Gene combination for screening different modes of alloHCT pre-donors and predicting aGVHD risk after transplantation and application of gene combination

Abstract

The invention discloses a gene combination for screening different modes of alloHCT pre-donors and predicting aGVHD risk after transplantation and application of the gene combination. According to the invention, a gene combination with prediction and diagnosis values for aGVHD after HLA haplotype congruence and HLA full congruence alloHCT is found through a whole genome sequencing and target gene capture sequencing method. On one hand, comprehensive judgment and evaluation are carried out by detecting gene combination mutation conditions of donors and recipients, selection of donors before alloHCT is optimized, and the survival rate and life quality of patients are improved; on the other hand, the possibility of aGVHD of alloHCT recipients in different modes can be predicted by detecting gene combination mutation conditions of the donor and the recipients, and a basis is provided for potential aGVHD diagnosis; and thirdly, a new genetic basis is provided for revealing the pathogenesis of the aGVHD, and a new drug target is provided for treating the aGVHD.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to a gene combination and application thereof for pre-donor screening of alloHCT in different modes and post-transplant aGVHD risk prediction. Background Art

[0002] Allogeneic hematopoietic cell transplantation (alloHCT) can effectively treat more than 70 diseases, including leukemia, genetic diseases, and autoimmune diseases. Acute graft-versus-host disease (aGVHD) is a common fatal complication after alloHCT, which seriously affects the prognosis and survival of patients. For a long time, whether the HLA genotype between the donor and the recipient is matched has been considered to be the decisive factor affecting the occurrence of aGVHD. Selecting a donor who is completely HLA-matched with the recipient was once the "gold standard" in the field of alloHCT. After decades of development and improvement, the HLA haplotype-matched alloHCT regimen based on antithymocyte globulin (ATG) and granulocyte colony-stimulating factor (G-CSF)-induced immune tolerance originally developed by the Institute of Hematology of Peking University has achieved clinical efficacy comparable to that of HLA-matched transplantation, becoming the most important alloHCT model in my country and widely used in countries around the world.

[0003] The great success of the HLA haplotype-matched transplantation program means that the classic immunological barrier of HLA incompatibility has been overcome in clinical practice. However, aGVHD will occur after both HLA haplotype-matched and HLA fully matched transplantation, which highly suggests that there are other genetic factors that affect the occurrence of aGVHD. On the other hand, the successful application of HLA haplotype-matched transplantation has greatly solved the problem of lack of donor sources, making the patient's parents, children, siblings and collateral relatives potential candidate donors. When a patient has multiple potential donors, how to choose the most suitable donor is of great significance for reducing the risk of aGVHD and improving the patient's overall survival rate. Summary of the invention

[0004] The object of the present invention is to provide a gene combination and its application for donor screening before HLA haplotype matching and HLA fully matching allogeneic hematopoietic cell transplantation (alloHCT) and risk prediction of acute graft-versus-host disease (aGVHD) after alloHCT.

[0005] In order to achieve the above-mentioned object, the present invention first provides a new use of a substance for detecting whether a gene pair in a donor and a recipient has mutated.

[0006] The present invention provides the use of a substance for detecting whether a gene pair in a donor and a recipient is mutated in any one of the following A1)-A4):

[0007] A1) Preparation of products for screening or assisting in the screening of donors prior to allogeneic hematopoietic cell transplantation;

[0008] A2) Preparation of products for matching or assisting matching between donors and recipients before allogeneic hematopoietic cell transplantation (matching or assisting matching between donors and recipients before alloHCT);

[0009] A3) preparing products for predicting or assisting in predicting the risk of acute graft-versus-host disease in patients after allogeneic hematopoietic cell transplantation;

[0010] A4) Prepare products for diagnosing or assisting in the diagnosis of acute graft-versus-host disease in patients after allogeneic hematopoietic cell transplantation.

[0011] The present invention also provides the use of a substance for detecting whether a gene pair in a donor and a recipient has mutated in any of the following B1)-B4):

[0012] B1) Screening or assisting in the selection of donors before allogeneic hematopoietic cell transplantation;

[0013] B2) matching or auxiliary matching between donors and recipients before allogeneic hematopoietic cell transplantation (matching or auxiliary matching between donors and recipients before alloHCT);

[0014] B3) predict or assist in predicting the risk of acute graft-versus-host disease in patients after allogeneic hematopoietic cell transplantation;

[0015] B4) Diagnose or assist in the diagnosis of acute graft-versus-host disease in patients after allogeneic hematopoietic cell transplantation.

