Method for predicting prognosis of skin cancer and its use

Abstract

Provided is a technique for predicting prognosis of skin cancer. Provided is a method for predicting prognosis of skin cancer including: a step for obtaining a correlation amount corelated to an expression amount of glucose-6-phosphate dehydrogenase in a specimen collected from a patient of skin cancer; and a step for determining poor prognosis of skin cancer when the correlation amount is large as compared with when the correlation amount is small.

Description

[Technical field]

[0001] The present disclosure relates to a method for predicting the prognosis of skin cancer. This application is based on Japanese Application No. 2020-072636 filed on April 15, 2020, the contents of which are incorporated herein by reference. [Background technology]

[0002] In recent years, anti-cancer therapy using immune checkpoint inhibition, including inhibitors of PD-L1 (programmed cell death ligand 1) and its receptor PD-1 (programmed cell death 1), has achieved excellent results in various cancers including malignant melanoma. Clinical use of anti-PD-L1 antibody drugs has also begun for Merkel cell carcinoma, a type of skin cancer. PD-L1 expressed in tumor cells binds to PD-1 expressed in T cells, inhibiting T cell activation and immune responses and helping tumor growth. For this reason, high expression of PD-L1 is known to indicate poor prognosis in many cancers. However, it is known that high expression of PD-L1 suggests good prognosis in Merkel cell carcinoma (e.g., Non-Patent Document 1).

[0003] It is also known that PD-L1 has heterogeneity even within the same case, and the present inventors were the first to discover and report that such heterogeneity also exists in Merkel cell carcinoma (Non-Patent Document 2). The present inventors were also the first to discover and report that no correlation was observed between the expression level of PD-L1 in the primary focus of Merkel cell carcinoma and prognosis, but that a strong correlation was observed between the expression level of PD-L1 in the skin metastatic focus of Merkel cell carcinoma and prognosis (Non-Patent Document 3). [Prior art documents] [Non-patent literature]

[0004] [Non-Patent Document 1] Lipson EJ, et al., Cancer Immunol. Res., 54-63, 2013 [Non-Patent Document 2] Nakamura M. et al., Br J Dermatol., 1228-1229, 2019 [Non-Patent Document 3] Nakamura M. et al., J Dermatol. Sci., 165-167, 2020 Summary of the Invention [Problem to be solved by the invention]

[0005] According to Non-Patent Document 2, the expression level of PD-L1 varies greatly depending on the time and site of resection, even in the same case. For this reason, it is difficult to predict the prognosis of Merkel cell carcinoma using PD-L1. In addition, according to Non-Patent Document 3, although a strong correlation is shown between the expression level of PD-L1 in skin metastasis of Merkel cell carcinoma and prognosis, it is difficult to use it clinically as a prognostic predictor if the prognosis can only be predicted after skin metastasis is recognized. For this reason, other methods for predicting the prognosis of Merkel cell carcinoma have been desired. Note that such a problem is not limited to Merkel cell carcinoma but is a common problem in other skin cancers. [Means for solving the problem]

[0006] The present invention can be realized in the following forms.

[0007] (1) According to one embodiment of the present invention, there is provided a method for predicting the prognosis of skin cancer. This method for predicting the prognosis of skin cancer includes the steps of: determining an amount of correlation that correlates with the expression level of glucose-6-phosphate dehydrogenase in a sample collected from a patient with skin cancer; and determining that the prognosis of the skin cancer is worse when the amount of correlation is high than when the amount of correlation is low. According to this method for predicting the prognosis of skin cancer, the prognosis of skin cancer can be predicted using the expression level of glucose-6-phosphate dehydrogenase in a sample collected from a patient with skin cancer as an index.

[0008] (2) In the above-mentioned method for predicting the prognosis of skin cancer, the skin cancer may be at least one selected from the group consisting of Merkel cell carcinoma, malignant melanoma, squamous cell carcinoma, extramammary Paget's disease, and cutaneous angiosarcoma.

[0009] (3) In the above-mentioned method for predicting the prognosis of skin cancer, the skin cancer may include Merkel cell carcinoma.

[0010] (4) In the above-described method for predicting the prognosis of skin cancer, the step of determining the correlation amount may include a step of determining the expression level of glucose-6-phosphate dehydrogenase by immunohistochemical staining of the sample. According to this method for predicting the prognosis of skin cancer, the prognosis of skin cancer can be easily predicted by determining the expression level of glucose-6-phosphate dehydrogenase by immunohistochemical staining of a sample collected from a skin cancer patient.

[0011] (5) In the above-described method for predicting the prognosis of skin cancer, the step of determining the correlation amount may include a step of determining the expression level of the mRNA encoding glucose-6-phosphate dehydrogenase in the sample. According to this method for predicting the prognosis of skin cancer, the prognosis of skin cancer can be easily predicted by determining the expression level of the mRNA encoding glucose-6-phosphate dehydrogenase in a sample collected from a skin cancer patient.

[0012] (6) In the above-described method for predicting the prognosis of skin cancer, the step of determining the correlation amount may include a step of measuring the activity of glucose-6-phosphate dehydrogenase in the blood or serum sample. According to this method for predicting the prognosis of skin cancer, the prognosis of skin cancer can be easily predicted by measuring the activity of glucose-6-phosphate dehydrogenase in the blood or serum sample collected from a patient with skin cancer.

[0013] (7) According to another aspect of the present invention, there is provided a method for evaluating the effectiveness of an immune checkpoint inhibitor against skin cancer. The evaluation method of this aspect includes the steps of: determining an amount of correlation that correlates with the expression level of glucose-6-phosphate dehydrogenase in a sample collected from a patient with skin cancer; and evaluating the effectiveness of the immune checkpoint inhibitor as being higher when the amount of correlation is low than when the amount of correlation is high. According to the evaluation method of this aspect, the effectiveness of the immune checkpoint inhibitor can be evaluated using the expression level of glucose-6-phosphate dehydrogenase in a sample collected from a patient with skin cancer as an index.

[0014] (8) According to another aspect of the present invention, there is provided a method for determining whether or not to administer an immune checkpoint inhibitor to a patient with skin cancer. The method of this aspect includes an evaluation method for evaluating the effectiveness of an immune checkpoint inhibitor against skin cancer, and includes a step of determining to administer or continue the administration of the immune checkpoint inhibitor to the patient when the effectiveness of the immune checkpoint inhibitor is evaluated to be high. According to the method of this aspect, the expression level of glucose-6-phosphate dehydrogenase in a sample collected from a patient with skin cancer can be used as an index to determine whether or not to administer an immune checkpoint inhibitor.

