WO2022017435A1 - SERIES OF BIOMARKERS OF CONSENSUS SEQUENCES IN CDR3 SEQUENCE OF TCR-β CHAIN AND APPLICATION THEREOF - Google Patents

Abstract

A series of biomarkers of consensus sequences in a CDR3 sequence of a TCR-β chain and an application thereof. Provided is a set of biomarkers, characterized in that the biomarkers are consensus sequences in the CDR3 sequence of a T cell receptor (TCR) β chain. Preferably, the amino acid sequence of the biomarkers comprises RGG. The biomarkers provided may be used to predict the clinical response effect of PD1/PDL1 pathway immune checkpoint inhibitors to tumors, especially Hodgkin's lymphoma. High-throughput sequencing and statistical analysis are performed on the CDR3 sequence of the TCR-β chain of Hodgkin's lymphoma patients who are given the PD1/PDL1 pathway immune checkpoint inhibitors, and a dominant sequence is determined. By means of the enrichment analysis of the dominant sequence, it is helpful to monitor and predict the response and irAE of the patient after checkpoint immunotherapy.

Description

Biomarkers of consensus sequences in CDR3 sequences of a series of TCR-β chains and their applications

Priority and related applications

This application claims the priority of Chinese patent application 202010710792.7 filed on July 22, 2020, entitled "Biomarkers of Consensus Sequences in CDR3 Sequences of a Series of TCR-β Chains and Their Applications", which includes the appendix The entire contents of this application are incorporated by reference.

technical field

The invention belongs to the field of biomedicine and relates to the research of clinical biomarkers, in particular to a series of biomarkers of consensus sequences in the CDR3 sequences of TCR-beta chains and applications thereof.

Background technique

In recent years, immunotherapy has become a new mode of tumor treatment following surgery, radiotherapy, chemotherapy and targeted therapy. Tumor immunotherapy kills or controls tumors by regulating and activating the immune system. In the research progress of tumor immunotherapy, immune checkpoint inhibitors, especially programmed cell death-1 (PD-1)/programmed cell death ligand-1 (programmed cell death ligand) -1, PD-L1) inhibitors have received the most attention. Studies have shown that the expression levels of immune checkpoints such as PD-1/PD-L1 are associated with poor tumor prognosis and the effect of tumor immunotherapy. Antibodies to PD-1 and PD-L1 targeting immune checkpoints have become a new generation of cancer treatments.

Lymphoma is one of the common malignant tumors in my country. Classical Hodgkin's lymphoma (cHL) is a B-cell malignant lymphoma, which occurs mostly in young and middle-aged people. It is characterized by chromosome 9p24.1 mutation and PD. -1 for overexpression. Classical Hodgkin lymphoma (cHL) is a tumor type that responds well to PD-1 antibody therapy.

Sintilimab, a PD-1 checkpoint inhibitor, has demonstrated efficacy in relapsed/refractory cHL in a preliminary analysis of the ORIENT-1 study. The safety and efficacy of sintilimab after extended follow-up were evaluated in the Phase II trial of the ORIENT-1 study. The clinical overall response rate (ORR) of PD-1 and PD-L1 antibody drugs in relapsed/resistant (r/r) cHL can be as high as 66%-80% ^[1-4] . However, the response efficiency of each individual patient remains highly variable and difficult to predict. Moreover, 68%-93% of patients will experience immune-related adverse events (irAEs) after checkpoint inhibitor therapy, of which 12%-18% of patients achieve grade 3-4 side effects ^{[5] -6]} . Therefore, clinical markers that can well predict clinical response or immune-related side effects need to be developed.

T cell receptors (TCRs) are heterodimers responsible for recognizing antigens presented by the major histocompatibility complex (MHC). Most TCRs are composed of two polypeptide chains ( namely α chain and β chain), and each polypeptide chain contains three highly variable complementarity determining regions (CDRs), namely CDR1, CDR2 and CDR3, respectively. While the antigenic specificity of TCR is largely determined by the CDR3 of the T cell receptor beta chain, as it is generated by the rearrangement of various VDJ gene segments and random addition of junction site sequences, CDR3 is responsible for directly interacting with Binding of polypeptides presented by MHC. Since CDR3 interacts most closely with antigenic peptides, the diversity of CDR3 amino acid sequences provides an indicator of T cell diversity in the T cell repertoire ^[7-9] .

