CN107974506A

CN107974506A - A kind of method and device for the instruction of acute myeloid leukemia medicine

Info

Publication number: CN107974506A
Application number: CN201810045606.5A
Authority: CN
Inventors: 郭安源; 林生彦; 陈智超; 苗亚茹
Original assignee: Ezhou Institute of Industrial Technology Huazhong University of Science and Technology
Current assignee: Ezhou Institute of Industrial Technology Huazhong University of Science and Technology
Priority date: 2018-01-17
Filing date: 2018-01-17
Publication date: 2018-05-01
Also published as: CN108277278A; CN108130372A

Abstract

An embodiment of the present invention provides a kind of method and device for the instruction of acute myeloid leukemia medicine, method includes：The goal-based assessment gene of normal karyotype acute myeloid leukemia CN AML patients is obtained, the goal-based assessment gene is STAB1 genes；The gene expression amount that gene is assessed according to CN AML patients targets is layered the CN AML patients, and the CN AML patients are divided into prognosis bona's group and prognosis mala group；Corresponding drug information is indicated to the CN AML patients in prognosis mala group；In this way, because goal-based assessment gene only has one, therefore prognosis evaluation and disease layering can be simply carried out to CN AML patients；In addition the goal-based assessment gene is STAB1 genes, because STAB1 genes are membrane protein gene, therefore can be quickly detected using existing detection device, improve detection efficiency.

Description

Method and device for acute myeloid leukemia drug indication

Technical Field

The invention belongs to the technical field of molecular biology, and particularly relates to a method and a device for drug indication of acute myeloid leukemia.

Background

Acute Myelogenous Leukemia (AML) is a malignant clonal disease characterized by abnormal proliferation of immature cells at different stages and reduction of normal hematopoietic tissues due to the obstruction of differentiation of hematopoietic stem cells, and seriously affects the health of human beings.

The AML prognosis can be divided into low-risk, intermediate-risk and high-risk groups based on cytogenetic stratification, wherein the karyotype normal (CN-AML) patients classified into the intermediate-risk group account for about 50% of all AML patients, and such patients have significant heterogeneity and significant difference in prognosis.

Changes in molecular genetics, including genetic mutations and changes in gene expression associated with the prognosis of CN-AML, can now be detected in CN-AML patients, and then used in combination with molecular genetics for a variety of clinical indications, such as: the prognostic evaluation of CN-AML patients can be realized by carrying out combined scoring on gene mutation, patient age, cytogenetic level, gene expression and the like. However, the process of the assessment method is very complicated, disease stratification for prognosis assessment cannot be effectively performed, and corresponding drug information cannot be indicated for patients of corresponding levels, so that a clinical treatment scheme cannot be formulated as soon as possible according to the corresponding drug indication information, and treatment time is delayed.

Disclosure of Invention

Aiming at the problems in the prior art, the embodiment of the invention provides a method and a device for indicating normal karyotype acute myelogenous leukemia drugs, which are used for solving the technical problems that in the prior art, prognosis evaluation and disease stratification cannot be effectively and simply carried out on CN-AML patients, corresponding drug information cannot be indicated for patients of corresponding levels, so that a treatment scheme cannot be rapidly established according to corresponding drug indication in clinic, and treatment time is delayed.

The invention provides a method for drug indication of acute myeloid leukemia, which comprises the following steps:

obtaining a target evaluation gene of a normal karyotype acute myelogenous leukemia CN-AML patient, wherein the target evaluation gene is a STAB1 gene;

the CN-AML patients are stratified according to the gene expression quantity of the target evaluation gene of the CN-AML patients, and the CN-AML patients are divided into a good prognosis group and a bad prognosis group;

corresponding drug information was indicated for CN-AML patients in the poor prognosis group.

In the above embodiment, the stratifying the prognosis survival time of the CN-AML patients according to the gene expression level of the target assessment gene of the CN-AML patients comprises:

obtaining the expression quantity of each gene of the target evaluation gene in a target sample;

determining the median of the expression quantity of each gene;

when the gene expression level of the CN-AML patient target evaluation gene is greater than the median, determining that the CN-AML patient target evaluation gene is a high-expression gene, and classifying the current CN-AML patient into a poor prognosis group;

when the gene expression level of the CN-AML patient target evaluation gene is less than the median, determining that the CN-AML patient target evaluation gene is a low expression gene, and classifying the current CN-AML patient into a good prognosis group.

In the above embodiment, the indicating of the corresponding drug information for the CN-AML patients in the poor prognosis group comprises:

acquiring genes sensitive or resistant to a target drug and semi-inhibitory concentration IC50 values of antagonists of a plurality of drugs in a cancer drug database, wherein the target drug is a clinically empirical drug;

determining the correlation between the expression level of the STAB1 gene and the expression level of the drug-sensitive or drug-resistant gene by utilizing a Pearson function, and if the correlation between the expression level of the STAB1 gene and the expression level of the drug-sensitive or drug-resistant gene is positive, taking the target drug as an indicator drug of a CN-AML patient in a prognosis-poor group;

and determining the correlation between the half-inhibitory concentration IC50 value of the drug antagonist in the cancer drug database and the expression level of the STAB1 gene by using the Pearson function, and if the IC50 value is positively correlated with the expression level of the STAB1 gene, taking the drug as an indicator drug of CN-AML patients in a prognosis-poor group.

cytarabine drug, small molecule inhibitor NVP-BHG712, small molecule inhibitor GSK-J4, small molecule inhibitor BRD-K30748066 and Tozasertib drug information were indicated for CN-AML patients in the poor prognosis group.