[0015] (9) According to another aspect of the present invention, there is provided a method for evaluating the malignancy of skin cancer. This method for evaluating the malignancy of skin cancer includes the steps of: determining an amount of correlation that correlates with the expression level of glucose-6-phosphate dehydrogenase in a sample collected from a skin cancer patient; and determining that the malignancy of the skin cancer is higher when the amount of correlation is high compared to when the amount of correlation is low. According to this method for evaluating the malignancy of skin cancer, the expression level of glucose-6-phosphate dehydrogenase in a sample collected from a skin cancer patient can be used as an index to evaluate the malignancy of skin cancer.

[0016] (10) According to another aspect of the present invention, there is provided a biomarker for predicting the prognosis of skin cancer, the biomarker comprising glucose-6-phosphate dehydrogenase in a sample collected from a patient with the skin cancer, wherein a high expression level of the biomarker in the sample indicates a poorer prognosis of the skin cancer than a low expression level.

[0017] (11) According to another aspect of the present invention, there is provided a biomarker for evaluating the efficacy of an immune checkpoint inhibitor against skin cancer, the biomarker comprising glucose-6-phosphate dehydrogenase in a sample collected from a patient with the skin cancer, and a low expression level of the biomarker in the sample indicates a higher efficacy of the immune checkpoint inhibitor than a high expression level.

[0018] (12) According to another aspect of the present invention, there is provided a biomarker for assessing the malignancy of skin cancer, the biomarker comprising glucose-6-phosphate dehydrogenase in a sample collected from a patient with the skin cancer, and a high expression level of the biomarker in the sample indicates a high malignancy of the skin cancer compared to a low expression level.

[0019] (13) According to another aspect of the present invention, there is provided a biomarker for predicting recurrence of skin cancer, the biomarker comprising glucose-6-phosphate dehydrogenase in a sample collected from a patient suffering from the skin cancer, wherein a high expression level of the biomarker in the sample indicates a higher possibility of recurrence of the skin cancer than a low expression level.

[0020] (14) According to another aspect of the present invention, there is provided a method for predicting the likelihood of an immune-related adverse event of Grade 3 or higher caused by administration of an immune checkpoint inhibitor to a skin cancer patient. The method of this aspect includes a step of evaluating, when the effectiveness of the immune checkpoint inhibitor is evaluated to be high, that there is a high likelihood of an immune-related adverse event of Grade 3 or higher caused by administration of the immune checkpoint inhibitor. According to the method of this aspect, the likelihood of an immune-related adverse event of Grade 3 or higher caused by administration of an immune checkpoint inhibitor can be predicted using the expression level of glucose-6-phosphate dehydrogenase in a sample collected from a skin cancer patient as an index.

[0021] (15) According to another aspect of the present invention, there is provided a biomarker for predicting the likelihood of onset of an immune-related adverse event of Grade 3 or higher caused by administration of an immune checkpoint inhibitor for skin cancer. The biomarker of this aspect includes glucose-6-phosphate dehydrogenase in a sample collected from a patient with skin cancer, and indicates that a low expression level of the biomarker in the sample is more likely to result in an immune-related adverse event of Grade 3 or higher caused by administration of the immune checkpoint inhibitor than a high expression level.

[0022] (16) According to another embodiment of the present invention, a measuring kit is provided. The measuring kit of this embodiment is used for at least one of predicting the prognosis of skin cancer, evaluating the effectiveness of an immune checkpoint inhibitor against the skin cancer, evaluating the malignancy of the skin cancer, predicting the recurrence of the skin cancer, predicting the possibility of an immune-related adverse event of Grade 3 or higher caused by the administration of the immune checkpoint inhibitor, and evaluating the immune activity of the skin cancer, and includes a substance capable of detecting a correlation amount that correlates with the expression level of glucose-6-phosphate dehydrogenase in a sample collected from a patient with the skin cancer. According to the measuring kit of this embodiment, a correlation amount that correlates with the expression level of glucose-6-phosphate dehydrogenase in a sample collected from a patient with skin cancer can be easily detected.

[0023] (17) In the above-described measuring kit, the substance capable of detecting the correlation amount may comprise at least one selected from the group consisting of a substance capable of binding to the glucose-6-phosphate dehydrogenase or a fragment thereof, a substance capable of binding to a gene encoding the glucose-6-phosphate dehydrogenase, and glucose-6-phosphate. [Brief description of the drawings]

[0024] [Figure 1] FIG. 1 shows Kaplan-Meier curves of G6PD mRNA expression levels. [Diagram 2] FIG. 1 is an explanatory diagram showing the results of G6PD expression analysis by immunohistochemical staining. [Diagram 3] FIG. 1 shows Kaplan-Meier curves of G6PD expression levels. [Figure 4] A graph showing the Kaplan-Meier curve of PD-L1 expression levels. [Diagram 5] A graph showing the results of correlation analysis of PD-L1 expression levels. [Figure 6] FIG. 1 is an explanatory diagram showing the time course of G6PD activity in Case 1. [Figure 7] FIG. 1 is an explanatory diagram showing the time course of G6PD activity in Case 2. [Figure 8] FIG. 1 is an explanatory diagram showing the transition of G6PD activity in Case 4. [Figure 9] FIG. 1 is an explanatory diagram showing the results of G6PD expression analysis by immunohistochemical staining in malignant melanoma. [Figure 10] FIG. 1 shows the Kaplan-Meier curve of G6PD expression levels in malignant melanoma. [Figure 11] FIG. 1 is an explanatory diagram showing the relationship between the presence or absence of immune-related adverse events of Grade 3 or higher in malignant melanoma and the expression level of G6PD. [Figure 12] FIG. 1 is an explanatory diagram showing the results of G6PD expression analysis by immunohistochemical staining in cutaneous angiosarcoma. [Figure 13] FIG. 1 is an explanatory diagram showing the results of G6PD expression analysis by next-generation sequencing in cutaneous angiosarcoma. [Figure 14]FIG. 1 is an explanatory diagram showing the results of GSEA analysis of cutaneous angiosarcoma with high G6PD expression. [Figure 15] FIG. 1 is an explanatory diagram showing the results of GSEA analysis of cutaneous angiosarcoma with high G6PD expression. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0025] According to one embodiment of the present disclosure, a method for predicting the prognosis of skin cancer is provided. The types of skin cancer include, but are not limited to, Merkel cell carcinoma, malignant melanoma, squamous cell carcinoma, extramammary Paget's disease, cutaneous angiosarcoma, and the like. The skin cancer patient may be a patient with multiple skin cancers, and may be a patient before or after treatment. More specifically, the skin cancer patient may be a patient before or after surgery, a patient before or after immunotherapy, a patient before or after chemotherapy, or a patient before or after radiation therapy. The sex and age of the skin cancer patient are also not particularly limited. The animal species of the patient in this embodiment is mainly mammals. The mammals include, but are not particularly limited to, primates such as humans and chimpanzees, dogs, cats, rabbits, and the like.

