CN113913528A - Method for predicting sensitivity of melanoma patient to drug MAGE-A3 - Google Patents

Abstract

The invention discloses a method for predicting the sensitivity of a melanoma patient to a drug MAGE-A3, which comprises a biomarker capable of predicting the sensitivity of the melanoma patient to the drug MAGE-A3, wherein the biomarker is UTY, SLITRK6 and/or LONRF2, has higher accuracy for predicting the sensitivity of the melanoma patient to the drug MAGE-A3, can assist a clinician in selecting a proper immunotherapy method, and has excellent application prospect in clinic.

Description

Method for predicting sensitivity of melanoma patient to drug MAGE-A3

Technical Field

The invention belongs to the technical field of biomedicine, and particularly relates to a method for predicting the sensitivity of a melanoma patient to a drug MAGE-A3 by using a biomarker.

Background

Melanoma (Melanoma) is a highly malignant tumor derived from melanocytes of the basal layer of the epidermis, originating from melanocytes of the nerve ridge, which preferably originates in the skin or mucous membranes adjacent to the skin, and is the most aggressive and lethal cancer of the skin. In recent years, the incidence and mortality of melanoma have a significant increasing trend worldwide, accounting for 1% -3% of all malignant tumors, but the incidence rate is increasing year by year at a growth rate of 6% -7%, and the incidence age is a trend of younger, which is one of the most rapidly growing malignant tumors. The incidence of melanoma is quite obvious in ethnic difference, the highest incidence is found in caucasians, and the lower incidence is found in yellow and black. The main cause of death in melanoma patients is the propensity of the tumor to metastasize rapidly. Melanoma has the characteristics of high malignancy, rapid progress, early metastasis and the like, and in recent years, the clinical history of melanoma is revolutionarily changed due to the rapid development of immunotherapy and targeted therapy, the therapy has certain effect, and the survival period of melanoma patients can be remarkably prolonged, but the therapy is limited by the practical clinical application, and no satisfactory therapy is available at present for the melanoma patients with advanced tumor metastasis.

Therefore, further intensive studies are urgently needed for the research on the pathogenesis of melanoma. Currently, there is increasing evidence that melanoma is pathologically formed involving multiple genes, multiple signaling pathways, and complex changes in gene regulation. Therefore, identifying some potential target molecules in melanoma progression, further exploring the relevant mechanisms of melanoma occurrence and development, searching for new tumor markers and treating targets to prolong the life cycle of patients become a problem to be solved urgently. Currently, the immuno-drug MAGE-A3 treatment protocol is widely applied in the adjuvant therapy of clinical melanoma, but it is still difficult to predict which melanoma patients will be sensitive and/or reactive to the immuno-drug MAGE-A3 treatment protocol, and predicting the sensitivity or reactivity of melanoma patients to the immuno-drug MAGE-A3 remains a huge challenge, and therefore, it is important to the art to find biomarkers that can be used for accurate prediction of the sensitivity and/or reactivity of melanoma patients to the immuno-drug MAGE-A3 treatment protocol, and to apply them in the screening of melanoma patients before treatment, which can help to realize individualized treatment.

Disclosure of Invention

In view of the above, in order to solve the above technical drawbacks existing in the prior art, the present invention aims to provide a method for predicting the sensitivity of melanoma patients to the drug MAGE-a 3.

The above object of the present invention is achieved by the following technical solutions:

in a first aspect, the present invention provides the use of a reagent for detecting the level of expression of a biomarker in the manufacture of a product for predicting the sensitivity and/or reactivity of melanoma to the immunopharmaceutical MAGE-a 3.

Further, the biomarker is UTY, slicrk 6, and/or LONRF 2.

Further, the detailed information of the biomarkers UTY, slicrk 6 and LONRF2 of the present invention is as follows:

gene UTY (ubiquitin transformed tetratricopeptide repeat association, Y-linked): gene ID 7404, specific position Yq11.221;

gene slicrk 6(SLIT and NTRK like family member 6): gene ID is 84189, with a specific position of 13q 31.1;

gene LONRF2(LON peptidase N-terminal domain and ring finger 2): gene ID is 164832, with a specific position of 2q 11.2.