In the above scheme, the method includes:

acquiring gene expression information of a target sample from a database, wherein the target sample is a sample of a normal karyotype acute myelogenous leukemia CN-AML patient;

dividing the target sample into a first type sample and a second type sample according to a preset survival time as a classification standard, wherein the first type sample is a sample with the survival time of less than 2 years, and the second type sample is a sample with the survival time of more than 2 years;

screening the genes of the first type sample and the second type sample according to a preset first screening condition to obtain a plurality of differential expression genes;

screening the plurality of differentially expressed genes according to a preset second screening condition to obtain a plurality of prognostically related differentially expressed genes related to the survival time;

obtaining clinical factors influencing the CN-AML prognosis from the database, and carrying out multi-factor proportional risk Cox regression analysis on the multiple prognosis related differential expression genes according to the clinical factors influencing the CN-AML prognosis and empirical prognostic factors influencing the survival of acute myeloid leukemia AML to obtain independent prognostic genes;

and verifying each independent prognostic gene according to a preset verification sample set, and determining the target evaluation gene according to a verification result.

In the above scheme, the obtaining of the gene expression information of the target sample from the database includes:

acquiring gene expression information of a preset number of samples from a cancer gene database TCGA;

and extracting the gene expression information of the target sample from the preset number of samples according to the sample identification of the target sample.

In the above scheme, the screening the genes of the first type of sample and the second type of sample according to a preset first screening condition to obtain a plurality of differentially expressed genes includes:

screening all genes in the first type of sample and all genes in the second type of sample according to the first screening condition to obtain a plurality of differentially expressed genes; wherein the first screening condition is that the false positive rate FDR of the false positive gene is less than 0.05 and the difference fold of the gene fold-change is more than 1.5.

In the foregoing embodiment, the screening the plurality of differentially expressed genes according to a preset second screening condition to obtain a plurality of differentially expressed genes related to prognosis associated with the survival time includes:

obtaining survival data of each differentially expressed gene in the target sample, wherein the survival data comprises: the gene expression quantity of each differential expression gene, the survival time and the survival state of a sample corresponding to each differential expression gene;

based on the survival data of each differential expression gene, performing survival analysis on each differential expression gene by using a curve function in a KM statistical tool to generate a first KM survival curve of each differential expression gene;

obtaining a first significant value of each differentially expressed gene from each first KM survival curve;

screening the first significant value of each differentially expressed gene according to a preset second screening condition to obtain a plurality of prognostically related differentially expressed genes related to the survival time; wherein the second screening condition is that P is less than or equal to 0.05; p is a significant value.

In the above embodiment, the performing a multi-factor proportional risk Cox regression analysis on the plurality of prognosis-related differentially expressed genes according to the clinical factors affecting the CN-AML prognosis and the prognostic factors affecting AML survival to obtain each independent prognosis gene includes:

performing multi-factor proportional risk Cox regression analysis on the plurality of prognosis related differential expression genes according to clinical factors influencing the CN-AML prognosis and prognostic factors influencing the AML survival to obtain a second significant value of each prognosis related differential expression gene;

screening the second significant value of each of the prognosis-related differentially-expressed genes according to a preset second screening condition to obtain each independent prognosis gene; wherein the second screening condition is that P is less than or equal to 0.05; the P is a significant value;

the clinical factors affecting the prognosis of CN-AML include: age, FMS-like tyrosine kinase 3(FLT3) mutation, DNA methyltransferase 3A (DNMT3A) mutation, isochosphate dehydrogenase 1(IDH1) mutation, isochosphate dehydrogenase 2(IDH2) mutation, RUNT-related transcription factor 1(RUNX1) mutation and mitochondrial gene b (mtcyb) mutation, nucleophosmin (NPM1) mutation and williams tumor suppressor 1(WT1) mutation;

the empirical prognostic factors affecting AML survival include: NPM 1mutation, IDH 1mutation, IDH2 mutation and WT1 mutation.

The present invention also provides a device for drug indication of acute myeloid leukemia, the device comprising:

the acquisition unit is used for acquiring a target evaluation gene of a normal karyotype acute myelogenous leukemia CN-AML patient, wherein the target evaluation gene is a STAB1 gene;

the system comprises a layering unit, a screening unit and a screening unit, wherein the layering unit is used for layering CN-AML patients according to the gene expression quantity of a CN-AML patient target evaluation gene and dividing the CN-AML patients into a good prognosis group and a bad prognosis group;

an indicating unit for indicating the corresponding drug information to the CN-AML patients in the poor prognosis group.

The embodiment of the invention provides a method and a device for drug indication of acute myeloid leukemia, wherein the method comprises the following steps: obtaining a target evaluation gene of a normal karyotype acute myelogenous leukemia CN-AML patient, wherein the target evaluation gene is a STAB1 gene; the CN-AML patients are stratified according to the gene expression quantity of the target evaluation gene of the CN-AML patients, and the CN-AML patients are divided into a good prognosis group and a bad prognosis group; indicating corresponding drug information for CN-AML patients in the poor prognosis group; thus, dividing CN-AML patients into a good prognosis group and a bad prognosis group according to the target evaluation genes of the CN-AML patients, and determining the disease stratification of the patients; then according to the corresponding medicine information indicated for CN-AML patients with poor prognosis, a treatment scheme of the poor prognosis group is established in an assisted and rapid clinical manner, so that the treatment opportunity is prevented from being delayed; here, since there is only one target evaluation gene, it is possible to simply perform prognosis evaluation and disease stratification for CN-AML patients; in addition, the target evaluation gene is the STAB1 gene, and the STAB1 gene is a membrane protein gene, so that the rapid detection can be realized by using the existing detection equipment, and the detection efficiency is improved.

