Kit and diagnosis assistance method

Abstract

Provided is a method for diagnosing the prognosis and therapeutic effect for chemotherapy in patients with rheumatoid arthritis-associated lymphoproliferative disorder (RA-LPD), who have developed a lymphoproliferative disorder (LPD) during the treatment of rheumatoid arthritis (RA). This is a kit for diagnosing the prognosis and therapeutic effect for chemotherapy in rheumatic arthritis-associated lymphoproliferative disorder (RA-LPD) patients, who have developed LPD during RA treatment. The kit comprises: means for detecting an inositol 1,4,5-triphosphate receptor (ITPR) type 2 (ITPR2) gene mutation in a biological sample from a patient; and instructions for determining that the prognosis and therapeutic effect for chemotherapy are poor when an ITPR2 gene mutation is present in the biological sample from the patient. The LPD is diffuse large B-cell lymphoma (DLBCL). For the ITPR2 gene mutation, Chr12:rs26744460 is GT or GG. Use for the diagnosis of non-RA lymphoma patients is also possible.

Description

Kit and diagnostic aid method

[0001] The present invention relates to a kit for diagnosing the therapeutic effect of chemotherapy and prognosis of patients with rheumatoid arthritis-associated lymphoproliferative disorder (RA-LPD), who develop lymphoproliferative disorder (LPD) during treatment for rheumatoid arthritis (RA), and a diagnostic aid method therefor, and also to a kit for diagnosing the therapeutic effect of chemotherapy and prognosis of malignant lymphoma, even in non-RA patients, and a diagnostic aid method therefor.

[0002] RA patients are at high risk for developing lymphoma, with standardized incidence ratios (SIRs) reported to be 2-6 times higher. Recently, the number of reported cases of lymphoproliferative disorders (RA-LPDs) occurring during RA treatment has been increasing in Japan. LPDs are defined as "a state of excessive lymphocyte proliferation, not a single tumor, but encompassing everything from spontaneously resolving benign or reactive lymphocyte proliferations to true malignant lymphomas." LPDs occurring during immunosuppressive therapy for RA, such as methotrexate (MTX) or TNF inhibitors, are classified as other iatrogenic immunodeficiency-associated lymphoproliferative disorders (OIIA-LPDs) in the WHO classification. While intensive RA treatments using MTX and JAK inhibitors have dramatically improved outcomes in recent years, the possibility remains that intensive immunosuppressive therapy may increase the risk of developing LPD (Non-Patent Document 1).

[0003] Among malignant lymphoma cases in Japanese patients, diffuse large B-cell lymphoma (DLBCL) is the most common histological subtype. DLBCL belongs to the intermediate- to high-grade B-cell lymphoma category and accounts for 30-40% of non-Hodgkin's lymphomas. The standard treatment for DLBCL is R-CHOP therapy (Rituxan, cyclophosphamide, doxorubicin, vincristine, and prednisolone), but approximately 30% of cases show poor response to chemotherapy, resulting in a poor prognosis (Non-Patent Documents 2 and 3).

[0004] Furthermore, early administration of MTX has become the first choice for the treatment of RA, and it is expected that MTX-related LPD will increase in the future. When RA-LPD is diagnosed, remission may be achieved simply by discontinuing MTX. The standard treatment for RA-LPD in DLBCL, like that for regular lymphomas, is six courses of R-CHOP (Rituxan, cyclophosphamide, doxorubicin, vincristine, prednisolone) therapy; however, like regular lymphomas, chemotherapy is ineffective in approximately 20% of patients (Non-Patent Document 4).

[0005] Guidelines for the diagnosis and management of rheumatoid arthritis-associated lymphoproliferative disorders. 2022 Yodosha Guidelines for the Treatment of Hematopoietic Tumors, published by the Joint Working Group on RA-associated LPD (Japan Rheumatology Association, Japanese Society of Hematology, and Japanese Society of Pathology). Japanese Society of Hematology. http: / / www.jshem.or.jp / gui-hemali / index.html (accessed 2024-7-6). He MY, Kridel R. Treatment resistance in diffuse large B-cell lymphoma. Leukemia 2021; 35:2151-2165. Hoshida Y, Tsujii A, Ohshima S, Saeki Y, et al. Effect of recent antirheumatic drugs on features of rheumatoid arthritis-associated lymphoproliferative disorders. Arthritis rheumatology. 2024; 76:869-881. DOI 10.1002 / art.42809

[0006] The present invention has been made in consideration of these problems, and aims to provide a method for diagnosing the therapeutic effect of chemotherapy and prognosis in patients with RA-LPD, who develop LPD during RA treatment, and also to provide a method for diagnosing the therapeutic effect of chemotherapy for malignant lymphoma in non-RA patients.