Further, the agent is selected from:

a probe that specifically recognizes the biomarker; or

Primers that specifically amplify the biomarkers; or

Antibodies, antibody fragments, affinity proteins that specifically bind to the biomarkers.

Further, the product comprises a reagent for detecting the expression level of the biomarker by a nucleic acid hybridization technology, a nucleic acid amplification technology, a protein immunity technology, a sequencing technology, a chromatography technology and a mass spectrometry technology.

Further, the nucleic acid hybridization technique refers to a process in which complementary nucleotide sequences (DNA to DNA, DNA to RNA, RNA to RNA, etc.) form noncovalent bonds through Watson-Crick base pairing, thereby forming stable homoduplex or heteroduplex molecules, which is also called nucleic acid hybridization.

Further, the nucleic acid amplification technology is a generic name of a large class of technical methods, and the existing nucleic acid amplification technology comprises conventional PCR, real-time fluorescence PCR, isothermal nucleic acid amplification technology and the like, can specifically amplify a trace amount of target DNA by millions of times, thereby greatly improving the analysis and detection capability of DNA molecules, detecting single-molecule DNA or samples containing only 1 target DNA molecule in every 10 ten thousand cells, and having the advantages of high sensitivity, strong specificity, rapidness, simplicity and the like.

Further, the protein immunization technique refers to a generic term for a class of methods including radioimmunoassay, direct, indirect or contrast enzyme-linked immunosorbent assay, enzyme immunoassay, fluorescence immunoassay, western blotting, immunoprecipitation, any particle-based immunoassay (e.g., using gold, silver or latex particles, magnetic particles or quantum dots) for the detection of a target, and the protein immunization can be performed in the form of a microtiter plate or strip.

Further, the sequencing technology includes (but is not limited to) first-generation sequencing, second-generation sequencing, and third-generation sequencing, wherein the first-generation sequencing is also called Sanger sequencing, and is a sequencing technology utilizing a DNA polymerase synthesis reaction, and the first-generation sequencing is a sequencing technology based on the Sanger method; the second generation sequencing is based on massively parallel sequencing technology (MPS), and can simultaneously complete the synthesis of a complementary strand of a sequencing template and the acquisition of sequence data; the third generation sequencing is based on single molecule sequencing and massively parallel sequencing technology.

Further, the chromatographic technique refers to a method for separating and analyzing each component in a complex mixture, and utilizes different substances having different distribution coefficients in a system composed of a stationary phase and a mobile phase, and when the two phases move relatively, the substances move together with the mobile phase and are repeatedly distributed between the two phases, so that each substance is separated.

Further, the mass spectrometry technique is a method of separating and detecting moving ions (charged atoms, molecules or molecular fragments, molecular ions, isotope ions, fragment ions, rearrangement ions, multiply-charged ions, metastable ions, negative ions, and ions generated by ion-molecule interaction) according to their mass-to-charge ratios using an electric field and a magnetic field. The accurate mass of the ions is measured, and the compound composition of the ions can be determined.

Furthermore, the product comprises a kit, a chip, test paper and a high-throughput sequencing platform.

Further, in a specific embodiment of the present invention, the melanoma is metastatic melanoma.

In a second aspect of the invention there is provided a product for predicting the sensitivity and/or reactivity of a melanoma patient to the immunopharmaceutical MAGE-a 3.

Further, the product comprises an agent that detects the level of expression of the biomarkers UTY, SLITRK6, and/or LONRF2 in the sample.

Further, the reagent includes a reagent for detecting the mRNA expression level of the biomarker, and a reagent for detecting the protein expression level of the biomarker.

Further, the reagents include probes that specifically recognize the biomarkers, primers that specifically amplify the biomarkers, or antibodies, antibody fragments, affinity proteins that specifically bind to the biomarkers.

Furthermore, the product comprises a kit, a chip, test paper and a high-throughput sequencing platform.

In a particular embodiment of the invention, the sample is selected from blood or tissue.

A third aspect of the invention provides a system/apparatus for predicting the sensitivity and/or reactivity of a melanoma patient to the immune drug MAGE-a 3.

Further, the system/apparatus includes:

(1) an analysis unit: the analysis unit is for detecting the expression level of the biomarkers UTY, SLITRK6, and/or LONRF2 in a sample of a subject;

(2) an evaluation unit: the evaluation unit comprises a stored reference and a data processor having implemented an algorithm for comparing the expression levels of the biomarkers UTY, SLITRK6, and/or LONRF2 detected by the analysis unit with the stored reference, thereby predicting the sensitivity and/or reactivity of melanoma patients to the immunological drug MAGE-A3;

(3) an evaluation result output unit: the evaluation result output unit outputs the prediction result of the melanoma patient sensitivity and/or reactivity to the immuno-drug MAGE-A3 obtained by the evaluation unit.

A fourth aspect of the invention provides the use of the biomarkers UTY, SLITRK6, and/or LONRF2 in the construction of a system/device for predicting the sensitivity and/or reactivity of a melanoma patient to the immune drug MAGE-A3.

Further, in particular embodiments of the invention, the invention assesses the diagnostic efficacy or accuracy of the biomarker sensitivity and/or reactivity to treatment with the immuno-drug MAGE-a3 in melanoma patients by the area under the subject's working characteristic curve (AUC). The accuracy of a diagnostic method is best described by its Receiver Operating Characteristics (ROC). ROC plots are line graphs of all sensitivity/specificity pairs derived from continuously varying decision thresholds across the entire data range observed. In one embodiment of the present invention, the diagnostic efficacy of the biomarkers of the present invention in predicting the sensitivity or reactivity of a melanoma patient to treatment with the immunopharmaceutical MAGE-A3 can be evaluated based on the area under the working characteristic curve (AUC) of the subject, wherein if the AUC > 0.6 indicates that the biomarkers have a certain diagnostic efficacy, the biomarkers can be used as markers for predicting the sensitivity or reactivity of a melanoma patient to treatment with the immunopharmaceutical MAGE-A3; if AUC > 0.7, it indicates that the biomarker has relatively good diagnostic efficacy, and can be used as a marker for predicting the sensitivity or reactivity of melanoma patients to the treatment with the immune drug MAGE-A3; if AUC > 0.8, it indicates that the biomarker has better diagnostic efficacy and can be used as a marker for predicting the sensitivity or responsiveness of melanoma patients to the treatment with the immuno-drug MAGE-A3.

The biomarkers described herein include the biomarkers, as well as their encoded proteins and homologs, mutations, and isoforms thereof, which encompass full-length unprocessed biomarkers, as well as any form of biomarker that results from processing in a cell, as well as naturally occurring variants (e.g., splice variants or allelic variants) of the biomarkers. The gene ID is available at https:// www.ncbi.nlm.nih.gov/gene/.

In order to further clarify the content of the present invention, some of the scientific terms referred to in the present invention are explained as follows.

"reactive" as used herein, means that a patient/subject having, suspected of having, or predisposed to having melanoma, exhibits a response to treatment with the immuno-drug MAGE-a 3. One skilled in the art will readily determine whether a patient/subject employing the immunotherapeutic MAGE-a3 treatment regimen will show a response according to the methods described herein.

"sensitive" as used herein, means that a patient/subject having, suspected of having, or predisposed to having melanoma, in some manner, shows a positive response to treatment with the immuno-drug MAGE-a 3. One skilled in the art will readily determine whether a patient/subject employing the immunotherapeutic MAGE-a3 treatment regimen shows a positive response according to the methods described herein.

As used herein, "biomarker," as well as "marker" and "marker molecule," refers to an indicator of a patient's phenotype, and in particular embodiments of the present invention, refers to a marker that is differentially expressed between melanoma patients (responders) sensitive or responsive to an immunotherapeutic MAGE-A3 treatment regimen and melanoma patients (non-responders) not sensitive or responsive to an immunotherapeutic MAGE-A3 treatment regimen, including, but not limited to: genes, proteins, small molecule metabolites, carbohydrates, glycolipid-based molecules, and the like. A biomarker may be DNA comprising all or part of a nucleic acid sequence encoding the biomarker, or the complement of such a sequence. Biomarker nucleic acids useful in the present invention are considered to include DNA and RNA comprising all or part of any nucleic acid sequence of interest.

The "sample" as used herein, as with the "sample", "subject sample", and "melanoma patient sample", may be a cell or tissue sample (e.g., biopsy), a biological fluid, or an extract (e.g., a protein or DNA extract obtained from a subject). In particular, the sample may be a tumor sample, e.g. a solid tumor, e.g. a melanoma tissue sample. The sample may be a sample freshly obtained from the subject, or may be a sample that has been processed and/or stored (e.g., frozen, fixed, or subjected to one or more purification, enrichment, or extraction steps) prior to the determination being made, such as treated with a reagent, stabilized, or enriched for certain components (such as proteins or polynucleotides), or embedded in a semi-solid or solid matrix for sectioning purposes. Samples described in the present invention include, but are not limited to: tissue, whole blood, blood-derived cells, serum, plasma, lymph, synovial fluid, cell extracts and combinations thereof.

The expression level of the biomarker of the present invention refers to the expression level of the biomarker in a sample of a subject. Determining the level of gene expression may involve determining the level of protein expressed by the gene in a sample comprising cancer cells obtained from the individual. Protein expression levels can be determined by any useful means, including the use of immunoassays. For example, expression levels can be determined by Immunohistochemistry (IHC), western blotting, ELISA, immunoelectrophoresis, immunoprecipitation, flow cytometry, Mass cytometry, and immunostaining. Using any of these methods, the relative expression levels of the proteins of the biomarkers disclosed herein can be determined.

As an alternative embodiment, the expression level of the gene may also be detected using advanced sequencing methods. For example, Illumina can be used to detect biomarkers. Next generation Sequencing (e.g., Sequencing-By-Synthesis or TruSeq methods using, for example, the HiSeq, HiScan, genome Analyzer, or MiSeq systems). Biomarkers can also be detected using ion beam sequencing or other suitable semiconductor sequencing methods.

As an alternative embodiment, RNase profiling (Mapping) can be used to quantify biomarkers using mass spectrometry. The isolated RNA may be enzymatically digested with an RNA endonuclease (RNase) having high specificity (e.g., RNase T1, which cleaves 3' to all unmodified guanosine residues) prior to analysis of the isolated RNA by MS or tandem MS (MS/MS) methods. The first method developed used reverse phase HPLC coupled directly to ESI-MS to perform on-line chromatographic separation of endonuclease digests. The presence of post-transcriptional modifications can be revealed by mass shifts from those expected based on the RNA sequence. Ions of abnormal mass/charge values can then be isolated for tandem MS sequencing, thereby locating the sequence position of the post-transcriptionally modified nucleoside.

Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) has also been used as an analytical method to obtain information about post-transcriptionally modified nucleosides. MALDI-based methods can be distinguished from ESI-based methods by separation steps. In MALDI-MS, mass spectrometry is used to separate biomarkers.

"primer" as used herein refers to a nucleic acid sequence having a short free 3' -hydroxyl group, which is a short nucleic acid that can form a base pair with a complementary template and serves as an origin of replication of the template strand. The primers can prime DNA synthesis in the presence of reagents for polymerization (i.e., DNA polymerase or reverse transcriptase) and four different nucleoside triphosphates in appropriate buffer solutions and temperatures. The PCR conditions and the lengths of the sense and antisense primers can be appropriately selected according to the techniques known in the art.

The "probe" as used herein refers to a nucleic acid fragment (e.g., RNA or DNA) corresponding to several bases to several hundred bases that can specifically bind to mRNA, and can confirm the presence or absence and expression level of a specific mRNA by a tag. The probe may be prepared in the form of an oligonucleotide probe, a single-stranded DNA probe, a double-stranded DNA probe, or an RNA probe. Suitable probes and hybridization conditions may be appropriately selected according to techniques known in the art.

The term "antibody" as used herein is well known in the art and refers to an immunoglobulin specific for an antigenic site. The antibody of the present invention refers to an antibody that specifically binds to the biomarker protein of the present invention, and can be produced according to a conventional method in the art. Forms of antibodies include polyclonal or monoclonal antibodies, antibody fragments (such as Fab, Fab ', F (ab') 2, and Fv fragments), single chain Fv (scfv) antibodies, multispecific antibodies (such as bispecific antibodies), monospecific antibodies, monovalent antibodies, chimeric antibodies, humanized antibodies, human antibodies, fusion proteins comprising an antigen binding site of an antibody, and any other modified immunoglobulin molecule comprising an antigen binding site, so long as the antibody exhibits the desired biological binding activity.

The "peptide" and "polypeptide" as used herein mean a peptide having a high binding ability to a target substance and being not denatured during heat/chemical treatment. Also, due to its small size, it can be used as a fusion protein by attaching it to other proteins. In particular, since it can be specifically attached to a high molecular protein chain, it can be used as a diagnostic kit and a drug delivery substance.

The immune medicine MAGE-A3, like the immune therapeutic medicine MAGE-A3, the medicine MAGE-A3 and the medicine MAGE-A3, in the invention refers to a medicine for the immune therapy of metastatic melanoma, and the medicine specifically refers to a substance obtained by dissolving recombinant MAGE-A3 protein in an immune stimulant (AS02B or AS 15). The "MAGE-A3 immunotherapy", AS well AS "MAGE-A3 immunotherapy", AS used herein, refers to an immunotherapy method in which a substance obtained by dissolving a recombinant MAGE-A3 protein in an immunostimulant (AS02B or AS15) is administered to a patient with metastatic melanoma.

Compared with the prior art, the invention has the advantages and beneficial effects that:

the invention discovers for the first time that the combination of UTY, SLITRK6 and/or LONRF2 can be used for predicting the sensitivity or the reactivity of a melanoma patient to the treatment of an immune medicament MAGE-A3, has higher accuracy and higher AUC value, can assist a clinician to judge the sensitivity or the reactivity of the melanoma subject to the treatment of the immune medicament MAGE-A3, further realizes individualized treatment, and has very good clinical application prospect.

Drawings

Embodiments of the invention are described in detail below with reference to the attached drawing figures, wherein:

FIG. 1 is a graph showing the results of UTY differentially expressed between melanoma patients in the group responsive to treatment with the immunopharmaceutical MAGE-A3 and the group non-responsive to treatment with the immunopharmaceutical MAGE-A3;

figure 2 shows a graph of the results of differential expression of SLITRK6 between melanoma patients in the group responsive to treatment with the immunopharmaceutical MAGE-A3 and the group non-responsive to treatment with the immunopharmaceutical MAGE-A3;

FIG. 3 is a graph showing the results of the differential expression of LONRF2 between melanoma patients in the group responsive to treatment with the immunopharmaceutical MAGE-A3 and the group non-responsive to treatment with the immunopharmaceutical MAGE-A3;

FIG. 4 is a graph showing the results of UTY's diagnostic efficacy in predicting melanoma sensitivity to the immunopharmaceutical MAGE-A3;

FIG. 5 is a graph showing the results of SLITRK6 on the diagnosis efficacy in predicting melanoma sensitivity to the immunopharmaceutical MAGE-A3;

FIG. 6 is a graph showing the results of the diagnostic efficacy of LONRF2 for predicting melanoma sensitivity to the immunopharmaceutical MAGE-A3;

FIG. 7 is a graph showing the results of the diagnostic efficacy of the UTY + SLITRK6 combination on predicting melanoma sensitivity to the immunopharmaceutical MAGE-A3;

FIG. 8 is a graph showing the results of the diagnostic efficacy of the UTY + LONRF2 combination on predicting melanoma sensitivity to the immunopharmaceutical MAGE-A3;

FIG. 9 is a graph showing the results of the diagnostic efficacy of SLITRK6+ LONRF2 in combination on predicting the sensitivity of melanoma to the immunopharmaceutical MAGE-A3;

FIG. 10 is a graph showing the results of the diagnostic efficacy of UTY + SLITRK6+ LONRF2 in combination for predicting the sensitivity of melanoma to the immunopharmaceutical MAGE-A3.

Detailed Description

The present invention is further illustrated by the following examples, which should be construed as being merely illustrative and not limitative of the remainder of the disclosure. As will be understood by those of ordinary skill in the art: many changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention. The following examples are examples of experimental methods not indicating specific conditions, and the detection is usually carried out according to conventional conditions or according to the conditions recommended by the manufacturers. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs, and any methods and materials similar or equivalent to those described herein may be applied.

Example 1 screening 1, data sources and screening methods for genes associated with sensitivity of metastatic melanoma to the immunopharmaceutical MAGE-A3

Patients clinically diagnosed with metastatic melanoma were collected and were not treated with the immuno-drug MAGE-A3 prior to receiving the immuno-drug MAGE-A3. Before the patients with metastatic melanoma collected as described above were treated with the immuno-drug MAGE-A3 immunotherapy, tumor biopsies of the patients were collected and genes differentially expressed in tumor tissues of patients with response (R, responder) or non-response (NR, non-responder) groups after the MAGE-A3 immunotherapy were examined. The MAGE-A3 immunotherapy refers to the administration of an immune drug MAGE-A3 to a patient with metastatic melanoma, and the immune drug MAGE-A3 refers to a substance obtained by dissolving recombinant MAGE-A3 protein in an immunostimulant (AS02B or AS 15). Wherein, according to whether the metastatic melanoma is responsive/sensitive to the immune drug MAGE-A3, the melanoma patients are divided into a response group (R, responder) and a non-response group (NR, non-responder) according to RECIST 1.0 (standard for evaluating the curative effect of solid tumors); the reactive group comprises melanoma patients with Complete Response (CR) and melanoma patients with Partial Response (PR), and the non-reactive group comprises Stable Disease (SD) (> 4 months) and Progressive Disease (PD); the data from this study are derived from GSE35640, and the proportion of metastatic melanoma patients who were non-responsive and reactive to the immunopharmaceutical MAGE-a3 was as follows: non-responder: reposder-34: 22;

comparing the genes differentially expressed between the metastatic melanoma patients of the response group and the metastatic melanoma patients of the non-response group; analysis of the differentially expressed genes was performed using the "limma" package (version 3.36.5) in the R software, in which,screening criteria for differentially expressed genes was p<0.01，|log₂FC|>1。

2. Results of the experiment

The results show that among the differentially expressed genes obtained by screening, UTY, SLITRK6 and LONRF2 showed significant differential expression between patients with metastatic melanoma in the group responsive to the treatment of the immune drug MAGE-A3 and the group non-responsive to the treatment of the immune drug MAGE-A3, and the expression trends of UTY, SLITRK6 and LONRF2 in the response group were down-regulated, down-regulated and down-regulated respectively (see FIGS. 1-3), and the results suggest that UTY, TRSLITK 6 and LONRF2 are potential biomarkers for predicting the treatment sensitivity of melanoma to the treatment of the immune drug MAGE-A3.

Example 2 application of differential expression gene obtained by screening in predicting melanoma sensitivity to treatment with immune drug MAGE-A3

1. Experimental methods

Performing Receiver Operating Characteristic (ROC) analysis on the genes differentially expressed between the response group and the non-response group screened in example 1 using package R "pROC" (version 1.15.0), calculating the area under the operating characteristic curve (AUC) of the subject to evaluate the accuracy of a single differentially expressed gene, any two differentially expressed gene combinations, and three differentially expressed gene combinations, respectively, for predicting the sensitivity of melanoma to treatment with the immuno-drug MAGE-A3, wherein the AUC values range from 0 to 1;

wherein, when judging the diagnosis effectiveness of the single differential expression gene for predicting the treatment sensitivity of melanoma on the immune drug MAGE-A3, directly adopting the expression quantity of the single differential expression gene for analysis, selecting the level corresponding to the maximum Youden index as the cutoff value, and the differential expression gene with AUC of 0.5< AUC <0.8 is used for joint analysis;

when the diagnosis effectiveness of the combination of any two differential expression genes and the combination of three differential expression genes on the prediction of the treatment sensitivity of melanoma to the immune medicament MAGE-A3 is judged, Logitics regression analysis is carried out on the expression level of each differential expression gene, the probability of each melanoma patient being sensitive to the treatment of the immune medicament MAGE-A3 is calculated through a fitted regression curve, different probability division thresholds are determined, the accuracy, the specificity, the sensitivity and the like of the combination of any two differential expression genes and the combination of three differential expression genes on the prediction of the treatment sensitivity of melanoma to the immune medicament MAGE-A3 are calculated according to the determined probability division thresholds.

2. Results of the experiment

The results show that the diagnosis efficacy of the gene combination UTY + SLITRK6, UTY + LONRF2, SLITRK6+ LONRF2 and UTY + SLITRK6+ LONRF2 on prediction of the treatment sensitivity of melanoma to the immune medicament MAGE-A3 is obviously superior to that of a single gene, and the AUC values of the prediction of the treatment sensitivity of melanoma to the immune medicament MAGE-A3 by using the gene combination are high (see table 1 and figure 4-figure 10), which shows that the genes UTY, TRSLIK 6 and/or LONRF2 can be used for clinically assisting doctors to judge the sensitivity and/or the reactivity of melanoma to the treatment of the immune medicament MAGE-A3 so as to guide the clinicians to select effective treatment methods, realize accurate individualized treatment and have very good clinical application prospects.

TABLE 1 diagnostic potency of genes for predicting melanoma sensitivity to treatment with the immunopharmaceutical MAGE-A3

Gene	AUC value
		UTY	0.684
SLITRK6	0.699
		LONRF2	0.742
UTY+SLITRK6	0.832
		UTY+LONRF2	0.853
SLITRK6+LONRF2	0.838
		UTY+SLITRK6+LONRF2	0.912

The above description of the embodiments is only intended to illustrate the method of the invention and its core idea. It should be noted that, for those skilled in the art, without departing from the principle of the present invention, several improvements and modifications can be made to the present invention, and these improvements and modifications will also fall into the protection scope of the claims of the present invention.

Claims (10)

1. Use of a reagent for detecting the level of expression of a biomarker, wherein the biomarker is UTY, slicrk 6, and/or LONRF2, in the manufacture of a product for predicting the sensitivity and/or reactivity of melanoma to the immunopharmaceutical MAGE-a 3.

2. The use according to claim 1, wherein the agent is selected from the group consisting of:

a probe that specifically recognizes the biomarker; or

Primers that specifically amplify the biomarkers; or

Antibodies, antibody fragments, affinity proteins that specifically bind to the biomarkers.

3. The use according to claim 1, wherein the product comprises reagents for detecting the level of expression of the biomarkers by nucleic acid hybridization techniques, nucleic acid amplification techniques, protein immunization techniques, sequencing techniques, chromatography techniques, mass spectrometry techniques.

4. The use of claim 1, wherein the product comprises a kit, a chip, a strip, a high throughput sequencing platform.

5. A product for predicting the sensitivity and/or reactivity of a melanoma patient to the immunological drug MAGE-a3, said product comprising reagents for detecting the level of expression of the biomarkers UTY, SLITRK6, and/or LONRF2 in a sample.

6. The product of claim 5, wherein the reagents comprise a reagent that detects the mRNA expression level of the biomarker, a reagent that detects the protein expression level of the biomarker.

7. The product of claim 6, wherein the reagents comprise probes that specifically recognize the biomarkers, primers that specifically amplify the biomarkers, or antibodies, antibody fragments, affinity proteins that specifically bind to the biomarkers.

8. The product of claim 5, wherein the product comprises a kit, a chip, a strip, a high throughput sequencing platform.

9. A system/device for predicting the sensitivity and/or reactivity of melanoma patients to the immune drug MAGE-a3, the system/device comprising:

(1) an analysis unit: the analysis unit is for detecting the expression level of the biomarkers UTY, SLITRK6, and/or LONRF2 in a sample of a subject;

(2) an evaluation unit: the evaluation unit comprises a stored reference and a data processor having implemented an algorithm for comparing the expression levels of the biomarkers UTY, SLITRK6, and/or LONRF2 detected by the analysis unit with the stored reference, thereby predicting the sensitivity and/or reactivity of melanoma patients to the immunological drug MAGE-A3;

(3) an evaluation result output unit: the evaluation result output unit outputs the prediction result of the melanoma patient sensitivity and/or reactivity to the immuno-drug MAGE-A3 obtained by the evaluation unit.

10. Use of the biomarkers UTY, SLITRK6, and/or LONRF2 in the construction of a system/device for predicting the sensitivity and/or reactivity of melanoma patients to the immunopharmaceutical MAGE-A3.

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