Clinical efficacy and irAEs are caused by antigen-specific T cell responses. In the prior art, neither PD-1 antibodies have been reported to produce highly enriched CDR3 sequences of TCR-β chains in patients with Hodgkin's lymphoma after clinical treatment, nor has it been reported that the enriched sequences will be CDR3 sequences. In-depth study of biological functions.

SUMMARY OF THE INVENTION

Invention to solve problem

In view of the problems existing in the prior art, the present application provides a series of biomarkers sharing the CDR3 sequence of the TCR-β chain and their application in predicting the clinical response of PD-1 antibody to Hodgkin lymphoma.

solution to the problem

In view of the above-mentioned problems in the prior art, the present inventors have conducted in-depth research and repeated tests to determine the level of TCR- The CDR3 sequence of the beta chain is subjected to high-throughput sequencing, the highly enriched sequence is analyzed, and the relationship between the highly enriched sequence and the patient's T cells for the treatment is studied in combination with clinical observations, thereby completing the present invention. That is, the present invention is as follows:

In the first aspect of the present invention, a set of biomarkers is provided, characterized in that, the biomarkers are the consensus amino acid sequence in the CDR3 sequence of T cell receptor (TCR) beta chain; The marker contains RGG in its amino acid sequence.

In some specific embodiments, the biomarkers respectively comprise the amino acid sequence shown in any one of SEQ ID NO: 1 to SEQ ID NO: 9; preferably, the biomarkers respectively comprise SEQ ID NO: The amino acid sequence shown in any one of 1-2, SEQ ID NO: 4-6 or SEQ ID NO: 7; more preferably, the biomarkers respectively comprise the amino acid sequences in SEQ ID NO: 2 or SEQ ID NO: 6 amino acid sequence shown.

In the second aspect of the present invention, there is provided the use of the biomarkers described in the first aspect of the present invention in the preparation of predictive PD1/PDL1 pathway immune checkpoint inhibitors or PD1/PDL1 pathway immune checkpoint inhibitor combination therapeutic agents. Application of the kit for tumor response effects.

In the third aspect of the present invention, there is provided the use of the biomarkers described in the first aspect of the present invention in predicting PD1/PDL1 pathway immune checkpoint inhibitors or PD1/PDL1 pathway immune checkpoint inhibitors combined therapeutic agents for the treatment of tumors in the application of adverse reactions.

In a fourth aspect of the present invention, there is provided a kit for evaluating the response effect of a patient with a tumor after administration of a PD1/PDL1 pathway immune checkpoint inhibitor or a PD1/PDL1 pathway immune checkpoint inhibitor combined therapeutic agent , the kit comprises the biomarkers described in the first aspect of the present invention.

In the fifth aspect of the present invention, there is provided the detection of PD1/PDL1 pathway immune checkpoint inhibitor or PD1/PDL1 pathway immune checkpoint inhibitor combined therapeutic agent in tumors using the kit described in the fourth aspect of the present invention Application in response to effects or adverse reactions.

In the sixth aspect of the present invention, there is provided a method for predicting the response effect or adverse reaction after administration of a PD1/PDL1 pathway immune checkpoint inhibitor or a PD1/PDL1 pathway immune checkpoint inhibitor combined therapeutic agent in a tumor patient, the method include:

1) Extract the patient's peripheral blood mononuclear cells (PBMC), carry out DNA extraction, and carry out quality control;

2) Amplifying and high-throughput sequencing of the CDR3 sequence of the TCRβ chain in the patient's PBMC using a multiplex PCR method;

3) Analyze whether the dominant sequence in the CDR3 sequence of the patient's TCR beta chain contains the biomarker described in the first aspect of the present invention.

In some specific embodiments, the PD1/PDL1 pathway immune checkpoint inhibitor described in the second to sixth aspects of the present invention is an anti-PD1 or anti-PDL1 monoclonal antibody, an anti-PD1 or anti-PDL1 bispecific Antibody, anti-PD1 or anti-PDL1 multispecific antibody, ADC drug targeting PD1 or PDL1 or fusion protein targeting PD1 or PDL1; the therapeutic agent is a small molecule drug, antibody, ADC drug, fusion protein or polypeptide.

More specifically, the above-mentioned anti-PD1 or anti-PDL1 monoclonal antibodies are nivolumab, pembrolizumab, cimilimab, toripalizumab, camrelizumab, tira Libizumab, sintilimab; atezolizumab, avelumab, durvalumab, adebrelimab, pacmilimab, envafolimab; preferably, the anti-PD1 monoclonal antibody is sindi limumab.