Drawings

FIG. 1 is a schematic flow chart of a method for drug indication of acute myelogenous leukemia according to an embodiment of the present invention;

FIG. 2 is a schematic structural diagram of a device for drug indication of acute myelogenous leukemia according to a second embodiment of the present invention;

FIG. 3 is a schematic diagram of the KM survival curve of STATB 1 gene in TCGA CN-AML prediction sample according to the third embodiment of the present invention;

FIG. 4 is a schematic diagram of the KM survival curve of STAB1 gene in GSE12417A prediction sample according to the third embodiment of the present invention;

FIG. 5 is a schematic diagram of the KM survival curve of STAB1 gene in GSE71014 prediction sample provided in example III of the present invention;

FIG. 6 is a schematic diagram of the KM survival curve of STAB1 gene in GSE6891 prediction sample according to the third embodiment of the present invention.

Detailed Description

In order to solve the technical problems that in the prior art, disease stratification and drug indication for prognosis of CN-AML patients cannot be effectively and simply carried out, so that a clinical treatment scheme cannot be formulated as soon as possible according to corresponding drug indication, and the treatment opportunity is delayed, the embodiment of the invention provides a method and a device for drug indication of acute myeloid leukemia, wherein the method comprises the following steps: obtaining a target evaluation gene of a CN-AML patient, wherein the target evaluation gene is a STAB1 gene; the CN-AML patients are stratified according to the gene expression quantity of the target evaluation gene of the CN-AML patients, and the CN-AML patients are divided into a good prognosis group and a bad prognosis group; corresponding drug information was indicated for CN-AML patients in the poor prognosis group.

The technical solution of the present invention is further described in detail by the accompanying drawings and the specific embodiments.

Example one

This example provides a method for drug indication of acute myeloid leukemia, as shown in fig. 1, comprising:

s111, obtaining a target evaluation gene of a normal karyotype acute myelogenous leukemia CN-AML patient, wherein the target evaluation gene is a STAB1 gene;

in this step, before obtaining the target evaluation gene of the normal karyotype acute myelogenous leukemia CN-AML patient, it is necessary to determine which gene the target evaluation gene is, and the specific determination method is as follows:

in this step, gene expression information of a predetermined number of samples is downloaded from a Cancer gene database (TCGA), The predetermined samples including: gene expression information of normal nuclear acute myelogenous leukemia CN-AML patients and abnormal nuclear acute myelogenous leukemia patients, the gene expression information comprises: the amount of gene expression.

Here, since the sample identifications of the CN-AML patient and the abnormal karyotype acute myelogenous leukemia patient are different in the data, the gene expression information of the target sample can be extracted from the predetermined number of samples according to the sample identification of the target sample. The target sample is gene expression information of CN-AML patients.

After a target sample is obtained, dividing the target sample into a first type sample and a second type sample according to a preset survival time as a classification standard; the preset survival time is an experience index for completely relieving CN-AML clinically, and is specifically 2 years. In this embodiment, the first type of samples are samples whose survival time is less than 2 years, and the second type of samples are samples whose survival time is greater than 2 years.

After a first type sample and a second type sample are obtained, screening all genes in the first type sample and all genes in the second type sample by using a Deseq function of a statistical modeling tool R package according to the first screening condition to obtain a plurality of differential expression genes; wherein the first screening condition is that false positive rate (FDR) of false positive gene is less than 0.05 and fold-change of gene is > 1.5.

And after obtaining the differential expression genes, screening the differential expression genes according to a preset second screening condition to obtain a plurality of prognosis-related differential expression genes related to the survival time.

Specifically, survival data of each differentially expressed gene in the target sample is obtained, wherein the survival data comprises: the gene expression quantity of each differential expression gene, the survival time and the survival state of a sample corresponding to each differential expression gene; the living state is a living or dead state, and the living state may correspond to a living state of 1 and the dead state may correspond to a dead state of 0.

Based on the survival data of each differential expression gene, performing survival analysis on each differential expression gene by using a curve function survivval in the R packet to generate a first KM survival curve of each differential expression gene;

For example, taking gene a as an example, gene a has a gene expression level in each target sample, determining the median (median) of the gene expression levels in the target samples, and using Log Rank test function in KM to distinguish between samples with expression levels greater than the median and samples with expression levels less than the median, thereby generating a table; the samples with the expression quantity larger than the median are high expression quantity samples, and the samples with the expression quantity smaller than the median are low expression quantity samples.

And then generating a first KM survival curve of the gene A by using an R-package survivval function according to the survival data of the gene A, reading a significant value P of the gene A from the first KM survival curve of the gene A, and determining the gene A as a differential expression gene related to the survival time when the P value of the gene A is less than or equal to 0.05.

And after determining the plurality of prognosis related differential expression genes related to the survival time, carrying out multi-factor proportional risk Cox regression analysis on the plurality of prognosis related differential expression genes according to clinical factors influencing the CN-AML prognosis and prognostic factors influencing the acute myeloid leukemia AML survival to obtain each prognostic gene.

Specifically, clinical factors affecting the prognosis of CN-AML need to be screened first.

There is also a need to obtain clinical factors from the database that affect the prognosis of the CN-AML. The method comprises the following specific steps: clinical information of the target sample is obtained from the database, and clinical factors with significant statistical significance are screened out by utilizing the survivval function of the R packet and combining the clinical information of the target sample. When the p of the clinical factor is less than or equal to 0.1, the clinical factor is considered to have statistical significance, namely the clinical factor can be used as the clinical factor influencing the CN-AML prognosis. Clinical factors affecting the prognosis of CN-AML in this example include: age, FMS-like tyrosine kinase 3(FLT3) mutation, DNA methyltransferase 3A (DNMT3A) mutation, isophosphate dehydrogenase 1(IDH1) mutation, isophosphate dehydrogenase 2(IDH2) mutation, RUNT-related transcription factor 1(RUNX1) mutation, and mitochondrial gene b (mtcyb) mutation, nucleophosmin (NPM1) mutation, and williams tumor suppressor 1(WT1) mutation.

And then combining literature and clinically identified empirical prognostic factors influencing AML survival, wherein the prognostic factors influencing AML survival comprise: nucleophosmin (NPM1) mutation, isophosphorate dehydrogenase 1(IDH1) mutation, isophosphorate dehydrogenase 2(IDH2) mutation and Williams tumor suppressor 1(WT1) mutation.