[0026] As shown in the Examples below, the present inventors have found that high levels of glucose-6-phosphate dehydrogenase expression in skin cancers tend to result in a worse prognosis than low levels of this enzyme expression.

[0027] The method for predicting the prognosis of skin cancer in this embodiment includes: (I) determining a correlation amount that correlates with the expression level of glucose-6-phosphate dehydrogenase in a sample collected from a skin cancer patient (hereinafter also referred to as step (I)); and (II) determining that when the correlation amount is high, the prognosis of skin cancer is worse than when the correlation amount is low (hereinafter also referred to as step (II)).

[0028] The sample used in step (I) may be a tissue containing a carcinoma isolated from a skin cancer patient, blood or serum collected from a skin cancer patient, etc. The tissue may be, but is not limited to, a formalin-fixed paraffin-embedded (FFPE) specimen, at least a portion of a tumor excised by surgery or the like, a sample containing a plurality of tissue-constituting cells collected by biopsy or the like, etc.

[0029] In this embodiment, glucose-6-phosphate dehydrogenase refers to an enzyme with enzyme number EC1.1.1.49. Glucose-6-phosphate dehydrogenase (hereinafter also referred to as "G6PD") catalyzes a reaction in which a substrate, glucose-6-phosphate, is oxidized in the presence of a coenzyme such as nicotinamide adenine dinucleotide (NAD) or nicotinamide adenine dinucleotide phosphate (NADP), to produce glucono-1,5-lactone-6-phosphate and reduced NAD (NADH) or reduced NADP (NADPH), or the reverse reaction.

[0030] In this embodiment, the correlation amount correlated with the expression amount of G6PD means a correlation amount that is positively correlated with the expression amount of G6PD. Such a correlation amount is not particularly limited, but may be, for example, the expression amount of G6PD itself, the expression amount of mRNA encoding G6PD, or the activity of G6PD.

[0031] The expression level of G6PD can be determined, for example, by Western blotting, dot blotting, immunoprecipitation, ELISA, immunohistochemical staining, etc. based on antigen-antibody reaction using anti-G6PD antibody (Anti-G6PD-antibody) or a fragment thereof. The expression level of mRNA encoding G6PD can be determined, for example, by next-generation sequencing analysis, Northern hybridization, dot hybridization, RT-PCR, real-time PCR, etc. The activity of G6PD can be measured, for example, using a kit for measuring G6PD activity using glucose-6-phosphate as a substrate. The expression level of G6PD determined by immunostaining has little variation within a case and is excellent as a biomarker. In addition, the expression level of G6PD determined by a serum test using serum is suitable for monitoring because of its high sensitivity.

[0032] In step (II), for example, (i) when the expression level of G6PD or the expression level of mRNA encoding G6PD is high, the prognosis of skin cancer may be judged to be poorer than when the expression level is low, or (ii) when the activity of G6PD is high, the prognosis of skin cancer may be judged to be poorer than when the activity of G6PD is low. The magnitude of the expression level or activity may be compared with a predetermined reference value, or may be a comparison of values ​​in samples taken at different times from the same patient. In this embodiment, "poor prognosis" means that the malignancy of skin cancer is high and the condition tends to worsen. In other words, when the prognosis is poor, there is a possibility of metastasis or recurrence.

[0033] For example, when the expression level of G6PD is high in the resection specimen as a sample, it is considered that the skin cancer is a high-risk group with high malignancy, and postoperative radiation therapy or the like may be added. Also, for example, when the expression level of G6PD is low in the resection specimen as a sample, it is considered that the skin cancer is a low-risk group with low malignancy, and the progress may be observed without adding postoperative radiation therapy or the like. In this way, the skin cancer prognosis prediction method in this embodiment can be used as a method for assisting in the decision of a treatment policy for skin cancer. In other words, the skin cancer prognosis prediction method can be said to be a method for predicting the prognosis of skin cancer.

[0034] According to another embodiment of the present disclosure, a method for evaluating the effectiveness of an immune checkpoint inhibitor against skin cancer is provided. In this embodiment, the immune checkpoint inhibitor refers to a drug that inhibits signal transduction by an immune checkpoint by inhibiting the binding between the immune checkpoint and its ligand. Examples of immune checkpoint inhibitors include anti-PD-L1 antibody drugs and anti-PD-1 antibody drugs. The anti-PD-L1 antibody drug is composed of a human monoclonal antibody, and by binding to PD-L1 (programmed cell death ligand 1) expressed in tumor cells, it inhibits the binding of PD-L1 to PD-1 (programmed cell death 1) expressed in T cells, thereby maintaining the activation of T cells. The anti-PD-1 antibody drug inhibits the binding of PD-L1 to PD-1 by binding to PD-1, thereby maintaining the activation of T cells.

[0035] As shown in the Examples below, the inventors of the present application have found that the expression level of G6PD in skin cancer is inversely correlated with PD-L1, and the higher the expression level of G6PD, the lower the expression level of PD-L1. It can be said that the higher the expression level of G6PD, i.e., the lower the estimated expression level of PD-L1, the more likely it is that the tumor has low immunological activity.

[0036] In this embodiment, the method for evaluating the effectiveness of an immune checkpoint inhibitor against skin cancer includes the above-mentioned step (I), i.e., a step of determining a correlation amount that correlates with the expression level of glucose-6-phosphate dehydrogenase in a sample collected from a patient, and (III) a step (hereinafter also referred to as step (III)) of evaluating that the effectiveness of the immune checkpoint inhibitor is higher when the correlation amount is low than when the correlation amount is high.

[0037] In step (III), for example, (i) when the expression level of G6PD or the expression level of mRNA encoding G6PD is low, the effectiveness of the immune checkpoint inhibitor may be determined to be higher than when the expression level is high, and (ii) when the activity of G6PD is low, the effectiveness of the immune checkpoint inhibitor may be evaluated to be higher than when the activity of G6PD is high. The magnitude of the expression level or activity may be compared with a predetermined reference value, or the values ​​of samples taken at different times from the same patient may be compared.

[0038] Considering the mechanism of immune checkpoint inhibitors such as anti-PD-L1 antibodies, it is estimated that when the expression level of PD-L1 is low, the effect of immune checkpoint inhibitors such as anti-PD-L1 antibodies is low, and when the expression level of PD-L1 is high, the effect of immune checkpoint inhibitors such as anti-PD-L1 antibodies is high. Therefore, in the above step (III), when the expression level of G6PD is low, i.e., when the expression level of PD-L1 is estimated to be high due to the correlation between G6PD and PD-L1, the efficacy of immune checkpoint inhibitors such as anti-PD-L1 antibodies is evaluated to be high compared to when the expression level of G6PD is high, i.e., when the expression level of PD-L1 is estimated to be low due to the correlation between G6PD and PD-L1.