[0007] The kit of the present invention is a kit for diagnosing the therapeutic effect of chemotherapy and prognosis of a patient with RA-LPD, who develops LPD during RA treatment, and is characterized by comprising a means for detecting an inositol 1,4,5-triphosphate receptor (ITPR) type 2 (ITPR2) gene mutation in a biological sample from the patient, and instructions for determining that the therapeutic effect and prognosis are poor if the patient's biological sample contains an ITPR2 gene mutation. Note that the human genome version used herein is based on hg19 (or GRCh37).

[0008] The kit of the present invention is a kit for diagnosing the therapeutic effect of chemotherapy in a patient with malignant lymphoma, and is characterized by comprising: a means for detecting an inositol 1,4,5-triphosphate receptors (ITPR) type 2 (ITPR2) gene mutation in a biological sample of the patient; and instructions for determining that the therapeutic effect of chemotherapy and the prognosis are poor if an ITPR2 gene mutation is present in the biological sample of the patient.

[0009] According to the present invention, it is possible to accurately diagnose the therapeutic effect of chemotherapy and the prognosis of RA-LPD patients.

[0010] This is a Manhattan plot of a GWAS analysis showing the treatment effect of RA-LPD (progressive disease vs. non-PD). This is a locus plot of a GWAS analysis showing the treatment effect of RA-LPD (PD vs. non-PD). a) Kaplan Meier curves for relapse-free period after treatment for RA-LPD at Chr12:rs26743031, with AA vs. GA or GG in order from top to bottom. b) Kaplan Meier curves for relapse-free period after treatment for RA-LPD at Chr12:rs26743031, with AA vs. GA vs. GG in order from top to bottom. c) Kaplan Meier curves for overall survival for RA-LPD at Chr12:rs26743031, with AA vs. GA or GG in order from top to bottom. d) Kaplan Meier curves for overall survival for RA-LPD at Chr12:rs26743031, with AA vs. GA vs. GG in order from top to bottom. e) Multivariate analysis forest plot for post-treatment relapse-free factors for RA-LPD. f) Multivariate analysis forest plot for post-treatment prognostic factors for RA-LPD. a) Kaplan Meier curves for post-treatment relapse-free periods for RA-LPD treated with R-CHOP therapy at Chr12:rs26743031, showing AA vs. GA or GG from top to bottom. b) Kaplan Meier curves for post-treatment relapse-free periods for RA-LPD treated with R-CHOP therapy at Chr12:rs26743031, showing AA vs. GA vs. GG from top to bottom. c) Kaplan Meier curves for overall survival for RA-LPD treated with R-CHOP therapy at Chr12:rs26743031, showing AA vs. GA or GG from top to bottom. d) Kaplan Meier curves for overall survival in RA-LPD patients receiving R-CHOP therapy at Chr12:rs26743031, with AA vs. GA vs. GG from top to bottom. e) Multivariate analysis forest plot of post-treatment recurrence-free factors in RA-LPD patients receiving R-CHOP therapy. f) Multivariate analysis forest plot of post-treatment prognostic factors in RA-LPD patients receiving R-CHOP therapy.a) Kaplan Meier curves for post-treatment relapse-free intervals for RA-LPD at Chr12:rs26744460, with TT vs. GT or GG from top to bottom. b) Kaplan Meier curves for post-treatment relapse-free intervals for RA-LPD at Chr12:rs26744460, with TT vs. GT vs. GG from top to bottom. c) Kaplan Meier curves for overall survival for RA-LPD at Chr12:rs26744460, with TT vs. GT or GG from top to bottom. d) Kaplan Meier curves for overall survival for RA-LPD at Chr12:rs26744460, with TT vs. GT vs. GG from top to bottom. e) Multivariate analysis forest plot for post-treatment relapse-free factors for RA-LPD. f) Multivariate analysis forest plot for post-treatment prognostic factors for RA-LPD. a) Kaplan Meier curves for post-treatment relapse-free intervals in RA-LPD patients receiving R-CHOP therapy at Chr12:rs26744460, with TT vs. GT at the top. b) Kaplan Meier curves for overall survival in RA-LPD patients receiving R-CHOP therapy at Chr12:rs26744460, with TT vs. GT at the top. c) Multivariate forest plots for post-treatment relapse-free intervals in RA-LPD patients receiving R-CHOP therapy. d) Multivariate forest plots for post-treatment prognostic factors in RA-LPD patients receiving R-CHOP therapy. a) ITPR2 mRNA expression in RA-LPD patients receiving GG vs. GT vs. TT at Chr12:rs26744460. b) ITPR2 mRNA expression in RA-LPD patients receiving GG or GT vs. TT at Chr12:rs26744460. a) Kaplan Meier curves of overall survival for DLBCL at Chr12:rs26734980, with AA vs. TA from top to bottom. b) Kaplan Meier curves of recurrence-free period after treatment for DLBCL at Chr12:rs26734980, with AA vs. TA from top to bottom.a) Fisher's exact test results for the correlation between 15 SNPs in the ITPR2 region and treatment-resistant cases in non-RA patients with DLBCL. b) The proportion of genetic polymorphisms in DLBCL CR (complete remission) and PD cases in non-RA patients with DLBCL at Chr12.26744269. c) Kaplan Meier curves of overall survival for DLBCL at Chr12.26744269, from top to bottom: TT vs. TC or CC. d) Kaplan Meier curves of relapse-free period after treatment for DLBCL at Chr12.26744269, from top to bottom: TT vs. TC or CC.