And finally, carrying out multi-factor proportional risk Cox regression analysis on the plurality of the prognosis related differential expression genes according to clinical factors influencing the CN-AML prognosis and prognostic factors influencing the survival of acute myeloid leukemia AML, and obtaining a second significant value of each prognosis related differential expression gene.

When the second significance P is less than or equal to 0.05, the gene is an independent prognostic gene independent of Age, FLT3 mutation, DNMT3A mutation, IDH 1mutation, RUNX 1mutation, MT _ CYB mutation, NPM 1mutation, IDH2 mutation and WT1 mutation.

After the independent prognostic genes are obtained, verifying each independent prognostic gene respectively according to a preset verification sample set, and determining a target evaluation gene according to a verification result. The validation sample set may be obtained from the GEO data platform of the NCBI website (https:// www.ncbi.nlm.nih.gov /).

Specifically, obtaining the expression quantity of each independent prognostic gene in the verification sample set respectively, wherein the verification sample set comprises a plurality of verification samples; obtaining survival data of each independent prognostic gene in each validation sample, wherein the survival data comprises: the gene expression quantity of each independent prognosis gene, and the survival time and the survival state of a sample corresponding to each independent prognosis gene; based on the survival data of each independent prognostic gene in each verification sample, performing survival analysis on each prognostic gene by using a curve function in a KM statistical tool to generate a second KM survival curve of each prognostic gene in each verification sample; obtaining a third significant value of each prognostic gene in each validation sample from each second KM survival curve; acquiring the number of third significant values in each verification sample of each independent prognostic gene meeting the second screening condition, wherein the independent prognostic gene corresponding to the third significant value with the largest number meeting the second screening condition is the target evaluation gene; the target evaluation gene is the STAB1 gene.

For example, independent prognostic genes include genes B, C and D; verifying the sample includes: a. b and c; taking gene B as an example, the expression level of each gene in each verification sample of each gene B is obtained, and the survival data in each verification sample is obtained at the same time.

And then according to the survival data of the gene B in each verification sample, respectively generating a third KM survival curve of the gene B by using an R-packet survival function, reading the P values of the gene B in different verification samples from the third KM survival curves of the gene B in different verification samples, and counting the number of the P values meeting a second screening condition, wherein the second screening condition is that the significant value P is less than or equal to 0.05.

Then, the P values of the genes C and D in each verification sample are counted in the same way, and the number of the P values meeting the second screening condition is determined.

Assuming that the number of the genes B whose P values in the respective verification samples satisfy the second screening condition is 3, the number of the genes C whose P values in the respective verification samples satisfy the second screening condition is 2, and the number of the genes D whose P values in the respective verification samples satisfy the second screening condition is 1, the gene B is determined as the target evaluation gene.

Here, the target assessment gene was the STAB1 gene, and the flow meter was used to rapidly obtain the target assessment gene from CN-AML patients, and the STAB1 gene was a membrane protein gene, and thus was rapidly detected using the existing detection equipment.

And S112, layering the CN-AML patients according to the gene expression quantity of the target evaluation gene of the CN-AML patients, and dividing the CN-AML patients into a good prognosis group and a bad prognosis group.

And when the target evaluation gene is determined, acquiring the target evaluation gene of the CN-AML patient, layering the CN-AML patient according to the gene expression quantity of the target evaluation gene of the CN-AML patient, and dividing the CN-AML patient into a good prognosis group and a bad prognosis group.

Here, it is also necessary to first evaluate the accuracy of the predicted survival time of the objective assessment gene. Specifically, each gene expression level of the target evaluation gene in each prediction sample is determined, a median value of the gene expression levels of the target evaluation gene in each prediction sample is determined, a sample with the gene expression level greater than the median value is determined as an evaluation gene high-expression sample, and a gene with the gene expression level less than the median value is determined as an evaluation gene low-expression sample.

Respectively counting a first number of the high-expression samples of the evaluation genes in each prediction sample and a second number of the low-expression samples of the evaluation genes in each prediction sample by taking the preset survival time as a standard; and determining the accuracy of the target evaluation gene in each prediction sample according to the first quantity and the second quantity. Wherein the prediction samples comprise target samples and validation samples.

For example, when determining the accuracy of the gene B in the target sample, the target evaluation gene is the gene B, the sample is firstly divided into an evaluation gene high-expression sample and an evaluation gene low-expression sample according to the gene B, the number of the evaluation gene high-expression samples with the survival time less than 2 years is counted as m, the number of the evaluation gene low-expression samples with the survival time more than 2 years is counted as n, and then the accuracy of the target evaluation gene B in the target sample is calculated as: (m + n)/S; the S is the number of target samples.

When the accuracy is determined, the accuracy of the target evaluation gene is considered to be feasible when the accuracy is 60% or more.

Then it can be considered that the accuracy of the prognostic survival time of CN-AML patients evaluated based on the gene expression level of the target evaluation gene is high.

The CN-AML patients can then be stratified according to their gene expression levels of their target evaluation genes. When the gene expression level of the CN-AML patient target evaluation gene is greater than the median, determining that the CN-AML patient target evaluation gene is a high-expression gene, and classifying the current CN-AML patient into a poor prognosis group;

S113, corresponding drug information is indicated for CN-AML patients in the poor prognosis group.

After the CN-AML patients are divided into a good prognosis group and a bad prognosis group according to the target evaluation gene expression quantity, corresponding medicine information can be indicated for the CN-AML patients in the bad prognosis group so as to assist the clinical designation of an accurate layered treatment scheme.