[0039] For example, when the expression level of G6PD in a sample is low and the expression level of PD-L1 is low, it is expected that the expression level of PD-L1 is temporarily low due to the heterogeneity of PD-L1. Even in such a case, the efficacy of an immune checkpoint inhibitor such as an anti-PD-L1 antibody drug may be evaluated as high because the expression level of G6PD is low. Since G6PD has a small numerical variation within the same case, it is superior as an evaluation index for the efficacy of an immune checkpoint inhibitor, etc., compared with PD-L1, which has a large numerical variation within the same case. The evaluation method for evaluating the efficacy of an immune checkpoint inhibitor against skin cancer in this embodiment can be used as a method for assisting in the determination of a treatment policy for skin cancer, such as the determination of the administration of an immune checkpoint inhibitor.

[0040] According to another embodiment of the present disclosure, there is provided a method for determining whether or not to administer an immune checkpoint inhibitor to a patient with skin cancer. This method includes a step (IV) of determining to administer or continue the administration of an immune checkpoint inhibitor to the patient (hereinafter also referred to as step (IV)) when the effectiveness of the immune checkpoint inhibitor against skin cancer is evaluated to be high in step (III) of the above-mentioned evaluation method for evaluating the effectiveness of the immune checkpoint inhibitor against skin cancer.

[0041] In step (IV), it may be determined that an immune checkpoint inhibitor will be newly administered to a patient who has not yet been administered an immune checkpoint inhibitor, or that the administration of an immune checkpoint inhibitor will be continued to a patient who is already being administered an immune checkpoint inhibitor.

[0042] For example, when the expression level of G6PD in the sample is high and the expression level of PD-L1 is low, the effect of immune checkpoint inhibitors such as anti-PD-L1 antibody drugs is evaluated to be low, so instead of administering immune checkpoint inhibitors, surgical therapy such as lymphadenectomy may be selected. In addition, for example, when the blood G6PD level is maintained at a low level by administration of immune checkpoint inhibitors such as anti-PD-L1 antibody drugs, the effectiveness of such immune checkpoint inhibitors is evaluated to be high, so administration of such immune checkpoint inhibitors may be continued. In addition, for example, when the blood G6PD level tends to increase after administration of immune checkpoint inhibitors, the effectiveness of such immune checkpoint inhibitors is evaluated to be low, so additional treatment such as radiation therapy may be considered. In addition, for example, when the expression level of G6PD in the sample is low and the estimated expression level of PD-L1 is high, it is considered to be a tumor with high immune activity, so immune checkpoint inhibitors may be introduced while paying close attention to side effects.

[0043] As described above, the inventors of the present application have found that the higher the expression level of G6PD, i.e., the lower the estimated expression level of PD-L1, the more likely the tumor has low immunological activity.Therefore, the expression level of G6PD can be used as an index to evaluate the immune activity of skin cancer.That is, as another embodiment of the present disclosure, a method for evaluating the immune activity of skin cancer is provided.This evaluation method includes a step of determining a correlation amount that correlates with the expression level of G6PD in a sample collected from a skin cancer patient, and a step of determining that the immune activity of skin cancer is high when the correlation amount is high compared to when the correlation amount is low.

[0044] According to another embodiment of the present disclosure, a biomarker is provided. The biomarker includes G6PD in a sample collected from a patient with skin cancer. The biomarker can be used to predict the prognosis of skin cancer, to evaluate the effectiveness of an immune checkpoint inhibitor against skin cancer, to evaluate the malignancy of skin cancer, and to predict the recurrence of skin cancer. The biomarker can also be used to predict the likelihood of an immune-related adverse event of Grade 3 or higher caused by the administration of an immune checkpoint inhibitor against skin cancer, and to evaluate the immune activity of skin cancer. According to the present disclosure, the use of G6PD as a biomarker in a sample collected from a patient with skin cancer is envisioned. More specifically, the use of G6PD as a biomarker in a sample collected from a patient with skin cancer is envisioned for at least one of the following: prediction of the prognosis of skin cancer, evaluation of the effectiveness of an immune checkpoint inhibitor against skin cancer, evaluation of the malignancy of skin cancer, prediction of the recurrence of skin cancer, prediction of the likelihood of an immune-related adverse event of Grade 3 or higher caused by the administration of an immune checkpoint inhibitor, and evaluation of the immune activity of skin cancer.

[0045] A biomarker for predicting the prognosis of skin cancer indicates that a high expression level of the biomarker indicates a poorer prognosis of skin cancer compared to a low expression level. A biomarker for evaluating the efficacy of an immune checkpoint inhibitor against skin cancer indicates that a high expression level of the biomarker indicates a higher efficacy of the immune checkpoint inhibitor compared to a low expression level. A biomarker for evaluating the malignancy of skin cancer indicates that a high expression level of the biomarker indicates a higher malignancy of skin cancer compared to a low expression level.

[0046] A biomarker for predicting recurrence of skin cancer indicates that the possibility of recurrence of skin cancer is higher when the expression level of the biomarker is high in a sample collected from a patient suffering from skin cancer than when the expression level is low. For example, when the blood G6PD level increases during postoperative follow-up, the possibility of recurrence of skin cancer is expected, and therefore an imaging test may be performed.

[0047] A biomarker for assessing immune activity in skin cancer indicates high immune activity in skin cancer when the expression level of the biomarker is high in a sample taken from a patient suffering from skin cancer, compared to when the expression level is low.

[0048] A biomarker for predicting the likelihood of developing Grade 3 or higher immune-related adverse events (irAEs) due to the administration of immune checkpoint inhibitors to skin cancer shows that when the expression level of the biomarker is low in a sample collected from a skin cancer patient, the likelihood of developing Grade 3 or higher irAEs due to the administration of immune checkpoint inhibitors is higher than when the expression level is high. This is supported by the fact that, as shown in the examples below, when the expression level of G6PD is low, the frequency of developing severe irAEs due to the administration of immune checkpoint inhibitors is higher than when the expression level of G6PD is high.

[0049] According to another embodiment of the present disclosure, a method for predicting the likelihood of onset of Grade 3 or higher immune-related adverse events (irAEs) caused by administration of an immune checkpoint inhibitor to a patient with skin cancer is provided. This method includes a step (V) of evaluating that there is a high likelihood of onset of Grade 3 or higher irAEs caused by administration of an immune checkpoint inhibitor when the efficacy of the immune checkpoint inhibitor is evaluated to be high in step (III) of the above-mentioned evaluation method for evaluating the efficacy of the immune checkpoint inhibitor against skin cancer (hereinafter also referred to as step (V)). In addition, the method for predicting the likelihood of onset of Grade 3 or higher irAEs caused by administration of an immune checkpoint inhibitor to a patient with skin cancer may include the above-mentioned step (I), that is, (I) a step of determining a correlation amount that correlates with the expression amount of G6PD in a sample collected from the patient, and (VI) a step (hereinafter also referred to as step (VI)) of evaluating that there is a higher likelihood of onset of Grade 3 or higher irAEs caused by administration of an immune checkpoint inhibitor when the correlation amount is low, compared to when the correlation amount is high.