In some specific embodiments, the tumor described in the first to sixth aspects of the present invention is Hodgkin's lymphoma; preferably, the tumor is classic Hodgkin's lymphoma.

Invention Definition

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. For the purposes of the present invention, the following terms are defined below.

"Adverse reactions": refers to the occurrence of harmful reactions that are not related to the purpose of treatment during the process of preventing, diagnosing or treating diseases by applying drugs according to normal usage and dosage. Examples include immune-related adverse reactions (irAEs) or other concomitant side effects that occur during treatment.

"Immune checkpoint inhibitors": some drugs developed for corresponding immune checkpoints, whose main function is to block the interaction between tumor cells expressing immune checkpoints and immune cells, thereby blocking the inhibition of immune cells by tumor cells effect.

effect of invention

It can be seen from the technical solution of the present invention that, compared with the prior art, the technical solution of the present invention has the following beneficial effects:

1. The present invention finds for the first time that after a patient with classic Hodgkin lymphoma (cHL) is treated with sintilimab, the change in the abundance of the CDR3 sequence of the TCR-β chain of the patient is related to the efficacy and clinical efficacy of the drug. There is a good correlation between side effects.

2. High-throughput sequencing and statistical analysis were performed on the CDR3 sequence of the TCR-β chain of cHL patients given sintilimab, and the dominant sequence was determined. At the same time, combined with clinical trials and sequence analysis, it was further found that in patients with tumor remission, the CDR3 sequences of the main TCR-β chain were significantly enriched; the dominant sequences tended to coexist in different patients.

3. Among the CDR3 dominant sequences of TCR-β chain of responders, the frequency of "RGG" sequence is extremely high, and the CDR3 sequence of TCR-β chain with the "RGG" sequence can be used as a biomarker for checkpoint immunotherapy thing.

In order to make the above-mentioned and other purposes, features and advantages of the present invention more obvious and easy to understand, preferred embodiments are given below, and are described in detail as follows in conjunction with the accompanying drawings:

Description of drawings

A in Figure 1 shows the experimental steps of Example 1 and the relationship between the efficacy of each patient and the time of administration after administration, wherein CR means complete remission; PR means partial remission; PD means disease progression; irAE means immune related adverse reactions;

B in FIG. 1 shows the sequence number of the total TCR-β chain CDR3 detected by each patient, the sequence number of the TCR-β chain CDR3 sequence obtained by sequencing and the analysis of the consensus sequence among different patients;

C in Figure 1 shows the enrichment abundance of the CDR3 sequences (i.e. dominant sequences) of the highest 3 TCR-β chains in each patient as a percentage of the top 100 sequence abundances, respectively;

D in Figure 1 shows an analysis of the abundance of the CDR3 sequences of the top three TCR-beta chains in each patient (i.e., the dominant sequence) over time, and the presence or absence of "RGG" in the dominant sequence and efficacy The relationship between CR means complete remission; PR means partial remission; irAE means immune-related adverse reactions;

Figure 2 shows the presence in each patient of the CDR3 sequences of up to three TCR-beta chains (ie, predominant sequences) in each patient.

detailed description

The specific embodiments enumerated in the present invention are only used as examples of the present invention, and the present invention is not limited to the specific embodiments described below. For those skilled in the art, any equivalent modifications and substitutions to the embodiments described below are also within the scope of the present invention. Therefore, equivalent changes and modifications made without departing from the spirit and scope of the present invention should be included within the scope of the present invention.

The test materials, test reagents and instruments used in the examples of the present invention are all commercially available.

Example 1:

1. Test materials and test methods

1) Antibody drug: sintilimab

2) Drug source: Innovent Biopharmaceutical (Suzhou) Co., Ltd.

3) Administration mode and administration dose: Sintilimab (ie, Q3W), 200 mg, was administered intravenously every three weeks.

4) Test patients: 4 patients of ORIENT-1 (NCT03114683), numbered p01, p02, p03 and p04, respectively. Among them, the "ORIENT-1" is a multi-center, single-arm, phase II clinical study conducted in China for patients with relapsed or refractory classical Hodgkin lymphoma who have undergone at least second-line systemic chemotherapy. To evaluate the safety and efficacy of sintilimab.

5) Implementation steps: by using a commercial DNA extraction kit (CWE9600 Blood DNA Kit (Auto Plate), CW25345) for the peripheral blood mononuclear cells (PBMC) of 4 patients during sintilimab treatment, and strict DNA extraction from PBMC was performed according to the instructions; the extracted DNA was subjected to quality control, and on this basis, the CDR3 sequence of the TCRβ chain in the PBMC of the patient described by the multiplex PCR method was amplified and high-throughput sequenced.

Wherein, the specific operation steps of the "amplification and high-throughput sequencing" are as follows: TCRβ chain is first amplified by multiplex PCR, and the multiplex PCR contains two stages of PCR1 stage and PCR2 stage, and the primer combination used has been described in the patent (CN105087789A) . The PCR1 stage uses a primer pool pair for the V and J regions of each TCR-β to amplify CDR3, and the PCR2 stage uses a unified primer pool. The specific method of PCR1 stage: template DNA (600ng, 1μl) was added to 25μl of QIAGEN Multiplex PCR master mix, 5μl of Q solution, 1μl of forward primer set library and 1μl of reverse primer set library, and then amplified. The reaction system was formed by using the Multiplex PCR Kit (QIAGEN, Germany). The following PCR program was then performed: 15 min at 95°C followed by 10 cycles of amplification (94°C, 30s denaturation; 60°C, 90s annealing and 72°C, 30s extension). After the final extension at 72°C for 5 min, the system was cooled to 4°C. The product of PCR 1 stage was purified with magnetic beads (Agencourt No. A63882, Beckman, Beverly, MA, USA). All PCR1-stage products were used as templates for PCR2, and PCR2-stage amplification was performed. Add 2 μl primer library, add

Other reagents in the High-Fidelity PCR Kit (New England Biolabs, America) manual, add ultrapure water to a total volume of 50 μl. The following PCR program was then performed: 1 min at 98°C followed by 25 cycles of amplification (98°C, 20s denaturation; 65°C, 30s annealing and 72°C, 30s extension). After the final extension at 72°C for 5 min, the system was cooled to 4°C.

Sequencing: The products of PCR2 stage were gel electrophoresed to recover fragments between 200-350bp, purified by QIAquick Gel Purification Kit (QIAGEN), and Illumina HiSeq3000 was used for paired-end sequencing with a read length of 151bp. The sequencing analysis algorithm was based on the software R (version 3.5.1).

6) Data analysis: ① The enrichment changes of specific CDR3 sequences and the relationship between the changes and the clinical response of patients were analyzed, wherein the “clinical response” refers to the efficacy and immune-related adverse reactions (irAE) or other accompanying side effects . ②We analyzed whether CDR3 sequences coexisted in different patients, and whether these sequences had common sequence features. .

2. Test results

After 3-35 cycles (3 weeks per cycle) of sintilimab, two patients achieved complete response (CR) on p01 and p02, and one patient achieved partial response (CR) on p03 , PR) and showed disease progression (progressiv disease, PD), patient p04 only showed disease progression (progressiv disease, PD). Four patients p01, p02, p03 and p04 all developed irAEs or other concomitant side effects (see A and D in Figure 1 for details). A total of 91,853-225,450 different CDR3 sequences were detected in the four patients, and 88,583-217,043 CDR3 sequences were sequenced respectively (see B in Figure 1 for details). The sum of the abundance of the three most abundant CDR3 sequences in each patient accounts for more than 20% of the abundance of the top 100 sequences, so the three most abundant CDR3 sequences in each patient are defined as the dominant sequences (specifically See C) in Figure 1. We also observed a clear correlation between the enrichment of predominant sequences for each patient and clinical response (see Figure 1, A and D for details).

The specific analysis for 4 patients is as follows (see A, B, C and D in Figure 1 for details):

Patient p01: PR occurred 6 weeks after the initiation of sintilimab treatment, achieved CR after 24 weeks, and then developed irAE - diabetic ketosis at 48 weeks. A total of 217,043 sequences were sequenced against the CDR3 of patient p01. Three dominant sequences were identified: p01-1, p01-2 and p01-3, and the enrichment abundance of these three dominant sequences accounted for 17.1%, 17.0% and 12.0% of the abundance of the top 100 sequences, respectively. Among them, the dominant sequence p01-1: CASSGTSGSTDTQYF (SEQ ID NO: 1) was significantly enriched at the 36th week, and decreased to the basal level at the 48th week. The dominant sequence p01-2: CASSQVSRGGTAEQYF (SEQ ID NO: 2) consistently increased from week 0 and continued to increase, reaching a peak enrichment at week 48. Predominant sequence p01-3: CASTSTLASGANVLTF (SEQ ID NO: 3) was enriched from week 36, and peaked at week 48 at the same time as predominant sequence 2.

Patient p02: achieved PR at 6 weeks after starting sintilimab treatment, achieved CR at 15 weeks, and then developed fibroids at 24 weeks. The fibroid immune response is not clinically defined as an irAE. In studies as other concomitant side effects. A total of 88,583 sequences were sequenced against the CDR3 of patient p02. Three dominant sequences were identified: p02-1, p02-2 and p02-3, and the enrichment abundance of these three dominant sequences accounted for 11.5%, 9.9% and 9.4% of the abundance of the top 100 sequences, respectively. Among them, the dominant sequences p02-1: CASSWEQGARADTQYF (SEQ ID NO: 4) and p02-3: CASSLGRGGQGSGYTF (SEQ ID NO: 6) began to enrich after PR appeared, and the dominant sequences p02-2: CASSLVLSTSYEQYF (SEQ ID NO: 5) ) began to enrich after the appearance of CR, and the three dominant sequences all reached their peaks at the 24th week.

Patient p03: achieved tumor remission after sintilimab treatment, developed irAE pneumonitis at week 6 and had to discontinue treatment. A total of 133,368 sequences were obtained by CDR3 sequencing. Three dominant sequences were identified: p03-1, p03-2 and p03-3, and the enrichment abundance of these three dominant sequences accounted for 72.8%, 3.9% and 3.7% of the abundance of the top 100 sequences, respectively. Among them, the dominant sequence p03-1: CASSIAGGSYEQYF (SEQ ID NO: 7) accounted for 65% of the entire TCR pool at baseline. The dominant sequences p03-2: CASRGGSYEQYF (SEQ ID NO: 8) and p03-3: CASSNRGGNEQFF (SEQ ID NO: 9) reached peak enrichment at week 6. Despite irAEs, p03 achieved PR from week 14 to week 36 after discontinuation.

Patient p04: No tumor remission during sintilimab treatment. Hepatic dysfunction of the irAE occurred at week 36 and treatment had to be discontinued. A total of 99,290 sequences were sequenced for CDR3. Three dominant sequences p04-1, p04-2 and p04-3 were identified, and the enrichment abundance of these three dominant sequences accounted for 16.7%, 4.3% and 2.9% of the abundance of the top 100 sequences, respectively. Among them, the dominant sequence p04-1: CASSHNRGNEEKLFF (SEQ ID NO: 10) was significantly enriched at week 36, the dominant sequence p04-2: CASSPSAGRFGEQYF (SEQ ID NO: 11) and p04-3: CASSYKLTGVANYGYTF (SEQ ID NO: 12) No significant enrichment occurred at 36 weeks.

3. Conclusion

Based on the above test results, the following conclusions can be drawn:

Conclusion 1: When patients achieved tumor remission, the major TCR-β chain CDR3 sequences were significantly enriched, and the highly variable TCR-β chain CDR3 sequences were also closely associated with irAEs or other concomitant side effects. Specifically,

1) The dominant CDR3 sequences tend to co-exist in different patients.

Of the pool of CDR3 sequences detectable in each patient, 3.6%-7.1% of the sequences were shared by two or more individuals, and a total of 191 sequences were shared by all 4 patients (see Figure 1 for details). of B). It is worth noting that in the above analysis, the top three predominant sequences (that is, 9 sequences from p01-1 to p03-3) of the p01, p02 and p03 patients with curative effect were all shared by at least two patients; the right half of Figure 2 The light gray parts corresponding to parts p01-p04 (that is, the parts indicated by "yes" in the legend of Fig. 2) represent that they can be detected in the corresponding 4 patients. In particular, 5 of the 6 dominant sequences (i.e., p01-1, p01-2, p02-1, p02-2, and p02-3) in p01 and p02 of the 2 patients with CR response were composed of 4 Patients shared, except that the above-mentioned 5 dominant sequences were not defined as dominant sequences in p03 and p04 (see the left half of Figure 2 for details).

2) "RGG" also has some common sequence features.

Although the CDR3 sequences of the dominant TCR-beta chain were not identical in treatment-responsive patients, at least one RGG-containing CDR3 sequence was present in these patients. Since the CDR3 sequences contribute significantly to TCR binding to antigen, the presence of a common RGG indicates binding to the same antigen.

Since the CDR3 sequences of TCR-β chains shared between different patients may reflect the similarity of TCR recognition antigens ^[10-11] , the GLIPH clustering algorithm ^{[10] was used} (this algorithm can identify the same specificity based on sequence similarity. Clustering TCR sequences to better analyze the large number of TCR sequences that have been identified, the algorithm can also predict the HLA restriction of these TCR clusters based on the subject's genotype, thereby helping to explore new antigens) Analysis of these sequences in Amino acid similarity. It was found that sequences containing "RGG" appeared most frequently among the 12 dominant sequences in 4 patients (p<0.0001, see Figure 2 for details). More importantly, this RGG sequence was only present in responders (ie present in patients p01, p02 and p03) and was absent in non-responding patient p04.

Conclusion 2: The common TCR-β chain-CDR3 sequence may recognize tumor antigens associated with cHL. Some of these common sequences are more reactive and can expand rapidly after PD-1 blockade therapy. Once TCRs with these "highly occurring and co-existing" dominant sequences (9 sequences p01-1 to p03-3 as shown in Figure 2) predominate in a given T cell pool, their expansion may would lead to a better antitumor response after immunotherapy. In addition, baseline TCR abundance and expansion of dominant sequences were also associated with the incidence of clinical irAEs or other concomitant side effects.

Specifically, in patient p01: the occurrence of diabetic ketosis irAEs coincided with the occurrence of the highest abundance of the predominant sequences p01-2 and p01-3; in patient p02: the occurrence of fibroid complications coincided with the predominant sequence p02-1 , the highest abundance of p02-2 and p02-3 occurred at the same time; in patient p03: the dominant sequence p03-1 was extremely abundant, which may be related to the occurrence of pneumonia soon after PD-1 treatment. In addition, it is also In the occurrence of pneumonia, p02-2 and p02-3 reached the highest abundance; in patient p04: the occurrence of liver dysfunction irAE coincided with the occurrence of the highest abundance of the predominant sequence p04-1.

Conclusion 3: Enrichment analysis of the CDR3 sequences of TCR-β chains in patients with administration, especially the analysis of the dominant sequences with the "RGG" feature, is helpful for monitoring and predicting the response and irAEs after checkpoint immunotherapy .

Patient p01, patient p02, and patient p03 were patients with clinical response to treatment, and all of their three dominant TCR-β chain CDR3 sequences had one or two RGG-containing sequences, and all of them were in RGG-containing sequences. irAEs or other immune responses occur when the highest abundance of enrichment is reached. However, in the CDR3 sequence of the dominant TCR-β chain in p04 of the only patient with progressive disease (corresponding patient with no response), no RGG sequence was found.

references:

1. Shi, Y., et al., Safety and activity of sintilimab in patients with relapsed or refractory classic Hodgkin lymphoma (ORIENT-1): a multicentre, single-arm, phase 2 trial. Lancet Haematol, 2019.6(1): p.e12-e19.

2. Armand, P., et al., Nivolumab for Relapsed/Refractory Classic Hodgkin Lymphoma After Failure of Autologous Hematopoietic Cell Transplantation: Extended Follow-Up of the Multicohort Single-Arm Phase II CheckMate 205 Trial.J Clin Oncol, 2018.36(14 ): p.1428-1439.

3. Chen, R., et al., Pembrolizumab in relapsed or refractory Hodgkin lymphoma: 2-year follow-up of KEYNOTE-087. Blood, 2019.134(14): p.1144-1153.

4. Meti, N., K. Esfahani, and N.A. Johnson, The Role of Immune Checkpoint Inhibitors in Classical Hodgkin Lymphoma. Cancers (Basel), 2018.10(6).

5. De Goycoechea, D., et al., Immune Checkpoint Inhibition in Classical Hodgkin Lymphoma: From Early Achievements towards New Perspectives. J Oncol, 2019.2019: p.9513701.

6. Berner, F., et al., Association of Checkpoint Inhibitor-Induced Toxic Effects With Shared Cancer and Tissue Antigens in Non-Small Cell Lung Cancer. JAMA Oncol, 2019.5(7):p.1043-1047.

7. Pietsch, E.C., et al., Anti-leukemic activity and tolerability of anti-human CD47 monoclonal antibodies. Blood Cancer J, 2017.7(2):p.e536.

8. Abbas, A.K., et al., Cellular and molecular immunology. Ninth edition.ed. 2018, Philadelphia, PA: Elsevier.x, 565pages.

9. Cabaniols, J.P., et al., Most alpha/beta T cell receptor diversity is due to terminal deoxynucleotidyl transferase. J Exp Med, 2001.194(9):p.1385-90.

10. Glanville, J., et al., Identifying specificity groups in the T cell receptor repertoire. Nature, 2017.547(7661): p.94-98.

11. Madi, A., et al., T-cell receptor repertoires share a restricted set of public and abundant CDR3 sequences that are associated with self-related immunity. Genome Res, 2014.24(10): p.1603-12.

Claims (10)

1. A set of biomarkers is characterized in that, the biomarker is a consensus amino acid sequence in the CDR3 sequence of T cell receptor (TCR) beta chain; preferably, the amino acid sequence of the biomarker includes RGG.
2. The biomarker according to claim 1, wherein the biomarkers respectively comprise the amino acid sequence shown in any one of SEQ ID NO: 1 to SEQ ID NO: 9; preferably, the biomarkers The biomarkers respectively comprise the amino acid sequences shown in any one of SEQ ID NO: 1-2, SEQ ID NO: 4-6 or SEQ ID NO: 7; more preferably, the biomarkers respectively comprise SEQ ID NO: 2 or the amino acid sequence shown in SEQ ID NO:6.
3. The application of the biomarkers described in claim 1 or 2 in the preparation of a kit for predicting the response effect of PD1/PDL1 pathway immune checkpoint inhibitor or PD1/PDL1 pathway immune checkpoint inhibitor combined therapeutic agent.
4. Application of the biomarkers described in claim 1 or 2 in predicting adverse reactions of PD1/PDL1 pathway immune checkpoint inhibitors or PD1/PDL1 pathway immune checkpoint inhibitors combined therapeutic agents in the treatment of tumors.
5. A kit for evaluating the response effect of PD1/PDL1 pathway immune checkpoint inhibitor or PD1/PDL1 pathway immune checkpoint inhibitor combined therapeutic agent in a patient with tumor, the kit comprising the method as claimed in the claim The biomarkers described in 1 or 2.
6. The application of the kit according to claim 5 in detecting the response effect or adverse reaction of PD1/PDL1 pathway immune checkpoint inhibitor or PD1/PDL1 pathway immune checkpoint inhibitor combined therapeutic agent in tumor.
7. A method for predicting the response effect or adverse reaction after administration of a PD1/PDL1 pathway immune checkpoint inhibitor or a PD1/PDL1 pathway immune checkpoint inhibitor combined therapeutic agent in a tumor patient, the method comprising:

   1) Extract the patient's peripheral blood mononuclear cells (PBMC) for DNA extraction and quality control;

   2) Amplifying and high-throughput sequencing of the CDR3 sequence of the TCRβ chain in the patient's PBMC using a multiplex PCR method;

   3) Analyze whether the predominant sequence in the CDR3 sequence of the patient's TCR beta chain contains the biomarker according to claim 1 or 2.
8. The application according to claim 3, 4 or 6, the kit according to claim 5, and the method according to claim 7, wherein the PD1/PDL1 pathway immune checkpoint inhibitor is anti-PD1 or anti-PD1 Monoclonal antibody against PDL1, bispecific antibody against PD1 or PDL1, multispecific antibody against PD1 or PDL1, ADC drug targeting PD1 or PDL1, or fusion protein targeting PD1 or PDL1; the therapeutic agent It is a small molecule drug, antibody, ADC drug, fusion protein or polypeptide.
9. The application, kit or method according to claim 8, wherein the anti-PD1 or anti-PDL1 monoclonal antibody is nivolumab, pembrolizumab, similimab, toripril mAb, camrelizumab, tislelizumab, sintilimab; atezolizumab, avelumab, durvalumab, adebrelimab, pacmilimab, envafolimab; preferred , the anti-PD1 monoclonal antibody is sintilimab.
10. The marker according to claim 1, the application according to claim 3, 4 or 6, the kit according to claim 5, and the method according to claim 7, wherein the tumor is Hodgkin lymphoma; preferably, the tumor is classic Hodgkin lymphoma.

Images