The method comprises the following specific steps: firstly, acquiring a gene sensitive or resistant to a target drug, wherein the target drug is a clinically empirical drug such as cytarabine; after genes sensitive to or resistant to cytarabine are obtained, the correlation between STAB1 and the expression quantity of the genes sensitive to or resistant to cytarabine is calculated by adopting a Pearson function of an R packet, and the result shows that the high expression of STAB1 is positively correlated with the expression of the genes resistant to cytarabine, so that the high expression sample (poor prognosis group) of STAB1 can reveal that the patients are resistant to cytarabine, namely, the fact that the dosage of cytarabine needs to be increased or other treatment schemes need to be selected in the STAB1 high expression patient group.

Meanwhile, the pearson correlation between the half inhibitory concentration IC50 value of the drug antagonist in the GDSC (drug Sensitivity in cancer) database and the expression level of STAB1 in the CTRP (cancer therapeutics Response Portal) database is respectively calculated, and the drugs with the p value of less than 0.05 (namely the correlation has statistical significance) are obtained, including the drugs with the IC50 value showing positive correlation and negative correlation with the expression level of STAB 1. The positive correlation between the IC50 value and the STAB1 shows that the STAB1 high-expression sample has sensitivity to the drugs, namely that the STAB1 high-expression patients (poor prognosis groups) are sensitive to the drugs and can be clinically considered as candidate drugs/small molecule inhibitors. Here, the STAB1 high-expression patients are sensitive to small molecule inhibitors NVP-BHG712, GSK-J4, BRD-K30748066 and Tozasertib (Tozasertib).

In conclusion, the CN-AML patients with the prognosis survival time of less than 2 years are patients with high expression of STAB1, the patients with the type have drug resistance to cytarabine, and the dosage of cytarabine drugs needs to be increased or other alternative drugs need to be selected in patients with high expression of STAB 1.

Meanwhile, in the high-expression patient of STAB1, the high-expression patient is sensitive to small molecule inhibitors NVP-BHG712, GSK-J4, BRD-K30748066 and Tozasertib (Tozasertib), which indicates that the medicines possibly have certain effects in the high-expression patient of STAB 1. Corresponding drug information is indicated for patients with high expression of STAB1, so that accurate layered treatment schemes can be clinically specified in an auxiliary mode, and treatment opportunity delay is avoided.

Example two

In accordance with a first embodiment, there is provided a device for drug indication of acute myeloid leukemia, as shown in fig. 2, the device comprising: an acquisition unit 21, a layering unit 22, and an instruction unit 23; wherein,

before obtaining the target assessment genes of the normal karyotype acute myelogenous leukemia CN-AML patient, the obtaining unit 21 needs to determine which gene the target assessment genes are, and the specific determination method is as follows:

the acquiring unit 21 is configured to download and acquire gene expression information of a preset number of samples from the cancer gene database TCGA, where the preset samples include: gene expression information of normal nuclear acute myelogenous leukemia CN-AML patients and abnormal nuclear acute myelogenous leukemia patients, the gene expression information comprises: the amount of gene expression.

Because the sample identifications of the CN-AML patient and the abnormal karyotype acute myelogenous leukemia patient are different in the data, the gene expression information of the target sample can be extracted from the preset number of samples according to the sample identification of the target sample. The target sample is the gene expression information of the CN-AML patient.

After the target sample is obtained, the classifying unit 24 is configured to classify the target sample into a first type sample and a second type sample according to a preset survival time as a classification standard; the preset survival time is an experience index for completely relieving CN-AML clinically, and is specifically 2 years. In this embodiment, the first type of samples are samples whose survival time is less than 2 years, and the second type of samples are samples whose survival time is greater than 2 years.

After the first type sample and the second type sample are obtained, the first screening unit 25 is configured to screen all genes in the first type sample and all genes in the second type sample by using a Deseq function of the R package according to a preset first screening condition, so as to obtain a plurality of differentially expressed genes; wherein the first screening condition is that the false positive rate (FDR) of the differentially expressed genes is <0.05 and the fold-difference (fold-change) of the differentially expressed genes is > 1.5.

After obtaining the differentially expressed genes, the second screening unit 26 is configured to screen the plurality of differentially expressed genes according to a preset second screening condition, and obtain a plurality of prognosis-related differentially expressed genes related to the survival time.

For example, taking gene a as an example, gene a has a gene expression level in each target sample, determining a median (median) of the expression levels of gene a in the target samples, and using a Log Rank test function in KM to distinguish between samples with expression levels greater than the median and samples with expression levels less than the median, thereby generating a table; the samples with the expression quantity larger than the median are high expression quantity samples, and the samples with the expression quantity smaller than the median are low expression quantity samples.

After determining the plurality of prognosis-related differential expression genes related to the survival time, the analysis unit 25 is configured to perform multi-factor proportional risk Cox regression analysis on the plurality of prognosis-related differential expression genes according to the clinical factor affecting the CN-AML prognosis and the prognosis factor affecting AML survival, so as to obtain each prognosis gene.

Specifically, the analysis unit 27 first needs to screen clinical factors that affect the prognosis of the CN-AML.

Here, clinical information of the target sample is acquired from the database, and a survivval function of the R packet is used to screen out a statistically significant clinical factor in combination with the clinical information of the target sample. When the p of the clinical factor is less than or equal to 0.1, the clinical factor is considered to have statistical significance, namely the clinical factor can be used as the clinical factor influencing the CN-AML prognosis. Clinical factors affecting the prognosis of CN-AML in this example include: age, FMS-like tyrosine kinase 3(FLT3) mutation, DNA methyltransferase 3A (DNMT3A) mutation, isophosphate dehydrogenase 1(IDH1) mutation, isophosphate dehydrogenase 2(IDH2) mutation, RUNT-related transcription factor 1(RUNX1) mutation, and mitochondrial gene b (mtcyb) mutation, nucleophosmin (NPM1) mutation, and williams tumor suppressor 1(WT1) mutation.