[0050] For example, when the expression level of G6PD in a sample is low and the expression level of estimated PD-L1 is high, it is considered to be a tumor with high immune activity, so that the effectiveness of immune checkpoint inhibitors is likely to be high. In such tumors with high immune activity, the administration of immune checkpoint inhibitors is likely to cause Grade 3 or higher irAE, so it is expected that irAE should be carefully monitored when administering immune checkpoint inhibitors. Thus, the method for predicting the possibility of Grade 3 or higher irAE caused by the administration of immune checkpoint inhibitors to skin cancer patients in this embodiment can be used as a method for assisting in the determination of a treatment policy for skin cancer, such as the decision on the administration of immune checkpoint inhibitors.

[0051] According to another embodiment of the present disclosure, a measurement kit is provided. The measurement kit is used for at least one of predicting the prognosis of skin cancer, evaluating the effectiveness of an immune checkpoint inhibitor against skin cancer, evaluating the malignancy of skin cancer, predicting the recurrence of skin cancer, predicting the likelihood of an immune-related adverse event of Grade 3 or higher caused by the administration of an immune checkpoint inhibitor, and evaluating the immune activity of skin cancer, and includes a substance capable of detecting a correlation amount that correlates with the expression level of G6PD in a sample collected from a patient with skin cancer. In other words, the measurement kit includes a companion diagnostic agent that can be used for companion diagnosis of skin cancer. The substance capable of detecting a correlation amount that correlates with the expression level of G6PD may include at least one selected from the group consisting of a substance capable of binding to G6PD or a fragment of G6PD, a substance capable of binding to a gene encoding G6PD, and glucose-6-phosphate.

[0052] Examples of substances capable of binding to G6PD or a fragment of G6PD include anti-G6PD antibodies and fragments thereof. Examples of substances capable of binding to a gene encoding G6PD include a primer set capable of amplifying the G6PD gene and a probe that specifically hybridizes to the G6PD gene. Glucose-6-phosphate may be included in the measurement kit as a substrate for measuring the activity of G6PD.

[0053] The form of the measurement kit is not particularly limited, and may be, for example, in a dry state, or in a state in which the above-mentioned substance capable of detecting the correlation amount correlated with the expression amount of G6PD is dissolved in a solution. The above-mentioned substance may be labeled with a labeling substance such as a secondary antibody, a fluorescent substance, or a radioisotope, and may be immobilized on a support such as a microarray substrate, a microtiter plate, or resin or metal beads. In addition to the above-mentioned substance, the measurement kit may contain various components such as reverse transcriptase, DNA polymerase, dNTP, oligo dT primer, random primer, RNase inhibitor, RNase H, a labeling substance, and a buffer solution. Furthermore, the measurement kit may contain various devices that can be used to determine the expression amount of G6PD, instructions for use, etc. EXAMPLES

[0054] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to the following examples.

[0055] <Merkel cell carcinoma> 1. Sample Formalin-fixed paraffin-embedded (FFPE) specimens and serum from cases of Merkel cell carcinoma in which histopathological examinations had been performed and postoperative progress over a certain period of time had been made clear were used as samples. There were 90 FFPE specimens from 71 cases, and 50 serum specimens from 19 cases. The ages of the patients in the FFPE specimen cases ranged from 40 to 98 years, with an average age of 77.27 years, and the male-female ratio was 26:45. The tumor sites in these cases were head and neck in 66.2% (face in 42 cases, neck in 2 cases, ear in 1 case, head in 2 cases), limbs in 29.6% (upper arm in 4 cases, forearm in 3 cases, finger in 3 cases, thigh in 6 cases, lower leg in 3 cases, foot in 2 cases), and trunk in 2.8% (buttocks in 2 cases). Of these, there were 6 cases of spontaneous regression.

[0056] 2. Search for biomarkers that correlate with PD-L1 2-1.RNA extraction From unstained slides prepared from FFPE specimens, the tumor area and the surrounding inflammatory cell infiltrate area were macrodissected using an 18G needle, and RNA was extracted using the AllPrep DNA / RNA FFPE Kit (Qiagen). The RIN value and DV200 (median value of RNA fragment size) were measured using a bioanalyzer (Agilent2100, Agilent), and 44 samples with DV200 of 30% or more were selected.

[0057] 2-2. Next-generation sequencing analysis Next-generation sequencing (NGS) analysis was performed on 44 samples using the Immune Response Panel (Ampliseq) on a MiniSeq (Illumina) system. The obtained data was analyzed on BaseSpace (registered trademark) Sequence Hub (Illumina).

[0058] When comparing the high and low PD-L1 expression groups, the only factor correlated with PD-L1 was glucose-6-phosphate dehydrogenase (G6PD), with a q value (p-adjusted) of 0.05 or less. Low PD-L1 levels were associated with significantly higher G6PD levels (p=0.00011, q=0.040). In other words, G6PD was found to show an inverse correlation with PD-L1.

[0059] 3. Examination of the correlation between G6PD mRNA expression level and prognosis We investigated whether there was a correlation between the expression level of G6PD mRNA and the presence or absence of metastasis in patients with skin cancer. As a result, the expression level of G6PD mRNA was correlated with the presence or absence of metastasis in patients with skin cancer. Cases with high levels of G6PD mRNA expression were significantly more likely to develop lymph node metastasis and distant metastasis during the course of the disease (p=0.00016, q=0.018). A receiver operating characteristic curve was drawn for the presence or absence of metastasis, and a G6PD mRNA expression level of high was calculated as 1071 CPM (counts per million) or higher, and a G6PD mRNA expression level of low was calculated as 1071 CPM or lower.

[0060] FIG. 1 is a diagram showing the Kaplan-Meier curve of the mRNA expression level of G6PD. In FIG. 1, the Kaplan-Meier curve of the survival rate of each group is shown for a group with high G6PD mRNA expression level and a group with low G6PD mRNA expression level. In FIG. 1, the group with high G6PD mRNA expression level is shown by a solid line, and the group with low G6PD mRNA expression level is shown by a dashed line. In FIG. 1, the vertical axis indicates the overall survival rate, the horizontal axis indicates the number of years, and the P value indicates the log-rank test value. As shown in FIG. 1, when the mRNA expression level of G6PD is high, there was a significant correlation between the survival rate and the prognosis, that is, when the mRNA expression level of G6PD is low, compared with when the mRNA expression level of G6PD is low (P=0.036).