[0011] Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. However, these embodiments are intended to facilitate understanding of the principles of the present invention, and the scope of the present invention is not limited to the following embodiments. Other embodiments in which a person skilled in the art appropriately replaces the configuration of the following embodiments are also included in the scope of the present invention.

[0012] The kit of the present invention is a kit for diagnosing the therapeutic effect of chemotherapy and the prognosis of a patient with RA-LPD, who develops LPD during RA treatment.

[0013] RA is a systemic inflammatory disease of unknown cause, with polysynovial arthritis as its primary lesion. While most cases are chronic, progression can lead to joint destruction and deformation, resulting in joint dysfunction. While there is no established theory about the mechanism by which LPD develops during RA treatment, it is thought that the administration of RA medications induces an immunosuppressive state, leading to the onset of LPD.

[0014] The kit of the present invention includes a means for detecting an inositol 1,4,5-triphosphate receptors (ITPR) type 2 (ITPR2) gene mutation in a patient's biological sample, and instructions for determining that the patient's therapeutic response to chemotherapy and prognosis are poor if the patient's biological sample contains an ITPR2 gene mutation.

[0015] The means for detecting mutations in the inositol 1,4,5-triphosphate receptors (ITPR) type 2 (ITPR2) gene are not particularly limited, and any of the known SNP detection methods can be used to detect genetic polymorphisms. Classical detection methods include, for example, a method in which genomic DNA extracted from a patient's cells or the like is used as a sample, and a nucleic acid containing a different base sequence of about 15 to about 500 bases including the base at the genetic polymorphism site is used as a probe, and hybridization is performed under precise stringency control to detect only the sequence that is completely complementary to the probe; and a method in which a mixed probe is used in which one of the nucleic acids in which the base at the polymorphism site is substituted with another base is labeled, and the other is unlabeled, and hybridization is performed while gradually decreasing the reaction temperature from the denaturation temperature, allowing the sequence that is completely complementary to one probe to hybridize first and preventing cross-reaction with a mismatched probe.

[0016] Preferably, the detection of gene polymorphisms can be carried out by, for example, TaqMan PCR, RFLP, PCR-SSCP, ASO hybridization, direct sequencing, ARMS, denaturing gradient gel electrophoresis, RNase A cleavage, chemical cleavage, DOL, Invader, MALDI-TOF / MS, TDI, molecular beacon, dynamic allele-specific hybridization, padlock probe, UCAN, nucleic acid hybridization using a DNA chip or DNA microarray, and ECA.

[0017] Furthermore, the means for detecting the inositol 1,4,5-triphosphate receptors (ITPR) type 2 (ITPR2) gene mutation is not particularly limited, but examples include quantitative PCR (including real-time PCR). This quantitative PCR method uses a primer set capable of amplifying the sequence of the ITPR2 gene mutation. As long as it is capable of detecting the ITPR2 gene mutation, it is not particularly limited, and conventional quantitative PCR methods such as fluorescent probe methods (e.g., TaqMan® probe method), agarose electrophoresis, and SYBR Green can be used. In quantitative PCR, a primer set refers to a combination of primers (polynucleotides) capable of amplifying the sequence of the ITPR2 gene mutation. The primers are not particularly limited as long as they are capable of amplifying the sequence of the ITPR2 gene mutation, but an example is a primer set consisting of a forward primer and a reverse primer. The primer set can be obtained by chemical synthesis or the like using methods well known in the art based on the sequence information.