And then combining with literature and clinically recognized prognostic factors influencing AML survival, wherein the prognostic factors influencing AML survival comprise: nucleophosmin (NPM1) mutation, isophosphorate dehydrogenase 1(IDH1) mutation, isophosphorate dehydrogenase 2(IDH2) mutation and Williams tumor suppressor 1(WT1) mutation.

Finally, the analysis unit 27 performs a multi-factor proportional risk Cox regression analysis on the plurality of prognosis related differentially expressed genes according to the clinical factors affecting the CN-AML prognosis and the prognostic factors affecting AML survival, and obtains a second significant value of each of the prognosis related differentially expressed genes.

When the second significance P is less than or equal to 0.05, the gene is an independent prognostic gene independent of Age, FLT3 mutation, DNMT3A mutation, IDH 1mutation, RUNX 1mutation, MTCBB mutation, NPM 1mutation, IDH2 mutation and WT1 mutation.

After the independent prognostic genes are obtained, the verification unit 28 is configured to verify each independent prognostic gene according to a preset verification sample set, and determine a target evaluation gene according to a verification result. The validation sample set may be obtained from the GEO data platform of the NCBI website (https:// www.ncbi.nlm.nih.gov /).

Specifically, the verification unit 28 obtains each gene expression level of each independent prognostic gene in the verification sample set including a plurality of verification samples; obtaining survival data of each independent prognostic gene in each validation sample, wherein the survival data comprises: the gene expression quantity of each independent prognosis gene, and the survival time and the survival state of a sample corresponding to each independent prognosis gene; based on the survival data of each independent prognostic gene in each verification sample, performing survival analysis on each prognostic gene by using a curve function in a KM statistical tool to generate a second KM survival curve of each prognostic gene in each verification sample; obtaining a third significant value of each prognostic gene in each validation sample from each second KM survival curve; acquiring the number of third significant values in each verification sample of each independent prognostic gene meeting the second screening condition, wherein the independent prognostic gene corresponding to the third significant value with the largest number meeting the second screening condition is the target evaluation gene; the target evaluation gene is the STAB1 gene.

For example, independent prognostic genes include genes B, C and D; verifying the sample includes: a. b and c; taking gene B as an example, the verification unit 28 obtains the expression level of each gene in each verification sample of each gene B, and obtains survival data in each verification sample.

After the target evaluation gene is determined, the stratification unit 22 is configured to stratify the CN-AML patients according to the gene expression level of the target evaluation gene of the CN-AML patients, and to classify the CN-AML patients into a good prognosis group and a bad prognosis group.

Here, the evaluation unit 29 also needs to first evaluate the accuracy of the predicted survival time of the objective evaluation gene. Specifically, each gene expression level of the target evaluation gene in each prediction sample is determined, a median value of the gene expression levels of the target evaluation gene in each prediction sample is determined, a sample with the gene expression level greater than the median value is determined as an evaluation gene high-expression sample, and a gene with the gene expression level less than the median value is determined as an evaluation gene low-expression sample.

When the accuracy is determined by the evaluation unit 29, the accuracy of the target evaluation gene is considered to be feasible when the accuracy is 60% or more.

The stratification unit 22 may then stratify the CN-AML patients based on the gene expression levels of their target assessment genes. When the gene expression level of the CN-AML patient target evaluation gene is greater than the median, determining that the CN-AML patient target evaluation gene is a high-expression gene, and classifying the current CN-AML patient into a poor prognosis group;

After the grouping unit 22 determines the CN-AML patients, the indicating unit 23 can indicate the corresponding drug information to the CN-AML patients with poor prognosis so as to assist the clinical specification of an accurate layered treatment scheme and avoid delaying the treatment time.

EXAMPLE III

In practical application, the target evaluation gene of CN-AML can be determined according to the method and the device, and the gene is used for carrying out prognosis stratification on CN-AML and indicating corresponding medicine information, which is concretely as follows:

the method comprises the steps of firstly downloading gene expression information and clinical information of 200 samples from a TCGA database, and then extracting the gene expression information of CN-AML samples from samples with preset quantity according to sample identifications of the CN-AML samples. The number of the CN-AML samples is 79 cases.

Dividing the CN-AML sample into a first type sample and a second type sample according to a preset survival time as a classification standard; the preset survival time is an experience index for completely relieving CN-AML clinically, and is specifically 2 years. In this embodiment, the first type of samples are CN-AML samples with a lifetime less than 2 years, and the second type of samples are CN-AML samples with a lifetime greater than 2 years.

After a first type sample and a second type sample are obtained, screening all genes in the first type sample and all genes in the second type sample by using a Deseq function of an R package to obtain a plurality of differential expression genes; wherein the first screening condition is that the false positive rate (FDR) of the differentially expressed genes is <0.05 and the fold-change of genes is > 1.5. Here, the number of the differentially expressed genes was 353.

After the differentially expressed genes are obtained, performing survival analysis on each differentially expressed gene by using R-package survivval to generate a first KM survival curve of each differentially expressed gene; and obtaining a first significant value of each differentially expressed gene based on the first KM survival curve, and screening the differentially expressed genes which have significant values P less than or equal to 0.05 and are relevant to prognosis related to the survival time, wherein the number of the differentially expressed genes relevant to prognosis is 15.

And then carrying out multi-factor proportional risk Cox regression analysis on the multiple prognosis related differential expression genes according to clinical factors influencing the CN-AML prognosis and prognostic factors influencing the survival of acute myeloid leukemia AML to obtain independent prognostic genes, wherein the number of the independent prognostic genes is 6.

Clinical factors affecting the prognosis of CN-AML in this example include: age, FMS-like tyrosine kinase 3(FLT3) mutation, DNA methyltransferase 3A (DNMT3A) mutation, isophosphate dehydrogenase 1(IDH1) mutation, isophosphate dehydrogenase 2(IDH2) mutation, RUNT-related transcription factor 1(RUNX1) mutation, and mitochondrial gene b (mtcyb) mutation, nucleophosmin (NPM1) mutation, and williams tumor suppressor 1(WT1) mutation.