[0061] 4.Immunohistological staining 4-1. Immunohistochemical staining of G6PD Immunohistochemical staining of G6PD was performed using slides prepared from FFPE specimens. Anti-G6PD antibody (HPA000247, Sigma-Ardrich) was used. Images were taken and analyzed using an all-in-one fluorescence microscope BZ-X800 (Keyence). The staining results were digitized, and the number of positive cells and expression intensity were quantified for evaluation.

[0062] Figure 2 is an explanatory diagram showing the results of G6PD expression analysis by immunohistochemical staining. In Figure 2, an example of an image of immunohistochemical staining in a group showing high G6PD expression is shown on the left side of the page, and an example of an image of immunohistochemical staining in a group showing low G6PD expression is shown on the right side of the page. The expression level of G6PD in immunohistochemical staining was evaluated by visual observation of the stained specimen under a microscope, with a staining rate of 50% or more being rated as high and less than 50% being rated as low.

[0063] FIG. 3 is a diagram showing the Kaplan-Meier curve of the expression level of G6PD. In FIG. 3, the Kaplan-Meier curve of the survival rate of each group is shown for a group with high expression level of G6PD and a group with low expression level of G6PD. In FIG. 3, the group with high expression level of G6PD is shown by a dashed line, and the group with low expression level of G6PD is shown by a solid line. In FIG. 3, the vertical axis shows the overall survival rate, and the horizontal axis shows the overall survival period (OS: Overall Survival). As shown in FIG. 3, a significant correlation was observed between a case with high expression level of G6PD and a case with low expression level of G6PD, i.e., a poor prognosis (P=0.036).

[0064] 4-2. Immunohistochemical staining of PD-L1 in primary tumors As a comparative example, PD-L1 immunohistochemical staining was performed on slides prepared from FFPE specimens of primary tumors. The antibody used was anti-PD-L1 antibody (28-8, ab205921, Abcam). Images were taken and analyzed using an all-in-one fluorescence microscope BZ-X800 (Keyence). The staining results were digitized, and the number of positive cells and expression intensity were quantified for evaluation.

[0065] FIG. 4 is a diagram showing the Kaplan-Meier curve of the expression level of PD-L1. FIG. 5 is a diagram showing the results of correlation analysis of the expression level of PD-L1. FIG. 4 shows Kaplan-Meier curves of the survival rates of the groups with high expression level of PD-L1 and the groups with low expression level of PD-L1. In FIG. 4, the group with high expression level of PD-L1 is shown by a solid line, and the group with low expression level of G6PD is shown by a dashed line. In FIG. 4, the vertical axis shows the overall survival rate, and the horizontal axis shows the number of years. In FIG. 5, the vertical axis shows the expression level (pixel value) of PD-L1, and the horizontal axis shows the number of months. As shown in FIG. 4 and FIG. 5, no correlation with survival rate was observed in the comparison between the group with high expression of PD-L1 and the group with low expression of PD-L1 (r=0.068, CI [-0.19 to 0.31], P=0.59).

[0066] 4-3. Considerations of immunohistochemical staining The results of immunohistochemical staining for G6PD and PD-L1 showed no correlation between PD-L1 expression and prognosis, whereas there was a correlation between G6PD expression and prognosis, indicating that G6PD can be used as a better prognostic marker than PD-L1.

[0067] 5.G6PD activity 5-1. Measurement of G6PD activity in serum G6PD activity was measured in 50 patient serum samples from 19 cases using a G6PD assay kit (Abcam, ab102529). The average activity in the 50 samples was 11.45 mU / ml.

[0068] 5-2. Relationship between G6PD activity and progression Of the 50 specimens, the progress of Merkel cell carcinoma was observed for three specimens (Cases 1-3) in which G6PD activity was particularly high (19mU / ml or higher). In addition, the progress of Merkel cell carcinoma was observed for a case (Case 4) in which G6PD activity had decreased. In Figures 6-8 shown below, the vertical axis indicates G6PD activity (mU / ml) and the horizontal axis indicates the period.

[0069] (1) Case 1 FIG. 6 is an explanatory diagram showing the transition of G6PD activity in case 1. Case 1 is a stage IV case, and the location of the primary lesion was the right mandible. In case 1, surgical resection and radiation therapy were performed, but metastasis was observed in lymph nodes throughout the body about 9 months after surgical resection. After that, combination therapy with CBDGB and VP-16 was performed, but since no therapeutic effect was observed, a clinical trial of avelumab, an immune checkpoint inhibitor, was performed, but the patient died of the current disease several months later. High G6PD activity was observed at the time of death, and the G6PD activity was 21.84 mU / ml.

[0070] (2) Case 2 Figure 7 is an explanatory diagram showing the transition of G6PD activity in case 2. Case 2 was a stage IV case, and the primary lesion was located in the left lower leg. In case 2, surgical resection, SLNB, and radiation therapy were performed, but local recurrence was observed about one year after the resection and skin graft. High G6PD activity was observed at the time of local recurrence, and the G6PD activity was 19.63 mU / ml.

[0071] (3) Case 3 Case 3 was a stage II case with the primary tumor located in the anterior chest. Surgical resection and radiation therapy were performed, but the patient died of other diseases about 2 months after the completion of radiation therapy. High G6PD activity was observed before the patient died of other diseases, and the G6PD activity was 19.37 mU / ml.

[0072] (4) Case 4 FIG. 8 is an explanatory diagram showing the progress of G6PD activity in case 4. Case 4 is a stage III case, and the primary lesion was located at the base of the nose. In case 4, after surgical resection, PET / CT showed accumulation in the right submandibular lymph node, so administration of avelumab, an immune checkpoint inhibitor, was started. PET / CT showed increased accumulation three months later, so IMRT (intensity modulated radiation therapy) was administered in combination, resulting in CR (complete remission). G6PD activity was significantly reduced by the combination of avelumab and IMRT.

[0073] 6. Summary From the above results, it was found that in Merkel cell carcinoma as skin cancer, the expression level of G6PD was inversely correlated with PD-L1, and the higher the expression level of G6PD, the lower the expression level of PD-L1. In addition, a correlation was observed between a high expression level of G6PD and a poorer prognosis compared to a low expression level of G6PD. In addition, it was suggested that a high activity of G6PD in serum and a high expression level of G6PD are more likely to cause recurrence of skin cancer than a low activity of G6PD and a low expression level of G6PD. In other words, it was suggested that a high expression level of G6PD is associated with a higher malignancy of skin cancer and a worsening of the condition compared to a low expression level of G6PD. Therefore, it was found that G6PD is effective as a biomarker for predicting the prognosis of skin cancer, a biomarker for evaluating the malignancy of skin cancer, and a biomarker for predicting the recurrence of skin cancer. Therefore, it was found that by using the expression level of G6PD as an index, it is possible to predict the prognosis of skin cancer, evaluate the effectiveness of immune checkpoint inhibitors, determine whether or not to administer immune checkpoint inhibitors, and evaluate the malignancy of skin cancer. In addition, since the expression level of G6PD was higher, the expression level of PD-L1 was lower, suggesting that the expression level of G6PD can be used as an index to evaluate the immune activity of skin cancer.

