attataaata aagacacgtg ctgctgtcaa 1080aaaaaaaaaa aaa 109381827DNAHomo sapiens 81gcagtggagg cggcccaggc ccgccttccg cagggtgtcg ccgctgtgcc gctagcggtg 60ccccgcctgc tgcggtggca ccagccagga ggcggagtgg aagtggccgt ggggcgggta 120tgggactagc tggcgtgtgc gccctgagac gctcagcggg ctatatactc gtcggtgggg 180ccggcggtca gtctgcggca gcggcagcaa gacggtgcag tgaaggagag tgggcgtctg 240gcggggtccg cagtttcagc agagccgctg cagccatggc cccaatcaag gtgggagatg 300ccatcccagc agtggaggtg tttgaagggg agccagggaa caaggtgaac ctggcagagc 360tgttcaaggg caagaagggt gtgctgtttg gagttcctgg ggccttcacc cctggatgtt 420ccaaggttcg gctcctggct gatcccactg gggcctttgg gaaggagaca gacttattac 480tagatgattc gctggtgtcc atctttggga atcgacgtct caagaggttc tccatggtgg 540tacaggatgg catagtgaag gccctgaatg tggaaccaga tggcacaggc ctcacctgca 600gcctggcacc caatatcatc tcacagctct gaggccctgg gccagattac ttcctccacc 660cctccctatc tcacctgccc agccctgtgc tggggccctg caattggaat gttggccaga 720tttctgcaat aaacacttgt ggtttgcggc caaaaaaaaa aaaaaaaaaa aaaaaaaaaa 780aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaa 82782692DNAHomo sapiens 82gcagtggagg cggcccaggc ccgccttccg cagggtgtcg ccgctgtgcc gctagcggtg 60ccccgcctgc tgcggtggca ccagccagga ggcggagtgg aagtggccgt ggggcgggta 120tgggactagc tggcgtgtgc gccctgagac gctcagcggg ctatatactc gtcggtgggg 180ccggcggtca gtctgcggca gcggcagcaa gacggtgcag tgaaggagag tgggcgtctg 240gcggggtccg cagtttcagc agagccgctg cagccatggc cccaatcaag gttcggctcc 300tggctgatcc cactggggcc tttgggaagg agacagactt attactagat gattcgctgg 360tgtccatctt tgggaatcga cgtctcaaga ggttctccat ggtggtacag gatggcatag 420tgaaggccct gaatgtggaa ccagatggca caggcctcac ctgcagcctg gcacccaata 480tcatctcaca gctctgaggc cctgggccag attacttcct ccacccctcc ctatctcacc 540tgcccagccc tgtgctgggg ccctgcaatt ggaatgttgg ccagatttct gcaataaaca 600cttgtggttt gcggccaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 660aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aa 692831537DNAHomo sapiens 83ccctgcgtct ctgcccgccc cgtggcgccc gagtgcactg aagatggcgg ctgctgtagg 60acggttgctc cgagcgtcgg ttgcccgaca tgtgagtgcc attccttggg gcatttctgc 120cactgcagcc ctcaggcctg ctgcatgtgg aagaacgagc ttgacaaatt tattgtgttc 180tggttccagt caagcaccct attttaaggg tacagccgtt gtcaatggag agttcaaaga 240cctaagcctt gatgacttta aggggaaata tttggtgctt ttcttctatc ctttggattt 300cacctttgtg tgtcctacag aaattgttgc ttttagtgac aaagctaacg aatttcacga 360cgtgaactgt gaagttgtcg cagtctcagt ggattcccac tttagccatc ttgcctggat 420aaatacacca aggaagaatg gtggtttggg ccacatgaac atcgcactct tgtcagactt 480aactaagcag atttcccgag actacggtgt gctgttagaa ggttctggtc ttgcactaag 540aggtctcttc ataattgacc ccaatggagt catcaagcat ttgagcgtca acgatctccc 600agtgggccga agcgtggaag aaaccctccg cttggtgaag gcgttccagt atgtagaaac 660acatggagaa gtctgcccag cgaactggac accggattct cctacgatca agccaagtcc 720agctgcttcc aaagagtact ttcagaaggt aaatcagtag atcacccatg tgtatctgca 780ccttctcaac tgagagaaga accacagttg aaacctgctt ttatcatttt caagatggtt 840atttgtagaa ggcaaggaac caattatgct tgtattcata agtattactc taaatgtttt 900gtttttgtaa ttctggctaa gaccttttaa acatggttag ttgctagtac aaggaatcct 960ttattggtaa catcttggtg gctggctagc tagtttctac agaacataat ttgcctctat 1020agaaggctat tcttagatca tgtctcaatg gaaacactct tctttcttag ccttacttga 1080atcttgccta taataaagta gagcaacaca cattgaaagc ttctgatcaa cggtcctgaa 1140attttcatct tgaatgtctt tgtattaaac tgaattttct tttaagctaa caaagatcat 1200aattttcaat gattagccgt gtaactcctg caatgaatgt ttatgtgatt gaagcaaatg 1260tgaatcgtat tattttaaaa agtggcagag tgacttaact gatcatgcat gatccctcat 1320ccctgaaatt gagtttatgt agtcatttta cttattttat tcattagcta actttgtcta 1380tgtatatttc tagatattga ttagtgtaat cgattataaa ggatatttat caaatccagg 1440gattgcattt tgaaattata attattttct ttgctgaagt attcattgta aaacatacaa 1500aataaacata ttttaaaaca tttgcatttt accacca 153784964DNAHomo sapiens 84gagcgctctg gagggcgtgg ccgtgggaaa ggaggcgcgg aaagccgacg cgcgtccatt 60ggtcggctgg acgaggggag gagccgctgg ctcccagccc cgccgcgatg agcctcggcc 120gcctttgccg cctactgaag ccggcgctgc tctgtggggc tctggccgcg cctggcctgg 180ccgggaccat gtgcgcgtcc cgggacgact ggcgctgtgc gcgctccatg cacgagtttt 240ccgccaagga catcgacggg cacatggtta acctggacaa gtaccggggc ttcgtgtgca 300tcgtcaccaa cgtggcctcc cagtgaggca agaccgaagt aaactacact cagctcgtcg 360acctgcacgc ccgatacgct gagtgtggtt tgcggatcct ggccttcccg tgtaaccagt 420tcgggaagca ggagccaggg agtaacgaag agatcaaaga gttcgccgcg ggctacaacg 480tcaaattcga tatgttcagc aagatctgcg tgaacgggga cgacgcccac ccgctgtgga 540agtggatgaa gatccaaccc aagggcaagg gcatcctggg aaatgccatc aagtggaact 600tcaccaagtt tggacaccgt ctctccacag ttcctcatcg acaagaacgg ctgcgtggtg 660aagcgctacg gacccatgga ggagcccctg gtgatagaga aggacctgcc ccactatttc 720tagctccaca agtgtgtggc cccgcccgag cccctgccca cgcccttgga gccttccacc 780ggcactcatg acggcctgcc tgcaaacctg ctggtggggc agacccgaaa atccagcgtg 840caccccgccg gaggaaggtc ccatggcctg ctgggcttgg ctcggcgccc ccacccctgg 900ctaccttgtg ggaataaaca gacaaattag cctgctggaa aaaaaaaaaa aaaaaaaaaa 960aaaa 964851031DNAHomo sapiens 85agtcctgact acggcctccg ggccctttgt ccccgctagc ggcgctcggg gtgggggagc 60caggaggggc gggagacggg cgggtatggg ccgcgcgggc gcaggctccc ccgggcgccg 120caggcagcgg tgccagagcc ggggcaggcg gcggccgcga gcccctcggc ggcggaaggc 180cccagcgtgc aggcgcagga gggcgcggcg ccggcggaag aagccctgtc cccgcagctt 240gcgaccggag atccacgaat gtcccaagtc ccaggacccg tgcgcgtccc gggacgactg 300gcgctgtgcg cgctccatgc acgagttttc cgccaaggac atcgacgggc acatggttaa 360cctggacaag taccggggct tcgtgtgcat cgtcaccaac gtggcctccc agtgaggcaa 420gaccgaagta aactacactc agctcgtcga cctgcacgcc cgatacgctg agtgtggttt 480gcggatcctg gccttcccgt gtaaccagtt cgggaagcag gagccaggga gtaacgaaga 540gatcaaagag ttcgccgcgg gctacaacgt caaattcgat atgttcagca agatctgcgt 600gaacggggac gacgcccacc cgctgtggaa gtggatgaag atccaaccca agggcaaggg 660catcctggga aatgccatca agtggaactt caccaagttc ctcatcgaca agaacggctg 720cgtggtgaag cgctacggac ccatggagga gcccctggtg atagagaagg acctgcccca 780ctatttctag ctccacaagt gtgtggcccc gcccgagccc ctgcccacgc ccttggagcc 840ttccaccggc actcatgacg gcctgcctgc aaacctgctg gtggggcaga cccgaaaatc 900cagcgtgcac cccgccggag gaaggtccca tggcctgctg ggcttggctc ggcgccccca 960cccctggcta ccttgtggga ataaacagac aaattagcct gctggaaaaa aaaaaaaaaa 1020aaaaaaaaaa a 1031

Claims

1. A composition comprising at least 12 oligonucleotides chosen among a library of 48 oligonucleotides comprising or consisting of the nucleic acid sequences SEQ ID NO: 25-72.

2. The composition according to claim 1, wherein said composition comprises at least the 12 oligonucleotides that comprise or consist of the nucleic acid sequences SEQ ID NO: 25-36.

3. A kit comprising at least 12 oligonucleotides chosen among a group of 48 oligonucleotides comprising or consisting of the nucleic acid sequences SEQ ID NO: 25-72, said at least 12 oligonucleotides comprising or consisting of the nucleic acid sequences SEQ ID NO: 25-36.

4. A method for diagnosis, and/or the classification, preferably in vitro, of an hematological disorder, in particular myeloid and/or lymphoid hematological disorder, preferably myeloid hematological disorder comprising, comprising measuring an expression level of a gene using the composition according to claim 1.

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