The prognostic factors affecting AML survival include: nucleophosmin (NPM1) mutation, isophosphorate dehydrogenase 1(IDH1) mutation, isophosphorate dehydrogenase 2(IDH2) mutation and Williams tumor suppressor 1(WT1) mutation.

After the independent prognostic genes are obtained, verifying each independent prognostic gene respectively according to a preset gene chip verification sample set, and determining a target evaluation gene according to a verification result. The validation sample set may be obtained from a GEO platform. The verification sample set comprises four groups, and the number of the samples of each verification sample is as follows: 79. 163, 104 and 187.

Obtaining the significant value P of each independent prognostic gene in each verification sample, and determining the number of P values meeting a second screening condition; the second screening condition is that the significance value P is less than or equal to 0.05. The independent prognostic gene with the largest number of P values which meets the second screening condition is the target evaluation gene. The target evaluation gene in this example was STAB 1.

And then determining the accuracy of the STAB1 by using the prediction sample set, specifically determining each gene expression level of the STAB1 in each prediction sample, determining the median value of the gene expression levels of the STAB1 in each prediction sample, determining the gene with the gene expression level larger than the median value as a high expression evaluation gene, and determining the gene with the gene expression level larger than the median value as a low expression gene.

Respectively counting the first number of the high-expression evaluation genes in each prediction sample and the second number of the low-expression evaluation genes in each prediction sample by taking the preset survival time as a standard; determining an accuracy of the STAB1 in each prediction sample according to the first quantity and the second quantity. Wherein the prediction samples comprise target samples and validation samples. Wherein, the accuracy of the STAB1 in each prediction sample is shown in Table 1:

TABLE 1

In table 1, TCGA CN-AML (79) is a target sample, and the other four groups are prediction samples, and taking the target sample as an example, when calculating the accuracy of STAB1 in the target sample, specifically: (35+ 22)/79-0.72.

Then, the accuracy of STAB1 in other verification samples is calculated by the same method, and as can be seen from Table 1, the accuracy of STAB1 in prediction samples is more than 60%, which proves that the accuracy of STAB1 is feasible.

Furthermore, the feasibility of STAB1 was also predicted from the KM survival curve of STAB1 in each prediction sample, wherein the KM survival curve of the STAB1 gene in TCGA CN-AML sample is shown in FIG. 3, the KM survival curve of the STAB1 gene in GSE12417A (79) sample is shown in FIG. 4, the KM survival curve of the STAB1 gene in GSE71014(104) sample is shown in FIG. 5, and the KM survival curve of the STAB1 gene in GSE6891(187) sample is shown in FIG. 6. In fig. 3, 4, 5 and 6, the upper curve represents the KM survival curve corresponding to the low-expression STAB1, the lower curve represents the KM survival curve corresponding to the high-expression STAB1, and the corresponding n represents the number of low-expression samples and the number of high-expression samples, respectively.

Fig. 3, 4, 5, and 6 show the determination of the number of low expression samples and the number of high expression samples from the median value of the STAB 1. While the median value of STAB1 was also used to determine the high expression group of STAB1 and the low expression group of STAB1 in Table 1, it is statistically counted that the number of samples with survival time greater than 2 years in the low expression samples of STAB1 and the number of samples with survival time less than 2 years in the high expression samples of STAB1 are inconsistent.

As can be seen from fig. 3, 4, 5 and 6, the P value of the STAB1 gene was less than 0.05 in all of the four prediction samples, which further demonstrates that the accuracy of the STAB1 gene is feasible.

The CN-AML patients can then be stratified according to their gene expression levels of their target evaluation genes. When the gene expression level of the CN-AML patient target evaluation gene is greater than the median, determining that the CN-AML patient target evaluation gene is a high-expression gene, and classifying the current CN-AML patient into a poor prognosis group; prognostic survival time for poor prognosis group CN-AML patients is less than 2 years;

when the gene expression level of the target assessment gene of the CN-AML patient is less than the median, determining that the target assessment gene of the CN-AML patient is a low-expression gene, and dividing the current CN-AML patient into a good prognosis group; prognostic survival time of good prognosis group CN-AML patients was greater than 2 years.

After CN-AML patients are divided into a good prognosis group and a bad prognosis group according to the target evaluation gene expression quantity, corresponding medicine information can be indicated for CN-AML patients in the bad prognosis group so as to assist the clinical designation of an accurate layered treatment scheme and avoid delaying the treatment opportunity.

Meanwhile, the STAB1 high-expression patients are sensitive to small molecule inhibitors NVP-BHG712, GSK-J4, BRD-K30748066 and Tozasertib (Tozasertib), which indicates that the medicines possibly have certain effects in the STAB1 high-expression patients. Corresponding drug information is indicated for patients with high expression of STAB1, so that accurate layered treatment schemes can be clinically specified in an auxiliary mode, and treatment opportunity delay is avoided.