[0074] <Malignant melanoma> 1. Sample Samples from primary lesions in 30 cases of malignant melanoma that had undergone immune checkpoint therapy were used. Immune checkpoint therapy means that immune checkpoint inhibitors were administered. The RNA extraction method, next-generation sequencing analysis method, immunohistochemical staining method for G6PD, and evaluation criteria for G6PD expression level were the same as those described in the example of Merkel cell carcinoma.

[0075] 2.Immunohistological staining FIG. 9 is an explanatory diagram showing the results of expression analysis of G6PD by immunohistochemical staining in malignant melanoma. In FIG. 9, an example of an image of immunohistochemical staining in a group showing high expression of G6PD is shown on the left side of the page, and an example of an image of immunohistochemical staining in a group showing low expression of G6PD is shown on the right side of the page. As a result of immunohistochemical staining of G6PD for 30 samples, 12 cases showed high expression of G6PD and 18 cases showed low expression of G6PD. The expression level of G6PD in immunohistochemical staining was evaluated by visual observation of the stained specimen under a microscope, and a staining rate of 50% or more was determined to be high (high expression), and less than 50% was determined to be low (low expression).

[0076] 3. Correlation between G6PD mRNA expression level and prognosis Survival curves were created using the Kaplan-Meier method for samples from 17 cases in which no immune-related adverse events (irAEs) of Grade 3 or higher were observed after administration of immune checkpoint inhibitors.

[0077] FIG. 10 is a diagram showing the Kaplan-Meier curve of G6PD expression in malignant melanoma. In FIG. 10, the Kaplan-Meier curve of the survival rate of each group is shown for a group with high G6PD expression and a group with low G6PD expression. In FIG. 10, the group with high G6PD expression is shown by a thin line, and the group with low G6PD expression is shown by a thick line. In FIG. 10, the vertical axis shows the progression-free survival rate, and the horizontal axis shows the number of years after the start of administration of immune checkpoint inhibitors (Years after ICI start). From the results of FIG. 10, it was found that the group with low G6PD expression has a higher survival rate, i.e., a better prognosis, compared to the group with high G6PD expression. From this, it can be said that the same tendency was observed when the type of skin cancer was malignant melanoma as when the type of skin cancer was Merkel cell carcinoma. Therefore, as in the case of Merkel cell carcinoma, it was suggested that in malignant melanoma, the expression level of G6PD can be used as an indicator to predict the prognosis of skin cancer, evaluate the effectiveness of immune checkpoint inhibitors, determine whether or not to administer immune checkpoint inhibitors, evaluate the malignancy of skin cancer, predict the possibility of skin cancer recurrence, and evaluate the immune activity of skin cancer.

[0078] 4. Relationship between G6PD expression levels and immune-related adverse events The relationship between G6PD expression levels and immune-related adverse events was analyzed. Whether or not immune-related adverse events (irAEs) of Grade 3 or higher were observed following administration of immune checkpoint inhibitors was analyzed in the high G6PD expression group and the low G6PD expression group.

[0079] FIG. 11 is an explanatory diagram showing the relationship between the presence or absence of immune-related adverse events of Grade 3 or higher in malignant melanoma and the expression level of G6PD. The figure on the right side of the page of FIG. 11 is a 2×2 contingency table of Fisher's test regarding the expression level of G6PD and the presence or absence of irAEs of Grade 3 or higher, and the figure on the left side of the page of FIG. 11 is a graph created based on the 2×2 contingency table. In FIG. 11, "irAE+" indicates that irAEs of Grade 3 or higher were observed, and "irAE-" indicates that irAEs of Grade 3 or higher were not observed. From the results shown in FIG. 11, it was found that the number of cases in which irAEs of Grade 3 or higher were observed was significantly higher in the group with low expression of G6PD (p=0.019). In other words, a significant correlation was found between the case where the expression level of G6PD is low and the case where the expression level of G6PD is high, and the possibility of developing irAEs of Grade 3 or higher due to administration of immune checkpoint inhibitors. Therefore, it was found that the expression level of G6PD can be used as an indicator to predict the likelihood of developing Grade 3 or higher irAEs caused by administration of immune checkpoint inhibitors to skin cancer patients.

[0080] <Cutaneous angiosarcoma> 1. Sample Fourteen specimens from seven cases of cutaneous angiosarcoma were used as samples. The RNA extraction method, next-generation sequencing analysis method, G6PD immunohistochemical staining method, and evaluation criteria for G6PD expression level were the same as those described in the examples for Merkel cell carcinoma.

[0081] (1) Case A Case A had a primary tumor in the head, and was treated with wPTX and radiation therapy. Although wPTX was administered, the patient's performance status (PS) worsened, and the patient was placed on best supportive care (BSC). Later, lung metastasis was observed, leading to death from the primary disease. The overall survival (OS) for Case A was 389 days.

[0082] (2) Case B In case B, the primary tumor was located in the head, and wPTX therapy, radiation therapy, and eribulin therapy were administered as treatment. In case B, parotid lymph node metastasis was observed during wPTX administration, so the administration was changed to eribulin therapy, but the patient developed pneumothorax (PD). The OS in case B was 488 days.

[0083] (3) Case C In case C, the primary tumor was located in the head, and the treatment consisted of surgery, immunotherapy using IL-2, wDTX therapy, wPTX therapy, and radiation therapy. In case C, pneumothorax occurred due to wDTX administration, so the treatment was changed to wPTX. The administration was discontinued at the patient's request, and chemotherapy initiation is currently being considered. The OS in case C is over 4,826 days.

[0084] 2.Immunohistological staining FIG. 12 is an explanatory diagram showing the results of G6PD expression analysis by immunohistochemical staining in cutaneous angiosarcoma. In FIG. 12, the images of immunohistochemical staining in cutaneous angiosarcoma cases A, B, and C are shown at the top of the page, and their enlarged images are shown at the bottom of the page. In cases A and B, the results are shown using biopsy specimens as samples, and in case C, the results are shown using surgical specimens as samples. From the results shown in FIG. 12, it was confirmed that G6PD was highly expressed in cases A and B, and that G6PD was lowly expressed in case C. In addition, the relationship between the expression level of G6PD and prognosis suggested that a low expression level of G6PD indicates a good prognosis, and a high expression level of G6PD tends to indicate a poor prognosis. From this, it can be said that the same tendency was observed when the type of skin cancer was cutaneous angiosarcoma as when the type of skin cancer was Merkel cell carcinoma. Therefore, in cutaneous angiosarcoma, as in Merkel cell carcinoma, it is suggested that the expression level of G6PD can be used as an indicator to predict the prognosis of skin cancer, evaluate the effectiveness of immune checkpoint inhibitors, determine whether or not to administer immune checkpoint inhibitors, evaluate the malignancy of skin cancer, predict the possibility of skin cancer recurrence, and evaluate the immune activity of skin cancer.

[0085] 3. Next-generation Sequencing Analysis 13 is an explanatory diagram showing the results of G6PD expression analysis by next-generation sequencing analysis of cutaneous angiosarcoma. Of the 14 specimens used as samples, 10 were G6PD positive and 4 were G6PD negative. Here, G6PD positivity indicates that the staining rate of tumor cells in the results of immunohistochemical staining is 50% or more, indicating high G6PD expression. G6PD negativity indicates that the staining rate of tumor cells in the results of immunohistochemical staining is less than 50%, indicating low G6PD expression.

[0086] 4.GSEA Analysis Gene set enrichment analysis (GSEA analysis) was performed on genes that were highly expressed in the G6PD high expression group using the GSEA software (https: / / www.gsea-msigdb.org / gsea / ). The c5 Gene Ontology (GO) gene set collection provided by MSigDB (Molecular Signatures Database) was used as the gene set.

[0087] Figures 14 and 15 are explanatory diagrams showing the results of GSEA analysis in the group with high G6PD expression in cutaneous angiosarcoma. The results shown in Figures 14 and 15 indicate that the group with high G6PD RNA expression has abnormal antibody production. Therefore, it is suggested that the group with high G6PD expression may have a weakened immune system.

[0088] From the results of the above-described examples, it can be said that the same tendency was observed regardless of the type of skin cancer.Therefore, it is suggested that regardless of the type of skin cancer, by using the expression level of G6PD as an index, it is possible to predict the prognosis of skin cancer, evaluate the effectiveness of immune checkpoint inhibitors, determine whether immune checkpoint inhibitors should be administered, evaluate the malignancy of skin cancer, predict the possibility of recurrence of skin cancer, evaluate the immune activity of skin cancer, and predict the possibility of Grade 3 or higher irAE caused by the administration of immune checkpoint inhibitors.

[0089] The present invention is not limited to the above-mentioned embodiment and examples. Various modifications within the scope of the claims and within the scope that can be easily conceived by a person skilled in the art are also included in the present invention.

Claims

1. 1. A method for prognosticating skin cancer, comprising: determining a correlation amount that correlates with the expression amount of glucose-6-phosphate dehydrogenase in a sample collected from the skin cancer patient; A high correlation indicates a poorer prognosis for the skin cancer than a low correlation. A method for predicting the prognosis of skin cancer.

2. 2. The method for predicting the prognosis of skin cancer according to claim 1, The skin cancer is at least one selected from the group consisting of Merkel cell carcinoma, malignant melanoma, squamous cell carcinoma, extramammary Paget's disease, and cutaneous angiosarcoma; A method for predicting the prognosis of skin cancer.

3. 3. The method for predicting the prognosis of skin cancer according to claim 2, The skin cancer includes Merkel cell carcinoma. A method for predicting the prognosis of skin cancer.

4. The method for predicting the prognosis of skin cancer according to any one of claims 1 to 3, The step of determining the correlation amount includes a step of determining the expression level of glucose-6-phosphate dehydrogenase by immunohistochemical staining of the sample. A method for predicting the prognosis of skin cancer.

5. The method for predicting the prognosis of skin cancer according to any one of claims 1 to 3, The step of determining the correlation amount includes a step of determining an expression level of mRNA encoding the glucose-6-phosphate dehydrogenase in the sample. A method for predicting the prognosis of skin cancer.

6. The method for predicting the prognosis of skin cancer according to any one of claims 1 to 3, The step of determining the correlation amount includes a step of measuring the activity of glucose-6-phosphate dehydrogenase in blood or serum as the sample. A method for predicting the prognosis of skin cancer.

7. 1. A method for evaluating the efficacy of an immune checkpoint inhibitor against skin cancer, comprising: determining a correlation amount that correlates with the expression amount of glucose-6-phosphate dehydrogenase in a sample collected from the skin cancer patient; When the correlation amount is small, the effectiveness of the immune checkpoint inhibitor is higher than when the correlation amount is large. method.

8. A method for determining whether or not to administer the immune checkpoint inhibitor to the patient, comprising the method of claim 7, indicating that, if the efficacy of the immune checkpoint inhibitor is evaluated to be high, the patient is administered or continues to be administered the immune checkpoint inhibitor; method.

9. 1. A method for assessing the aggressiveness of skin cancer, comprising: determining a correlation amount that correlates with the expression amount of glucose-6-phosphate dehydrogenase in a sample collected from the skin cancer patient; A large correlation amount indicates a high malignancy of the skin cancer compared to a small correlation amount. method.

10. A biomarker for predicting the prognosis of skin cancer, comprising: Glucose-6-phosphate dehydrogenase in a sample taken from a patient with skin cancer; A high expression level of the biomarker in the sample indicates a poorer prognosis of the skin cancer than a low expression level of the biomarker in the sample. Biomarkers.

11. A biomarker for evaluating the efficacy of an immune checkpoint inhibitor against skin cancer, comprising: Glucose-6-phosphate dehydrogenase in a sample taken from a patient with skin cancer; When the expression level of the biomarker in the sample is low, the effectiveness of the immune checkpoint inhibitor is higher than when the expression level is high. Biomarkers.

12. A biomarker for assessing the malignancy of skin cancer, comprising: Glucose-6-phosphate dehydrogenase in a sample taken from a patient with skin cancer; A high expression level of the biomarker in the sample indicates a high malignancy of the skin cancer compared to a low expression level of the biomarker in the sample. Biomarkers.

13. A biomarker for predicting recurrence of skin cancer, comprising: Glucose-6-phosphate dehydrogenase in a sample taken from a patient suffering from said skin cancer; a high expression level of the biomarker in the sample indicates a higher likelihood of recurrence of the skin cancer than a low expression level of the biomarker in the sample; Biomarkers.

14. A measurement kit comprising: The method is used for at least one of predicting the prognosis of skin cancer, evaluating the effectiveness of an immune checkpoint inhibitor against the skin cancer, evaluating the malignancy of the skin cancer, predicting the recurrence of the skin cancer, and evaluating the immune activity of the skin cancer, The present invention further comprises a substance capable of detecting an amount correlated with the expression level of glucose-6-phosphate dehydrogenase in a sample collected from a patient with skin cancer. Measuring kit.

15. The assay kit according to claim 14, The substance capable of detecting the correlation amount includes at least one selected from the group consisting of a substance capable of binding to the glucose-6-phosphate dehydrogenase or a fragment thereof, a substance capable of binding to a gene encoding the glucose-6-phosphate dehydrogenase, and glucose-6-phosphate. Measuring kit.

Images