[0018] Instead of detecting an ITPR2 gene mutation, a means for detecting an ITPR2 gene mutation can be to detect a mutation in an ITPR2-related gene, such as, but not limited to, ITPR1, ITPR3, PLCG2, CALM3, or PSEN2.

[0019] Alternatively, instead of detecting ITPR2 gene mutations, it is also possible to detect mutations in ITPR2-related pathway genes, such as ITPR1, ITPR3, PLCG2, CALM3, CALML3, CALML4, CALML5, CALML6, and PSEN2.

[0020] The instructions for determining that the therapeutic effect on chemotherapy and prognosis are poor when there is an ITPR2 gene mutation in the patient's biological sample can, for example, indicate that the therapeutic effect on chemotherapy and prognosis are poor when there is a mutation in the ITPR2 gene in the patient's biological sample but no mutation in the ITPR2 gene in the control biological sample.

[0021] LPD that develops during RA treatment is not particularly limited, but examples include diffuse large B-cell lymphoma (DLBCL) and Hodgkin's lymphoma.

[0022] In the case of LPD that develops during RA treatment, examples of ITPR2 gene mutations include Chr12:rs26743031 being GA or GG, and Chr12:rs26744460 being GT or GG.

[0023] When LPD that develops during RA treatment is chronic large B-cell lymphoma (DLBCL), the ITPR2 gene mutation may be TA or TT at Chr12:26734980.

[0024] In the case of LPD that develops during RA treatment, for example, chemotherapy includes R-CHOP therapy (Rituxan, cyclophosphamide, doxorubicin, vincristine, prednisolone), but if the aforementioned ITPR2 gene mutation is present, it can be determined that the treatment effect and prognosis are poor.

[0025] In the case of LPD that develops during RA treatment, for example, chemotherapy includes ABVD therapy (doxorubicin, bleomycin, vinblastine, and dacarbazine), but if the aforementioned ITPR2 gene mutation is present, it can be determined that the treatment effect and prognosis are poor.

[0026] The biological sample is not particularly limited and includes blood, serum, tears, saliva, urine, biopsy tissue, etc. from the patient, and for example, blood can be preferably used.

[0027] The kit of the present invention may further include other reagents, reaction vessels, instructions, etc., depending on the specific method to be carried out.

[0028] The kit of the present invention is a kit for diagnosing the therapeutic effect of chemotherapy in patients with malignant lymphoma, and is characterized by comprising a means for detecting inositol 1,4,5-triphosphate receptors (ITPR) type 2 (ITPR2) gene mutations in a biological sample from the patient, and instructions for determining that the therapeutic effect of chemotherapy and prognosis are poor if the patient's biological sample contains an ITPR2 gene mutation. The malignant lymphoma patients herein include patients who do not have rheumatoid arthritis.

[0029] The malignant lymphoma is not particularly limited, but may be, for example, non-Hodgkin's lymphoma, such as follicular lymphoma, MALT lymphoma, chronic large B-cell lymphoma (DLBCL), mantle cell lymphoma, peripheral T-cell lymphoma-unspecified type, angioimmunoblastic T-cell lymphoma, anaplastic large cell lymphoma, extranodal NK / T-cell lymphoma (nasal type), Burkitt's lymphoma, lymphoblastic lymphoma, and adult T-cell leukemia / lymphoma, preferably DLBCL.

[0030] In the case of DLBCL, ITPR2 gene mutations are, for example, Chr12.26740364 AG or GG, Chr12.26755546 CT or TT, Chr12.26744269 TC or CC, Chr12:26730914 CT or TT, Chr12:26730959 CT or TT, Chr12:26739021 AG, ​​Chr12:26744097 AG, Chr12:26746089 GA or AA, Chr12:26744269 TC or CC, Chr12:26750581 TA or AA, or Chr12:26756713 TA or TT.

[0031] Furthermore, the diagnostic support method of the present invention is a diagnostic support method for diagnosing the therapeutic effect of chemotherapy in patients with RA-LPD who develop LPD during RA treatment, and is characterized by comprising the steps of: detecting an inositol 1,4,5-triphosphate receptors (ITPR) type 2 (ITPR2) gene mutation in a biological sample from the patient; and determining that the therapeutic effect and prognosis are poor if an ITPR2 gene mutation is present in the patient's biological sample.

[0032] Furthermore, the diagnostic support method of the present invention is a diagnostic support method for diagnosing the therapeutic effect of chemotherapy and prognosis of a patient with malignant lymphoma, and is characterized by comprising the steps of detecting an inositol 1,4,5-triphosphate receptors (ITPR) type 2 (ITPR2) gene mutation in a biological sample from the patient, and determining that the therapeutic effect and prognosis are poor if an ITPR2 gene mutation is present in the patient's biological sample.

[0033] To explore genes related to the pathogenesis of RA-LPD, we conducted a multi-institutional collaborative study centered on hospitals of the National Hospital Organization (NHO), and performed comprehensive integrated analysis using GWAS analysis and RNA transcriptome analysis.

[0034] First, we performed a comprehensive GWAS analysis suitable for Japanese people using the Japonica Array on 238 RA patients with LPD. The probes used were 659,636 SNPs, and imputation analysis was performed on approximately 4 million SNPs.

[0035] Next, we performed comprehensive RNA transcriptome analysis on blood samples from 191 RA patients with LPD to examine gene expression at the mRNA level. Figure 1 shows a Manhattan plot of the GWAS analysis showing the treatment effect in RA-LPD (progressive disease vs. non-PD). As shown in Figure 1, genetic information related to the pathogenesis of RA-LPD was obtained.

[0036] Figure 2 is a GWAS analysis locus plot showing the treatment effect (PD vs. non-PD) in RA-LPD. Table 1 shows the results of the GWAS analysis of gene polymorphisms in RA-LPD whose association with PD (progressive disease) exceeded the genome-wide level. Table 1 and Figure 2 show that 19 SNPs in the ITPR2 region, including the enhancer region, were associated with RA-LPD treatment resistance above the genome-wide significance level.

[0037]

[0038] Rheumatoid arthritis-associated lymphoproliferative disorders (RA-LPD) are lymphoproliferative disorders that occur in patients with rheumatoid arthritis.

[0039] Figure 3a) shows the Kaplan Meier curves for post-treatment relapse-free intervals for RA-LPD (AA vs. GA or GG) for Chr12:rs26743031. Figure 3b) shows the Kaplan Meier curves for post-treatment relapse-free intervals for RA-LPD (AA vs. GA vs. GG) for Chr12:rs26743031. Figure 3c) shows the Kaplan Meier curves for overall survival for RA-LPD (AA vs. GA or GG) for Chr12:rs26743031. Figure 3d) shows the Kaplan Meier curves for overall survival for RA-LPD (AA vs. GA vs. GG) for Chr12:rs26743031. Figure 3e) shows the multivariate forest plot for post-treatment relapse-free factors for RA-LPD. Figure 3f) shows the multivariate forest plot for post-treatment prognostic factors for RA-LPD.

[0040] As shown in Figure 3, the SNP that met the highest genome-wide significance level was Chr12:rs26743031 (p = 4.68E-10). Compared to AA, the GA or GG polymorphism was associated with a significantly shorter period until recurrence after treatment for RA-LPD, and also showed a correlation with poor prognosis. Multivariate analysis showed that this could be an indicator of treatment resistance and prognosis for RA-LPD.

[0041] As shown in Figure 4, even in RA-LPD patients who received R-CHOP therapy, polymorphisms of the above SNPs were associated with a significantly shorter time to recurrence after treatment and a correlation with poor prognosis was observed, demonstrating that they can be used as indicators of treatment resistance and prognosis in multivariate analysis.

[0042] Figure 5a) shows the Kaplan Meier curves for post-treatment relapse-free intervals for RA-LPD (TT vs. GT or GG) for Chr12:rs26744460. Figure 5b) shows the Kaplan Meier curves for post-treatment relapse-free intervals for RA-LPD (TT vs. GT vs. GG) for Chr12:rs26744460. Figure 5c) shows the Kaplan Meier curves for overall survival for RA-LPD (TT vs. GT or GG) for Chr12:rs26744460. Figure 5d) shows the Kaplan Meier curves for overall survival for RA-LPD (TT vs. GT vs. GG) for Chr12:rs26744460. Figure 5e) shows the multivariate forest plot for post-treatment relapse-free factors for RA-LPD. Figure 5f) shows the multivariate forest plot for post-treatment prognostic factors for RA-LPD.

[0043] As shown in Figure 5, for Chr12:rs26744460, the GT or GG polymorphism was associated with a significantly shorter time to recurrence after treatment compared to the TT polymorphism, and also showed a correlation with poor prognosis. Multivariate analysis showed that this polymorphism can be an indicator of treatment resistance and prognosis in DLBCL.

[0044] As shown in Figure 6, even in RA-LPD patients who underwent R-CHOP therapy, the polymorphisms of the above SNPs were correlated with a significantly shorter time to recurrence after treatment and a poor prognosis, and multivariate analysis showed that they can be used as indicators of treatment resistance and prognosis.

[0045] Figure 7a) shows ITPR2 mRNA expression in RA-LPD patients with GG vs. GT vs. TT at Chr12:rs26744460. Figure 7b) shows ITPR2 mRNA expression in RA-LPD patients with GG or GT vs. TT at Chr12:rs26744460.

[0046] As shown in Figure 7, a significant difference in ITPR2 expression levels in RA-LPD was observed in the Chr12:rs26744460 region, which met the genome-wide significance level of p = 8.61E-08 for RA-LPD overall and 3.75E-08 for DLBCL.

[0047] Diffuse large B-cell lymphoma (DLBCL) is a type of malignant lymphoma characterized by diffuse proliferation of large B-cell tumor cells with nuclei that are more than twice the size of small lymphocytes or are equal to or larger than the nuclei of normal histiocytes.

[0048] Figure 8a) Kaplan Meier curve of overall survival for DLBCL AA vs TA at Chr12:rs26734980.

[0049] Figure 8b) shows the Kaplan Meier curve AA vs TA of the relapse-free period after treatment for DLBCL at Chr12:rs26734980.

[0050] As shown in Figure 8, for Chr12:rs26734980, the TA or TT polymorphism was associated with a significantly shorter period until recurrence after treatment compared to the AA polymorphism, and also showed a correlation with poor prognosis, indicating that it can be an indicator of treatment resistance and prognosis for DLBCL.

[0051] Next, we investigated the correlation between ITPR2 gene polymorphisms and treatment resistance (PD) in non-RA patients who developed malignant lymphoma.

[0052] Specifically, SNP analysis was performed on 15 SNPs in the ITPR2 region using a total of 123 cases (blood samples (25 patients) and FFPE samples (98 patients)) of DLBCL patients, including 67 PD cases and 56 CR cases.

[0053] Figure 9(a) shows the Fisher exact test results for the correlation between 15 SNPs in the ITPR2 region and treatment-resistant cases in non-RA patients with DLBCL. As shown in Figure 9(a), 12 of the 15 SNPs were correlated with PD cases. Three SNPs (Chr12.26740364, Chr12.26744269, Chr12.26755546) were correlated with prognosis. Specifically, at Chr12.26740364, the AG or GG polymorphism was correlated with poor prognosis compared with the AA polymorphism. At Chr12.26755546, the CT or TT polymorphism was correlated with poor prognosis compared with the CC polymorphism.

[0054] Figure 9b) shows the proportion of CR and PD cases of DLBCL at Chr12.26744269 in non-RA patients who developed DLBCL. As shown in Figure 9b), TC or CC polymorphisms were significantly associated with poor prognosis compared with TT.

[0055] Figure 9c) shows the Kaplan-Meier curve of overall survival for DLBCL (TT vs. TC or CC) for Chr12.26744269. As shown in Figure 9c), the TC or CC polymorphisms were correlated with poor prognosis after treatment for DLBCL compared with the TT polymorphism, demonstrating their potential as prognostic indicators for DLBCL.

[0056] Figure 9d) shows the Kaplan-Meier curve of the relapse-free period after treatment for DLBCL (TT vs. TC or CC) for Chr12.26744269. As shown in Figure 9d), the TC or CC polymorphisms were correlated with a significantly shorter period until relapse after treatment for DLBCL compared to TT, indicating that they are indicators of treatment resistance for DLBCL.

[0057] It can be used to treat RA-LPD and malignant lymphoma in non-RA patients.

Claims

1. A kit for diagnosing the therapeutic effect of chemotherapy and prognosis of a patient with rheumatoid arthritis-associated lymphoproliferative disease who develops lymphoproliferative disease during treatment for rheumatoid arthritis, the kit comprising: a means for detecting inositol 1,4,5-triphosphate receptors (ITPR) type 2 (ITPR2) gene mutations in a biological sample from the patient; and instructions for determining that the therapeutic effect of chemotherapy and prognosis are poor if an ITPR2 gene mutation is present in the patient's biological sample.

2. The kit according to claim 1, further comprising a means for detecting a mutation in an ITPR2-related gene instead of detecting an ITPR2 gene mutation in the patient's biological sample.

3. The kit according to claim 2, wherein the ITPR2-related gene is any one of ITPR1, ITPR3, PLCG2, CALM3, or PSEN2.

4. The kit according to claim 1, characterized in that the lymphoproliferative disease is diffuse large B-cell lymphoma (DLBCL).

5. The kit according to claim 4, characterized in that if an ITPR2 gene mutation is present in the patient's biological sample, it is determined that the therapeutic effect of R-CHOP therapy (Rituxan, cyclophosphamide, doxorubicin, vincristine, prednisolone) and the prognosis are poor.

6. The kit described in claim 4, characterized in that the ITPR2 gene mutation Chr12:rs26743031 is GA or GG.

7. The kit described in claim 4, characterized in that the ITPR2 gene mutation is Chr12:rs26744460 GT or GG.

8. The kit described in claim 4, characterized in that the ITPR2 gene mutation is TA or TT at Chr12:26734980.

9. The kit according to claim 1, wherein the lymphoproliferative disease is rheumatoid arthritis-associated lymphoproliferative disease.

10. The kit described in claim 9, characterized in that if an ITPR2 gene mutation is present in the patient's biological sample, it is determined that the therapeutic effect and prognosis of ABVD therapy (doxorubicin, bleomycin, vinblastine, dacarbazine) are poor.

11. The kit described in claim 9, characterized in that the ITPR2 gene mutation Chr12:rs26743031 is GA or GG.

12. A kit for diagnosing the therapeutic effect of chemotherapy in a patient with malignant lymphoma, comprising: a means for detecting an inositol 1,4,5-triphosphate receptors (ITPR) type 2 (ITPR2) gene mutation in a biological sample of the patient; and instructions for determining that the therapeutic effect of chemotherapy and the prognosis are poor if an ITPR2 gene mutation is present in the biological sample of the patient.

13. The kit according to claim 12, further comprising a means for detecting a mutation in an ITPR2-related gene instead of detecting an ITPR2 gene mutation in the patient's biological sample.

14. The kit according to claim 13, wherein the ITPR2-related gene is any one of ITPR1, ITPR3, PLCG2, CALM3, and PSEN2.

15. The kit according to claim 12, wherein the malignant lymphoma is diffuse large B-cell lymphoma (DLBCL).

16. The kit of claim 12, wherein the malignant lymphoma is follicular lymphoma, MALT lymphoma, mantle cell lymphoma, peripheral T-cell lymphoma, unspecified type, angioimmunoblastic T-cell lymphoma, anaplastic large cell lymphoma, extranodal NK / T-cell lymphoma (nasal type), Burkitt lymphoma, lymphoblastic lymphoma, or adult T-cell leukemia / lymphoma.

17. The kit described in claim 15, characterized in that if an ITPR2 gene mutation is present in the patient's biological sample, it is determined that the therapeutic effect of R-CHOP therapy (Rituxan, cyclophosphamide, doxorubicin, vincristine, prednisolone) and the prognosis are poor.

18. The kit described in claim 15, characterized in that the ITPR2 gene mutation is TC or CC at Chr12:rs26744269.

19. The kit described in claim 15, characterized in that the ITPR2 gene mutation is Chr12:rs26740364 AG or GG.

20. The kit described in claim 15, characterized in that the ITPR2 gene mutation Chr12:rs26755546 is CT or TT.

21. The kit described in claim 15, characterized in that the ITPR2 gene mutation is CT or TT at Chr12:26730914.

22. The kit described in claim 15, characterized in that the ITPR2 gene mutation is CT or TT at Chr12:26730959.

23. The kit described in claim 15, characterized in that the ITPR2 gene mutation is AG at Chr12:26739021.

24. The kit described in claim 15, characterized in that the ITPR2 gene mutation is AG at Chr12:26744097.

25. The kit described in claim 15, characterized in that the ITPR2 gene mutation is GA or AA at Chr12:26746089.

26. The kit described in claim 15, characterized in that the ITPR2 gene mutation is TC or CC at Chr12:26744269.

27. The kit described in claim 15, characterized in that the ITPR2 gene mutation is TA or AA at Chr12:26750581.

28. The kit described in claim 15, characterized in that the ITPR2 gene mutation is TA or TT at Chr12:26756713.

29. A diagnostic auxiliary method for diagnosing the therapeutic effect of chemotherapy and prognosis of a patient with rheumatoid arthritis-associated lymphoproliferative disease who develops lymphoproliferative disease during treatment for rheumatoid arthritis, the diagnostic auxiliary method comprising the steps of: detecting an inositol 1,4,5-triphosphate receptors (ITPR) type 2 (ITPR2) gene mutation in a biological sample from the patient; and determining that the therapeutic effect and prognosis are poor if an ITPR2 gene mutation is present in the patient's biological sample.

30. The diagnostic aid method according to claim 29, further comprising a means for detecting a mutation in an ITPR2-related gene instead of detecting an ITPR2 gene mutation in the patient's biological sample.

31. The diagnostic assistance method according to claim 30, wherein the ITPR2-related gene is any one of ITPR1, ITPR3, PLCG2, CALM3, and PSEN2.

32. The diagnostic aid method according to claim 29, wherein the lymphoproliferative disease is diffuse large B-cell lymphoma (DLBCL).

33. The diagnostic auxiliary method according to claim 32, characterized in that if an ITPR2 gene mutation is present in the patient's biological sample, it is determined that the therapeutic effect of R-CHOP therapy (rituximab, chlorophosphamide, doxorubicin, vincristine, prednisolone) and the prognosis are poor.

34. The diagnostic auxiliary method described in claim 32, characterized in that the ITPR2 gene mutation is GA or GG at Chr12:rs26743031.

35. The kit diagnostic auxiliary method described in claim 32, characterized in that the ITPR2 gene mutation is Chr12:rs26744460 is GT or GG.

36. The diagnostic auxiliary method described in claim 32, characterized in that the ITPR2 gene mutation is TA or TT at Chr12:26734980.

37. The diagnostic assistance method according to claim 29, wherein the lymphoproliferative disease is a rheumatoid arthritis-associated lymphoproliferative disease.

38. The diagnostic auxiliary method described in claim 37, characterized in that if an ITPR2 gene mutation is present in the patient's biological sample, it is determined that the therapeutic effect and prognosis of ABVD therapy (doxorubicin, bleomycin, vinblastine, dacarbazine) are poor.

39. The kit diagnostic auxiliary method described in claim 37, characterized in that the ITPR2 gene mutation is GA or GG at Chr12:rs26743031.

40. A diagnostic auxiliary method for diagnosing the therapeutic effect of chemotherapy and prognosis of a patient with malignant lymphoma, comprising the steps of: detecting an inositol 1,4,5-triphosphate receptors (ITPR) type 2 (ITPR2) gene mutation in a biological sample of the patient; and determining that the therapeutic effect and prognosis are poor if an ITPR2 gene mutation is present in the patient's biological sample.

41. The diagnostic aid method according to claim 40, further comprising a means for detecting a mutation in an ITPR2-related gene instead of detecting an ITPR2 gene mutation in the patient's biological sample.

42. The diagnostic assistance method according to claim 41, wherein the ITPR2-related gene is any one of ITPR1, ITPR3, PLCG2, CALM3, or PSEN2.

43. The diagnostic assistance method according to claim 40, wherein the malignant lymphoma is diffuse large B-cell lymphoma (DLBCL).

44. The diagnostic assistance method of claim 40, wherein the malignant lymphoma is follicular lymphoma, MALT lymphoma, mantle cell lymphoma, peripheral T-cell lymphoma-not otherwise specified, angioimmunoblastic T-cell lymphoma, anaplastic large cell lymphoma, extranodal NK / T-cell lymphoma (nasal type), Burkitt lymphoma, lymphoblastic lymphoma, or adult T-cell leukemia / lymphoma.

45. The diagnostic auxiliary method described in claim 43, characterized in that if an ITPR2 gene mutation is present in the patient's biological sample, it is determined that the therapeutic effect of R-CHOP therapy (Rituxan, cyclophosphamide, doxorubicin, vincristine, prednisolone) and the prognosis are poor.

46. ​​The diagnostic auxiliary method described in claim 43, characterized in that the ITPR2 gene mutation is TC or CC at Chr12:rs26744269.

47. The diagnostic auxiliary method described in claim 43, characterized in that the ITPR2 gene mutation is Chr12:rs26740364 AG or GG.

48. The diagnostic auxiliary method described in claim 43, characterized in that the ITPR2 gene mutation is CT or TT at Chr12:rs26755546.

49. The diagnostic auxiliary method described in claim 43, characterized in that the ITPR2 gene mutation is CT or TT at Chr12:26730914.

50. The diagnostic auxiliary method described in claim 43, characterized in that the ITPR2 gene mutation is CT or TT at Chr12:26730959.

51. The diagnostic auxiliary method described in claim 43, characterized in that the ITPR2 gene mutation is AG at Chr12:26739021.

52. The diagnostic auxiliary method described in claim 43, characterized in that the ITPR2 gene mutation is AG at Chr12:26744097.

53. The diagnostic auxiliary method described in claim 43, characterized in that the ITPR2 gene mutation is GA or AA at Chr12:26746089.

54. The diagnostic auxiliary method described in claim 43, characterized in that the ITPR2 gene mutation is TC or CC at Chr12:26744269.

55. The diagnostic auxiliary method described in claim 43, characterized in that the ITPR2 gene mutation is TA or AA at Chr12:26750581.

56. The diagnostic auxiliary method described in claim 43, characterized in that the ITPR2 gene mutation is TA or TT at Chr12:26756713.

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