The method and the device for indicating the acute myeloid leukemia medicament provided by the embodiment of the invention have the following beneficial effects that:

the embodiment of the invention provides a method and a device for drug indication of acute myeloid leukemia, wherein the method comprises the following steps: obtaining a target evaluation gene of a normal karyotype acute myelogenous leukemia CN-AML patient, wherein the target evaluation gene is a STAB1 gene; the CN-AML patients are stratified according to the gene expression quantity of the target evaluation gene of the CN-AML patients, and the CN-AML patients are divided into a good prognosis group and a bad prognosis group; indicating corresponding drug information for CN-AML patients in the poor prognosis group; thus, screening the differential expression genes according to the screened differential expression genes in the target sample to obtain a plurality of prognosis-related differential expression genes related to survival time, screening the plurality of prognosis genes by combining clinical information, verifying each prognosis gene by utilizing a plurality of groups of sample data in a verification sample set to determine a target evaluation gene, dividing the CN-AML patients into good prognosis groups and poor prognosis groups according to the target evaluation genes of the CN-AML patients, and determining disease stratification of the patients; then, according to corresponding medicine information indicated by CN-AML patients with poor prognosis groups, accurate and layered treatment schemes are rapidly formulated in an auxiliary clinical mode, and treatment opportunity is prevented from being delayed; here, since only one target evaluation gene is determined, it is possible to easily perform prognostic evaluation and disease stratification for CN-AML patients; in addition, the target evaluation gene is the STAB1 gene, and the STAB1 gene is a membrane protein gene, so that the rapid detection can be easily carried out by using the existing RT-PCR or flow cytometry, the detection efficiency is improved, and the simplicity of the prognosis layering process is further improved.

The algorithms and displays presented herein are not inherently related to any particular computer, virtual machine, or other apparatus. Various general purpose systems may also be used with the teachings herein. The required structure for constructing such a system will be apparent from the description above. Moreover, the present invention is not directed to any particular programming language. It is appreciated that a variety of programming languages may be used to implement the teachings of the present invention as described herein, and any descriptions of specific languages are provided above to disclose the best mode of the invention.

In the description provided herein, numerous specific details are set forth. It is understood, however, that embodiments of the invention may be practiced without these specific details. In some instances, well-known methods, structures and techniques have not been shown in detail in order not to obscure an understanding of this description.

Similarly, it should be appreciated that in the foregoing description of exemplary embodiments of the invention, various features of the invention are sometimes grouped together in a single embodiment, figure, or description thereof for the purpose of streamlining the disclosure and aiding in the understanding of one or more of the various inventive aspects. However, the disclosed method should not be interpreted as reflecting an intention that: that the invention as claimed requires more features than are expressly recited in each claim. Rather, as the following claims reflect, inventive aspects lie in less than all features of a single foregoing disclosed embodiment. Thus, the claims following the detailed description are hereby expressly incorporated into this detailed description, with each claim standing on its own as a separate embodiment of this invention.

Those skilled in the art will appreciate that the modules in the device in an embodiment may be adaptively changed and disposed in one or more devices different from the embodiment. The modules or units or components of the embodiments may be combined into one module or unit or component, and furthermore they may be divided into a plurality of sub-modules or sub-units or sub-components. All of the features disclosed in this specification (including any accompanying claims, abstract and drawings), and all of the processes or elements of any method or apparatus so disclosed, may be combined in any combination, except combinations where at least some of such features and/or processes or elements are mutually exclusive. Each feature disclosed in this specification (including any accompanying claims, abstract and drawings) may be replaced by alternative features serving the same, equivalent or similar purpose, unless expressly stated otherwise.

Furthermore, those skilled in the art will appreciate that while some embodiments herein include some features included in other embodiments, rather than other features, combinations of features of different embodiments are meant to be within the scope of the invention and form different embodiments. For example, in the following claims, any of the claimed embodiments may be used in any combination.

The various component embodiments of the invention may be implemented in hardware, or in software modules running on one or more processors, or in a combination thereof. Those skilled in the art will appreciate that a microprocessor or Digital Signal Processor (DSP) may be used in practice to implement some or all of the functionality of some or all of the components of a gateway, proxy server, system according to embodiments of the present invention. The present invention may also be embodied as apparatus or device programs (e.g., computer programs and computer program products) for performing a portion or all of the methods described herein. Such programs implementing the present invention may be stored on computer-readable media or may be in the form of one or more signals. Such a signal may be downloaded from an internet website or provided on a carrier signal or in any other form.

It should be noted that the above-mentioned embodiments illustrate rather than limit the invention, and that those skilled in the art will be able to design alternative embodiments without departing from the scope of the appended claims. In the claims, any reference signs placed between parentheses shall not be construed as limiting the claim. The word "comprising" does not exclude the presence of elements or steps not listed in a claim. The word "a" or "an" preceding an element does not exclude the presence of a plurality of such elements. The invention may be implemented by means of hardware comprising several distinct elements, and by means of a suitably programmed computer. In the unit claims enumerating several means, several of these means may be embodied by one and the same item of hardware. The usage of the words first, second and third, etcetera do not indicate any ordering. These words may be interpreted as names.

The above description is only exemplary of the present invention and should not be taken as limiting the scope of the present invention, and any modifications, equivalents, improvements, etc. that are within the spirit and principle of the present invention should be included in the present invention.

Claims

1. A method for drug indication of acute myeloid leukemia, comprising:

2. The method of claim 1, wherein the stratifying the prognostic survival time of CN-AML patients based on gene expression levels of target assessment genes for said CN-AML patients comprises:

determining the median of the expression quantity of each gene;

3. The method of claim 1, wherein said indicating respective drug information to CN-AML patients in a poor prognosis group comprises:

4. The method of claim 1, wherein said indicating respective drug information to CN-AML patients in a poor prognosis group comprises:

5. The method of claim 1, wherein the method comprises:

6. The method of claim 5, wherein obtaining gene expression information of the target sample from the database comprises:

7. The method of claim 5, wherein the screening the genes of the first type of sample and the second type of sample according to a predetermined first screening condition to obtain a plurality of differentially expressed genes comprises:

8. The method of claim 5, wherein the screening the plurality of differentially expressed genes according to a predetermined second screening condition to obtain a plurality of differentially expressed genes associated with prognosis with respect to the survival time comprises:

9. The method according to claim 5, wherein the performing a multi-factor proportional risk Cox regression analysis on the plurality of prognostically relevant differentially expressed genes based on clinical factors affecting the prognosis of CN-AML and prognostic factors affecting AML survival to obtain independent prognostic genes comprises:

10. A device for drug indication of acute myeloid leukemia, said device comprising: