BIOMARKERS OF THE CHANCES OF SURVIVAL OF A PATIENT WITH AML

Abstract

The present invention relates to a method for in vitro or ex vivo diagnosis of the chances of survival of a patient suffering from Acute Myeloid Leukemia (AML) comprising a step of detecting the expression of at least newly identified AML biomarker genes.

Description

Title of the invention: BIOMARKERS OF THE CHANCES OF SURVIVAL OF A PATIENT WITH AML FIELD OF THE INVENTION

[0001] The present invention falls within the context of the diagnosis and treatment of Acute Myeloid Leukemia (AML) and relates to a new method for in vitro or ex vivo diagnosis of the chances of survival of a patient suffering from AML. The invention also relates to a kit and the use of biomarkers for such in vitro or ex vivo diagnosis. STATE OF THE ART

[0002] Acute Myeloid Leukemia (AML - also known as acute myeloblastic leukemia) is a group of malignant hematological cancers affecting the hematopoietic cells of the bone marrow. They are characterized by an overproduction of blast cells (also called "immature blood cells") which are characterized by an inability to differentiate into mature cells and by uncontrolled proliferation. This dysfunction of the bone marrow consequently prevents the normal production of blood cells and results in various clinical syndromes, which can sometimes be very serious and lead to the death of the patient. AML typically occurs in adults over the age of 40, particularly in the elderly (over 60). However, they also affect younger patients and are even listed among pediatric cancers (they are called: childhood AML).

[0003] The diagnosis of AML is based mainly on a blast rate above 20% in the bone marrow or on the presence of certain well-identified genetic abnormalities, such as t(8;21), inv(16) or t(15;17) (Pelcovits and Niroula 2020 PMID 32236160). The choice of therapeutic treatment for patients with AML strongly depends on the molecular and cytogenetic profile of the leukemia (Dinardo and Lachowies 2019 PMID 31350639), as well as the clinical characteristics of the patient (age, comorbidities). Such treatment generally includes induction cytotoxic chemotherapy or therapy based on hypomethylating agents, sometimes complemented by targeted therapy. In addition, bone marrow transplantation (also called “allogous hematopoietic stem cell transplantation”) is also proposed as consolidation treatment, with the aim of avoiding relapse after chemotherapy.An allograft significantly increases the chances of recovery for patients. However, it is an invasive technique that can be difficult for the patient to tolerate and carries a significant risk of graft-versus-recipient disease. Indeed, on average, only 50% of AML patients survive more than a year after the first diagnosis of the disease. This percentage even drops to around 27% of patients surviving, on average, 5 years after this first diagnosis. In addition, the chances of survival decrease drastically the older the patients. Therefore, in order to propose an optimal treatment protocol adapted to each patient, healthcare teams need reliable indicators to estimate the patient's survival prognosis at the time of diagnosis.

[0004] The World Health Organization (WHO) classifies AML into several subtypes, based on their hematological, morphological, cytogenetic (karyotype) characteristics and genetic mutations (Arber et al. 2016 PM1D 27069254). Based on this classification, the European LeukemiaNet (ELN) expert consortium has proposed a prognostic classification system with three risk levels: favorable, intermediate and adverse (Dôhner et al. 2022 PM1D 35797463). Although this classification is commonly used in clinical practice, it is not sufficient to define a therapeutic strategy applicable to all patient cases, for example, to make the decision for allogeneic transplantation for patients in the intermediate risk category.Therefore, the search for new indicators to clarify the survival prognosis of patients with AML is therefore necessary to enable the refinement of existing classifications and thus enable the most appropriate treatment to be proposed for each patient.

[0005] In this sense, and although the diagnoses relating to AML have improved considerably in recent decades, these do not take into consideration transcriptomic data on gene expression. However, these provide new and complementary information to existing classifications and represent, as such, a promising source of indicators in the prognosis of the chances of survival of a patient suffering from cancer.

[0006] In this context, drawing on the work of researchers at the Institute for Advanced Biosciences (IAB) in Grenoble - who demonstrated that during oncogenesis, the cell undergoes severe deregulation at the genome and epigenome levels that impact gene expression profiles - the inventors unexpectedly and surprisingly identified five new biomarkers by focusing on diagnosing the chances of survival of patients with AML. More specifically, the inventors developed a novel method and an associated diagnostic tool to propose new biomarkers capable, alone or in combination, of predicting the chances of survival of a patient with AML. The inventors tested their method on numerous cohorts of patients with AML, validating it for each of them. In addition, the inventors demonstrated that the prediction of the chances of survival obtained via this new method represents an indicator reliable and robust, as such, and can also be used in addition to indicators already known in the prior art in order to refine existing classifications. DESCRIPTION OF THE INVENTION

[0007] The subject of the present invention is therefore a method for in vitro or ex vivo diagnosis of the chances of survival of a patient suffering from Acute Myeloid Leukemia (AML), comprising a step of detecting the expression of at least one gene chosen from: ADSS1, FAM171B, MLF1, SLITRK5 and TNNT1, in a biological sample of said patient.

[0008] Thus, via the detection of an ectopic expression of at least one of the aforementioned genes, in a biological sample, the method of the invention makes it possible to provide a robust indicator, as such, to the care teams regarding the chances of survival of the patient suffering from AML to be treated. In addition, this additional indicator can be combined with existing ones (ELN indicator, indicator linked to the age of the patient), thus reinforcing the robustness of the established prediction. This indicator also has the advantage of being able to be obtained quickly and simply, via technologies already deployed in a large number of medical centers, for example by an RT-qPCR test. Consequently, obtaining this additional indicator via the method according to the invention is inexpensive and can be standardized in the short term, in the laboratories of said medical centers.

[0009] The present invention makes a real contribution to the panel of Acute Myeloid Leukemia (AML) diagnoses available to healthcare teams in order to offer the most appropriate therapeutic treatment for each patient suffering from AML.

[0010] In the context of the invention, the expressions "in vitro diagnosis" and "ex vivo diagnosis" are understood to refer to tests carried out outside the patient, after taking the biological sample from said patient. The tests carried out on biological samples taken from the patient(s) and kept in the laboratory, generally under sterile conditions, without modification over a short period (generally less than 24 hours), fall within the scope of an ex vivo diagnosis according to the invention. The tests carried out on such samples and over a longer period of time generally fall within the scope of an in vitro diagnosis according to the invention, since they require cell culture steps aimed at keeping the cells of said biological samples taken alive.

[0011] In the context of the invention, the expression "biological sample" refers to a tissue, a fluid, as well as components of said tissue and fluid (for example cells) likely to contain cancerous blast cells. According to the field of application of the invention and by way of non-exhaustive examples, the biological sample may be a bone marrow sample or a blood sample, in particular a peripheral blood sample.

[0012] In the context of the invention, the expressions “chance(s) of survival”, “survival prognosis”, “vital prognosis”, “probability(s) of survival” or “survival prediction(s)” are synonyms and all designate the estimation of the survival of a patient suffering from AML, with regard to indicators specific to this patient.

[0013] In the context of the invention, the expression "detection of the expression of at least one gene" means the quantification of at least one gene expression product. The expression product of a gene, within the meaning of the present invention, is any biological molecule resulting from the expression of said gene. For example, the gene expression product may be an RNA transcript or a protein obtained after translation of a transcript by ribosomes. By "transcript", we mean RNA, and in particular messenger RNA (mRNA), resulting from the transcription of the gene. More precisely, transcripts are RNA produced by the transcription of a gene followed by post-transcriptional modifications of the pre-RNA forms. In the context of the present invention, it is possible to quantify the level of expression of one or more RNA transcripts of the same gene and / or the level of expression of one or more isoforms of a protein encoded by the same gene.In certain cases, the quantified expression of said detected gene can be compared to a reference value. Below this value, the gene is considered not to be expressed, within the meaning of the invention. It may for example be a random and / or isolated expression of said gene, representing background noise, which should not be taken into consideration within the scope of the invention. Conversely, above the reference value, the expression of the gene is considered to be detected. This is then an ectopic expression, also called out-of-context expression, since the five genes identified by the inventors are not expressed in the blast cells of a healthy individual. The detection of gene expression is carried out by any means known to those skilled in the art.In the case of an mRNA transcript, detection may be carried out by a direct method, by any method known to those skilled in the art for determining the presence of said transcript in the sample, or by indirect detection of the transcript after transformation of the latter into DNA, or after amplification of said transcript or after amplification of the DNA obtained after transformation of said transcript into DNA. Many methods exist for the detection of nucleic acids (see for example Kricka et al., Clinical Chemistry, 1999, No. 45(4), p.453-458; Relier GH et al., DNA Probes, 2nd Ed., Stockton Press, 1993, sections 5 and 6, p.173-249).Gene expression can be measured, in particular, by Reverse Transcription-Polymerase Chain Reaction or RT-PCR, preferably by quantitative RT-PCR or RT-qPCR (for example using FilmArray® technology or Fluidigm's BiomarkTM platform), by sequencing (preferably by high-throughput sequencing) or by hybridization techniques (for example with hybridization microarrays or by techniques such as NanoString® nCounter®).

[0014] In the context of the invention, the expression “reference value” corresponds to the value of the level of expression of a gene considered in one or more healthy individuals, i.e. not suffering from AML and / or not presenting any specific medical condition. This reference value can be determined directly from the analysis of a biological sample from a healthy individual or be the result of several values ​​from several biological samples from the same healthy individual and / or from several healthy individuals.

[0015] In the context of the invention, the ADSS1 gene refers to the gene known as "adenylosuccinate synthase 1" (Ensembl identifier: ENSG00000185100) and located on chromosome 14 (chromosomal location of the gene according to GRCh38 / hg38: chrl4: 104,724,174-104,747,325). It has the nucleotide sequence SEQ ID NO: 1 or a sequence having at least 65% or even 70%, 75%, 80%, 85%, 90%, 95% or even 99% identity with the sequence SEQ ID NO: 1.

[0016] In the context of the invention, the FAM171B gene refers to the gene known as "family with sequence similarity 171 member B" (Ensembl identifier: ENSG00000144369) and located on chromosome 2 (chromosomal location of the gene according to GRCh38 / hg38: chr2: 186,694,060-186,765,959). It has the nucleotide sequence SEQ ID NO: 2 or a sequence having at least 65% or even 70%, 75%, 80%, 85%, 90%, 95% or even 99% identity with the sequence SEQ ID NO: 2.

[0017] In the context of the invention, the MLF1 gene refers to the gene known as "Myeloid Leukemia Factor 1" (Ensembl identifier: ENSG00000178053) and located on chromosome 3 (chromosomal location of the gene according to GRCh38 / hg38: chr3: 158,571,163-158,607,252). It has the nucleotide sequence SEQ ID NO: 3 or a sequence having at least 65% or even 70%, 75%, 80%, 85%, 90%, 95% or even 99% identity with the sequence SEQ ID NO: 3.

[0018] In the context of the invention, the SLITRK5 gene refers to the gene known as "SLIT and NTRK like family member 5" (Ensembl identifier: ENSG00000165300) and located on chromosome 13 (chromosomal location of the gene according to GRCh38 / hg38: chrl3: 87,671,371-87,696,272). It has the nucleotide sequence SEQ ID NO: 4 or a sequence having at least 65% or even 70%, 75%, 80%, 85%, 90%, 95% or even 99% identity with the sequence SEQ ID NO: 4.

[0019] In the context of the invention, the TNNT1 gene refers to the gene known as "troponin Tl, slow skeletal type" (Ensembl identifier: ENSG00000105048) and located on chromosome 19 (chromosomal location of the gene according to GRCh38 / hg38: chrl9: 55,132,698-55,149,206). It has the nucleotide sequence SEQ ID NO: 5 or a sequence having at least 65% or even 70%, 75%, 80%, 85%, 90%, 95% or even 99% identity with the sequence SEQ ID NO: 5.

[0020] Preferably, the subject of the present invention is a method as described above and having the following technical characteristics, taken alone or in combination: - the step of detecting the expression of at least one gene chosen from: ADSS1, FAM171B, MLF1, SLITRK5 and TNNT1 consists of measuring the expression of said gene and comparing said measurement with a reference value of expression of said gene for a healthy individual,

[0021] - detection of gene expression is performed via RT-qPCR technology or via high-throughput RNA-seq sequencing,

[0022] - the detection step comprises detecting the expression of at least two of said genes, preferably at least three of said genes, preferentially at least four of said genes, even more preferentially five genes,

[0023] - the detection step comprises detecting the expression of two selected genes among: FAM171B, MLF1 and SLITRK5,

[0024] - the detection step comprises the detection of the expression of the following three genes: FAM171B, MLF1 and SLITRK5,

[0025] - the detection step comprises detecting the expression of the four genes following: FAM171B, MLF1, SLITRK5 and TNNT1,

[0026] - the detection step comprises the detection of the expression of the five genes,

[0027] - the patient's biological sample is a bone marrow sample of said patient, and

[0028] - which the patient is aged 50 to 80 years, preferably 60 to 70 years.

[0029] The invention also relates to a kit for the in vitro or ex vivo diagnosis of the chances of survival of a patient suffering from Acute Myeloid Leukemia (AML) comprising means for detecting the expression of at least one gene chosen from: ADSS1, FAM171B, MLF1, SLITRK5 and TNNT1, in a biological sample of said patient.

[0030] Preferably, the detection means of the kit according to the invention allow the detection of the expression of at least one gene chosen from: ADSS1, FAM171B, MLF1, SLITRK5 and TNNT1 in a bone marrow sample from said patient.

[0031] The invention also relates to a use of at least one gene from: ADSS1, FAM171B, MLF1, SLITRK5 and TNNT1, as a biomarker for the in vitro or ex vivo diagnosis of the chances of survival of a patient suffering from Acute Myeloid Leukemia (AML), in a biological sample from said patient.

[0032] Preferably, the present invention relates to a use as described above and having the following technical characteristics, taken alone or in combination: - the use of at least two of said genes, preferably at least three of said genes genes, preferably at least four of said genes, even more preferably five genes, as a biomarker for the in vitro or ex vivo diagnosis of the chances of survival of a patient suffering from Acute Myeloid Leukemia (AML), in a biological sample of said patient, and

[0033] - the patient's biological sample is a bone marrow sample of said patient. Brief description of the drawings

[0034] [Fig.l]: represents the average survival over time of patients with AML and belonging to different cohorts, according to the detection of the expression of the MLF1 gene;

[0035] [Fig.2]: represents the average survival over time of patients with AML and belonging to different cohorts, based on the detection of MLF1 and FAM171B gene expression;

[0036] [Fig.3]: represents the average survival over time of patients with AML and belonging to different cohorts, based on the detection of MLF1 and SLITRK5 gene expression;

[0037] [Fig.4]: represents the average survival over time of patients with AML and belonging to different cohorts, based on the detection of MLF1, FAM171B and SLITRK5 gene expression;

[0038] [Fig.5]: represents the average survival over time of patients with AML and belonging to different cohorts, based on the detection of MLF1, FAM171B, SLITRK5 and TNNT1 gene expression; and

[0039] [Fig.6]: represents the average survival over time of patients with AML and belonging to different cohorts, based on the detection of MLF1, FAM171B, SLITRK5, TNNT1 and ADSS1 gene expression.

[0040] The present invention is illustrated in a non-limiting manner by the following examples. EXAMPLES OF ACHIEVEMENT

[0041] Example 1: identification and validation of the five biomarkers

[0042] The inventors established a list of tissue-specific genes that are not expressed or are weakly expressed in the bone marrow of healthy patients. More than 3000 genes were retained at this stage.

[0043] Among the genes selected, the inventors then carried out a step of selecting genes for which ectopic activation in leukemias was observed in more than 10% of cases. 314 genes were selected at this step.

[0044] In the next step, the inventors identified candidate genes capable of individually predicting the vital prognosis of patients with AML. To do this, they established an activation threshold for each gene among the 314, in a training dataset (corresponding to a fictitious cohort of AML patients) and evaluated their association with survival, considering their expression level to define their activation status (ON: expressed, or OFF: extinguished). The logrank statistical test (a recognized and commonly used test for survival analysis) and the Cox proportional hazard model (a model also recognized in this field and described in the publication "Analysis of survival data. Cox & Oakes. 1984. ISBN 978-0-412-24490-2") were used in the survival analysis that allowed the identification of these candidate genes.

[0045] At the end of this step, the five candidate genes ADSS1, FAM171B, MLF1, SLITRK5 and TNNT1 were retained with regard to values ​​predicting the survival of AML patients, such as the p-value, the risk ratio and the robustness of their ON / OFF activation status. These genes were then validated individually in two independent AML data sets. Example 2: Results

[0046] The five biomarkers retained after the validation phase were used, alone or in various combinations, in multiple cohorts of patients suffering from AML, in order to confirm the robustness of the diagnostic method according to the invention and the associated prognostic tool. The latter make it possible to classify patients according to their number of biomarker genes presenting ectopic expression (therefore having an ON activation status). The greater the number of biomarker genes, among the five biomarker genes ultimately retained, presenting the activated status (ON status), the less favorable the patient's vital prognosis is, as demonstrated by the following results.

[0047] For each dataset (cohort) tested, the patients were separated into two groups according to their activation status of the chosen gene(s). The difference in survival probabilities between the two established groups was calculated by the logrank test and was found to be statistically significant in all the datasets considered (logrank p-value < 0.05). The impact of the number of activated genes, among the five biomarker genes, on survival was also calculated by the Cox model. The inventors proved that the number of activated genes is significantly correlated with the survival probability of the patients in all the datasets considered (Cox p-value < 0.05), thus demonstrating the robust prediction of the survival chances of patients with acute myeloid leukemia via the method according to the invention and the associated tool.

[0048] The five identified biomarker genes were tested according to different distributions and on the following datasets (cohorts of patients with AML): - public datasets GSE10691, GSE146173, GSE37642, BEATAML and TCGA-LAML (RNA-seq transcriptomic data and DNA microarrays), - public TARGET-AML dataset in which patients are minors and - dataset created by the inventors TRANSCRI-LAM. This cohort is composed of 100 LAM patients who had a diagnosis of LAM, followed at the Grenoble Alpes University Hospital (CHUGA) and who gave their consent for the AC-2014-2094 CRB09 clinical hematology collection. On this cohort, the expression levels of the biomarkers were quantified via high-throughput RNA-seq sequencing (TRANSCRI-LAM-RNAseq variation of this cohort in Figures 1 to 6) or via a standardized RT-qPCR test (TRANSCRI-LAM- RT-qPCR variation of this cohort in Figures 1 to 6). In clinical practice, an RT-qPCR test has several advantages over high-throughput RNA-seq sequencing. It is significantly less expensive, standardizable and widely available in hospital laboratories.The inventors then verified that their method and the associated tool could be advantageously standardized on this technology. Thus, the expression levels of the five biomarker genes were measured via the RT-qPCR technique in all patient samples of the TRANSCRI-LAM cohort. The results were normalized with respect to four housekeeping genes ABL, ACTB, U6 and GAPDH and expressed relative to the expression of the gene in its reference tissue (muscle for the ADSS1 and TNNT1 genes, brain for the FAM171B and SLITRK5 genes and testis for the MLF1 gene). The activation threshold of each gene (definition of the ON / OFF detection status) was established with respect to a reference value obtained using 6 samples of healthy bone marrow and equal to the average of the expressions of said gene in the 6 samples plus two standard deviations. The results show that the method according to the invention (and therefore the associated tool) is well adapted to RT-qPCR technology.A standardized and reproducible RT-qPCR test can therefore be offered in the context of hospital practice.

[0049] 2.1: Use of the MLF1 gene as a biomarker ([Fig. 11)

[0050] In this test, carried out on all the cohorts described previously, the patients of each cohort were differentiated according to:

[0051] - detection of the expression of the MLF1 gene (groups 1 in [Fig.l]) and

[0052] - the absence of detection of the expression of this gene (groups 0 in [Fig.l]),

[0053] in their respective bone marrow samples (this gene being known to be expressed only ectopically in bone marrow cells).

[0054] It is noted for each of the cohorts that patients in group 0 for whom no ectopic expression is detected have better chances of survival than patients in group 1 for whom gene expression is detected.

[0055] 2.2: Use of a combination of the MLF1 and FAM171B genes as bio- markers ([Fig.2])

[0056] In this test, carried out on all the cohorts described previously, the patients of each cohort were differentiated according to:

[0057] - detection of the expression of at least one of the MLF1 or FAM171B genes (groups 1-2 in [Fig.2]) and

[0058] - the absence of detection of the expression of these genes (groups 0 in [Fig.2]),

[0059] in their respective bone marrow samples (these genes being known to be expressed only ectopically in bone marrow cells).

[0060] It is noted for each of the cohorts that the patients in group 0 for whom no ectopic expression is detected have better chances of survival than the patients in group 1-2 for whom the expression of at least one of the two genes is detected.

[0061] 2.3: Use of a combination of the MLF1 and SLITRK5 genes as bio- markers ([Fig.31

[0062] In this test, carried out on all the cohorts described previously, the patients of each cohort were differentiated according to:

[0063] - detection of the expression of at least one of the two MLF1 genes or SLITRK5 (groups 1-2 in [Fig.3]) and

[0064] - the absence of detection of the expression of these genes (groups 0 in [Fig.3]),

[0065] in their respective bone marrow samples (these genes being known to be expressed only ectopically in bone marrow cells).

[0066] It is noted for each of the cohorts that the patients in group 0 for whom no ectopic expression is detected have better chances of survival than the patients in group 1-2 for whom the expression of at least one of the two genes is detected.

[0067] 2.4: Use of a combination of the genes MLF1, FAM171B and SLITRK5 as as biomarkers ([Fig.4D

[0068] In this test, carried out on all the cohorts described previously, the patients of each cohort were differentiated according to:

[0069] - detection of the expression of at least two of the following genes: MLF1, FAM171B or SLITRK5 (groups 2-3 in [Fig.4]) and

[0070] - the absence of detection of the expression of these genes or the detection of the expression of only one of these three genes (groups 0-1 in [Fig.4]),

[0071] in their respective bone marrow samples (these genes being known to be expressed only ectopically in bone marrow cells).

[0072] It is noted for each of the cohorts that the patients in group 0-1 for whom at most the expression of only one of the three genes is detected have better chances of survival than the patients in group 2-3 for whom the expression of at least two of the three genes is detected.

[0073] 2.5: Use of a combination of the genes MLF1. FAM171B. SLITRK5 and TNNT1 as biomarkers ( [Fig.5]}

[0074] In this test, carried out on all the cohorts described previously, the patients of each cohort were differentiated according to:

[0075] - detection of the expression of at least two of the following genes: MLF1, FAM171B, SLITRK5 and TNNT1 (groups 2-4 in [Fig.5]) and

[0076] - the absence of detection of the expression of these genes or the detection of the expression of only one of these four genes (groups 0-1 in [Fig.5]),

[0077] in their respective bone marrow samples (these genes being known to be expressed only ectopically in bone marrow cells).

[0078] It is noted for each of the cohorts that the patients in group 0-1 for whom at most the expression of only one of the four genes is detected have better chances of survival than the patients in group 2-4 for whom the expression of at least two of the four genes is detected.

[0079] 2.6: Use of a combination of the genes MLF1. FAM171B. SLITRK5. TNNT1 and ADSS1 as biomarkers ([Fig.6D

[0080] In this test, carried out on all the cohorts described previously, the patients of each cohort were differentiated according to:

[0081] - detection of the expression of at least two of the following genes MLF1, FAM171B, SLITRK5, TNNT1 and ADSS1 (groups 2-5 in [Fig.6]) and

[0082] - the absence of detection of the expression of these genes or the detection of the expression of only one of these five genes (groups 0-1 in [Fig.6]),

[0083] in their respective bone marrow samples (these genes being known to be expressed only ectopically in bone marrow cells).

[0084] It is noted for each of the cohorts that the patients in group 0-1 for whom at most the expression of only one of the five genes is detected have better chances of survival than the patients in group 2-5 for whom the expression of at least two of the five genes is detected. 2.7: Conclusions

[0085] It is clear from these experimental data that a method, a kit and a use according to the invention make it possible to provide an indicator as to the chances of survival of patients suffering from AML which is 1 / additional and complementary to those already available to medical teams, 2 / robust and reliable as such and 3 / able to be obtained quickly and according to technologies already deployed in a large number of medical centers, for example by an RT-qPCR test. It is also clear from these experimental data, in particular those relating to the TARGET-AML cohort, that the method, the kit and the use according to the invention are applicable to AML in children, since they provide such an indicator for minor patients.

[0086] Furthermore, the data presented also demonstrate that the robustness of this indicator is enhanced the greater the number of the five biomarker genes (whose expression is detected) used.

Claims

Claims

1. Method for in vitro or ex vivo diagnosis of the chances of survival of a patient suffering from Acute Myeloid Leukemia (AML), comprising a step of detecting the expression of at least one gene chosen from: ADSS1, FAM171B, MLF1, SLITRK5 and TNNT1, in a biological sample of said patient.

2. Method according to the preceding claim, in which the step of detecting the expression of at least one gene chosen from: ADSS1, FAM171B, MLF1, SLITRK5 and TNNT1 consists of measuring the expression of said gene and comparing said measurement with a reference value of expression of said gene for a healthy individual.

3. A method according to any preceding claim, wherein the detection of gene expression is performed via RT-qPCR technology or via high-throughput RNA-seq sequencing.

4. A method according to any one of the preceding claims, wherein the detecting step comprises detecting the expression of at least two of said genes, preferably at least three of said genes, preferentially at least four of said genes, even more preferentially all five genes.

5. Method according to the preceding claim, wherein the detection step comprises detecting the expression of two genes chosen from: FAM171B, MLF1 and SLITRK5.

6. The method of claim 4, wherein the detecting step comprises detecting the expression of the following three genes: FAM171B, MLF1 and SLITRK5.

7. The method of claim 4, wherein the detecting step comprises detecting the expression of the following four genes: FAM171B, MLF1, SLITRK5 and TNNT1.

8. The method of claim 4, wherein the detecting step comprises detecting the expression of the five genes.

9. A method according to any preceding claim, wherein the biological sample of the patient is a bone marrow sample of said patient.

10. Kit for the in vitro or ex vivo diagnosis of the chances of survival of a patient suffering from Acute Myeloid Leukemia (AML) comprising means for detecting the expression of at least one gene chosen from: ADSS1, FAM171B, MLF1, SLITRK5 and TNNT1, in a sample biological of said patient.

11. Kit according to the preceding claim, in which the biological sample of the patient is a sample of bone marrow of said patient.

12. Use of at least one gene from: ADSS1, FAM171B, MLF1, SLITRK5 and TNNT1, as a biomarker for the in vitro or ex vivo diagnosis of the chances of survival of a patient suffering from Acute Myeloid Leukemia (AML), in a biological sample of said patient.

13. Use according to the preceding claim of at least two of said genes, preferably at least three of said genes, preferentially at least four of said genes, even more preferentially five genes, as a biomarker for the in vitro or ex vivo diagnosis of the chances of survival of a patient suffering from Acute Myeloid Leukemia (AML), in a biological sample of said patient.

14. Use according to claim 12 or 13, wherein the biological sample of the patient is a bone marrow sample of said patient.

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