[0016] In order to achieve the above object, the present invention also provides a kit having any one of the following functions a1)-a4), which comprises a substance for detecting whether a gene pair in a donor and a recipient has mutated;

[0017] a1) Screening or assisting in the selection of donors before allogeneic hematopoietic cell transplantation;

[0018] a2) Matching or auxiliary matching between donors and recipients before hematopoietic cell transplantation (matching or auxiliary matching between donors and recipients before alloHCT);

[0019] a3) To predict or assist in predicting the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation;

[0020] a4) To diagnose or assist in the diagnosis of acute graft-versus-host disease in patients after hematopoietic cell transplantation.

[0021] In any of the above applications or kits, the gene pair is at least one of the following gene pairs: ADAMTS7-APOL4, AGAP3-ACR, ANAPC1-ACR, CARD8-SIRPB1, DCAF15-ACR, DENND3-ACR, FRMD5-MYCT1, HAPLN3-ACR, HRNR-ACR, MAGEE2-ACR, MYCT1-KRTAP10-1, MYCT1-OAS1, NUDT1-ACR, OAS1-ACR, PABPC3-ACR, SIRPA-ACR, SLC35G6- ACR, TLR8-ACR, ZFPM1-ACR, ACTN2-OBSCN, CDH15-FRMD5, CDH15-GPR161, CDH15-OBSCN, GNRH2-FRMD5, GNRH2-KRT24, GNRH2-MAGEE2, G NRH2-OBSCN, GPSM1-OBSCN, HAPLN3-OBSCN, LCN10-CD33, MS4A12-ACTN2, MS4A12-ZNF211, RBMX-CD33, SIRPB1-ACTN2, SIRPB1-OBSCN.

[0022] In any of the above applications or kits, the two genes before and after the "-" in each gene pair correspond to the donor and the recipient, respectively, that is, the gene before the "-" in each gene pair is recorded as the first gene, and the gene after the "-" is recorded as the second gene. The detection of whether a gene pair in the donor-recipient is mutated refers to detecting whether a mutation occurs in the first gene in the donor and the second gene in the recipient.

[0023] In any of the above applications or kits, the substance for detecting whether a gene pair in a donor and a recipient has mutated may be a substance for detecting whether a gene pair in the peripheral blood of a donor and a recipient has mutated.

[0024] Furthermore, the substance for detecting whether a gene pair in the peripheral blood of a donor and a recipient has mutated may be a reagent and / or an instrument for detecting whether a gene pair in the peripheral blood of a donor and a recipient has mutated.

[0025] The reagents and / or instruments include but are not limited to reagents for extracting genomic DNA from human peripheral blood cells, and reagents and / or instruments required for detecting whether gene pairs in peripheral blood are mutated using various conventional methods in the prior art (such as DNA sequencing, restriction fragment length polymorphism, single-stranded conformation polymorphism, allele-specific oligonucleotide hybridization, etc.).

[0026] Furthermore, the reagents and / or instruments required for detecting whether a gene pair in peripheral blood is mutated using DNA sequencing (such as high-throughput sequencing or first-generation sequencing, etc.) include various primers, probes and sequencers used in sequencing.

[0027] In a specific embodiment of the present invention, the DNA sequencing method may specifically include the following steps:

[0028] 1. Draw venous blood from donors and recipients before alloHCT and extract genomic DNA;

[0029] 2. Use the enzyme digestion method to randomly fragment 200ng genomic DNA into an average length of 150-200bp, then repair and fill the ends and add A bases at the 3' end to form sticky ends, then connect with the adapter containing the specific barcode sequence, and then use magnetic beads to screen to remove incomplete connection products and adapter self-connection products, and finally use universal primers complementary to the adapter sequence for PCR amplification to form a sequencing library;

[0030] 3. Use The pool probes were hybridized in the hybridization buffer, and then the hybridized target fragments were captured using magnetic beads and separated and purified. The captured DNA fragments were amplified by PCR and the quality of the captured library was checked (including Qubit concentration determination and 2100 Bioanalyzer fragment length determination). Finally, PE150 high-throughput sequencing was performed on the NovaSeq6000 sequencer according to Illumina's standard process.

[0031] 4. Use sequencing profiles for genotyping.

[0032] In any of the above applications or kits, the kit further comprises a data processing device, wherein the data processing device has a built-in module, and the module has the following functions: predicting the risk of acute graft-versus-host disease in patients after alloHCT or diagnosing whether acute graft-versus-host disease occurs in patients after alloHCT based on whether the above gene pair in the peripheral blood of the donor and the recipient is mutated.

[0033] In practical applications, any one of the 35 gene pairs mentioned above can be used to predict the risk of acute graft-versus-host disease in patients after alloHCT, to diagnose whether acute graft-versus-host disease occurs in patients after alloHCT, or to screen donors before alloHCT.

[0034] Specifically,

[0035] The method for predicting the risk of acute graft-versus-host disease in patients after alloHCT based on whether the ADAMTS7-APOL4 gene pair is mutated in the donor and recipient is as follows: the risk of acute graft-versus-host disease in patients after alloHCT with mutations in the donor ADAMTS7 gene and mutations in the recipient APOL4 gene is lower than that in patients after alloHCT in whom at least one of the donor ADAMTS gene and the recipient APOL4 gene is not mutated.

[0036] The method for diagnosing whether acute graft-versus-host disease occurs in patients after alloHCT based on whether the ADAMTS7-APOL4 gene pair in the donor and the recipient is mutated is as follows: if the donor ADAMTS7 gene is mutated and the recipient APOL4 gene is mutated, the patient develops or is a candidate for developing acute graft-versus-host disease after alloHCT; if at least one of the donor ADAMTS7 gene and the recipient APOL4 gene is not mutated, the patient does not develop or is a candidate for not developing acute graft-versus-host disease after alloHCT.

[0037] The method for predicting the risk of acute graft-versus-host disease in patients after alloHCT based on whether the AGAP3-ACR gene pair in the donor and recipient is mutated is as follows: the risk of acute graft-versus-host disease in patients after alloHCT with both a mutation in the donor AGAP3 gene and a mutation in the recipient ACR gene is lower than that in patients after alloHCT in whom at least one of the donor AGAP3 gene and the recipient ACR gene is not mutated.

[0038] The method for diagnosing whether acute graft-versus-host disease occurs in patients after alloHCT based on whether the AGAP3-ACR gene pair in the donor and the recipient is mutated is as follows: if the donor AGAP3 gene is mutated and the recipient ACR gene is mutated, the patient does not develop or is a candidate for not developing acute graft-versus-host disease after alloHCT; if at least one of the donor AGAP3 gene and the recipient ACR gene is not mutated, the patient develops or is a candidate for developing acute graft-versus-host disease after alloHCT.

[0039] The method for predicting the risk of acute graft-versus-host disease in patients after alloHCT based on whether the ANAPC1-ACR gene pair in the donor and recipient is mutated is as follows: the risk of acute graft-versus-host disease in patients after alloHCT with mutations in the donor ANAPC1 gene and mutations in the recipient ACR gene is lower than that in patients after alloHCT in whom at least one of the donor ANAPC1 gene and the recipient ACR gene is not mutated.

[0040] The method for diagnosing whether acute graft-versus-host disease occurs in patients after alloHCT based on whether the ANAPC1-ACR gene pair in the donor and the recipient is mutated is as follows: if the donor ANAPC1 gene is mutated and the recipient ACTN2 gene is mutated, the patient does not develop or is a candidate for not developing acute graft-versus-host disease after alloHCT; if at least one of the donor ANAPC1 gene and the recipient ACR gene is not mutated, the patient develops or is a candidate for developing acute graft-versus-host disease after alloHCT.

[0041] The method for predicting the risk of acute graft-versus-host disease in patients after alloHCT based on whether the CARD8-SIRPB1 gene pair is mutated in the donor and recipient is as follows: the risk of acute graft-versus-host disease in patients after alloHCT with mutations in the donor CARD8 gene and mutations in the recipient SIRPB1 gene is lower than that in patients after alloHCT in whom at least one of the donor CARD8 gene and the recipient SIRPB1 gene is not mutated.

[0042] The method for diagnosing whether acute graft-versus-host disease occurs in patients after alloHCT based on whether the CARD8-SIRPB1 gene pair in the donor and the recipient is mutated is as follows: if the donor's CARD8 gene is mutated and the recipient's SIRPB1 gene is mutated, the patient does not develop or is a candidate for not developing acute graft-versus-host disease after alloHCT; if at least one of the donor's CARD8 gene and the recipient's SIRPB1 gene is not mutated, the patient develops or is a candidate for developing acute graft-versus-host disease after alloHCT.

[0043] The method for predicting the risk of acute graft-versus-host disease in patients after alloHCT based on whether the DCAF15-ACR gene pair is mutated in the donor and recipient is as follows: the risk of acute graft-versus-host disease in patients after alloHCT with mutations in the donor DCAF15 gene and mutations in the recipient ACR gene is lower than that in patients after alloHCT in whom at least one of the donor DCAF15 gene and the recipient ACR gene is not mutated.

[0044] The method for diagnosing whether acute graft-versus-host disease occurs in patients after alloHCT based on whether the DCAF15-ACR gene pair in the donor and the recipient is mutated is as follows: if the donor DCAF15 gene is mutated and the recipient ACR gene is mutated, the patient does not develop or is a candidate for not developing acute graft-versus-host disease after alloHCT; if at least one of the donor DCAF15 gene and the recipient ACR gene is not mutated, the patient develops or is a candidate for developing acute graft-versus-host disease after alloHCT.

[0045] The method for predicting the risk of acute graft-versus-host disease in patients after alloHCT based on whether the DENND3-ACR gene pair in the donor and recipient is mutated is as follows: the risk of acute graft-versus-host disease in patients after alloHCT with mutations in the donor DENND3 gene and mutations in the recipient ACR gene is lower than that in patients after alloHCT in whom at least one of the donor DENND3 gene and the recipient ACR gene is not mutated.

[0046] The method for diagnosing whether acute graft-versus-host disease occurs in patients after alloHCT based on whether the DENND3-ACR gene pair in the donor and the recipient is mutated is as follows: if the donor DENND3 gene is mutated and the recipient ACR gene is mutated, the patient does not develop or is a candidate for not developing acute graft-versus-host disease after alloHCT; if at least one of the donor DENND3 gene and the recipient ACR gene is not mutated, the patient develops or is a candidate for developing acute graft-versus-host disease after alloHCT.

[0047] The method for predicting the risk of acute graft-versus-host disease in patients after alloHCT based on whether the FRMD5-MYCT1 gene pair is mutated in the donor and recipient is as follows: the risk of acute graft-versus-host disease in patients after alloHCT with both a mutation in the donor FRMD5 gene and a mutation in the recipient MYCT1 gene is lower than that in patients after alloHCT in whom at least one of the donor FRMD5 gene and the recipient MYCT1 gene is not mutated.

[0048] The method for diagnosing whether acute graft-versus-host disease occurs in patients after alloHCT based on whether the FRMD5-MYCT1 gene pair in the donor and the recipient is mutated is as follows: if the donor's FRMD5 gene is mutated and the recipient's MYCT1 gene is mutated, the patient does not develop or is a candidate for not developing acute graft-versus-host disease after alloHCT; if at least one of the donor's FRMD5 gene and the recipient's MYCT1 gene is not mutated, the patient develops or is a candidate for developing acute graft-versus-host disease after alloHCT.

[0049] The method for predicting the risk of acute graft-versus-host disease in patients after alloHCT based on whether the HAPLN3-ACR gene pair is mutated in the donor and recipient is as follows: the risk of acute graft-versus-host disease in patients after alloHCT with mutations in the donor HAPLN3 gene and mutations in the recipient ACR gene is lower than that in patients after alloHCT in whom at least one of the donor HAPLN3 gene and the recipient ACR gene is not mutated.

[0050] The method for diagnosing whether acute graft-versus-host disease occurs in patients after alloHCT based on whether the HAPLN3-ACR gene pair in the donor and the recipient is mutated is as follows: if the donor HAPLN3 gene is mutated and the recipient ACR gene is mutated, the patient does not develop or is a candidate for not developing acute graft-versus-host disease after alloHCT; if at least one of the donor HAPLN3 gene and the recipient ACR gene is not mutated, the patient develops or is a candidate for developing acute graft-versus-host disease after alloHCT.

[0051] The method for predicting the risk of acute graft-versus-host disease in patients after alloHCT based on whether the HRNR-ACR gene pair in the donor and recipient is mutated is as follows: the risk of acute graft-versus-host disease in patients after alloHCT with mutations in the donor HRNR gene and mutations in the recipient ACR gene is lower than that in patients after alloHCT in whom at least one of the donor HRNR gene and the recipient ACR gene is not mutated.

[0052] The method for diagnosing whether acute graft-versus-host disease occurs in patients after alloHCT based on whether the HRNR-ACR gene pair in the donor and the recipient is mutated is as follows: if the donor HRNR gene is mutated and the recipient ACR gene is mutated, the patient does not develop or is a candidate for not developing acute graft-versus-host disease after alloHCT; if at least one of the donor HRNR gene and the recipient ACR gene is not mutated, the patient develops or is a candidate for developing acute graft-versus-host disease after alloHCT.

[0053] The method for predicting the risk of acute graft-versus-host disease in patients after alloHCT based on whether the MAGEE2-ACR gene pair in the donor and recipient is mutated is as follows: the risk of acute graft-versus-host disease in patients after alloHCT with mutations in the donor MAGEE2 gene and mutations in the recipient ACR gene is lower than that in patients after alloHCT in whom at least one of the donor MAGEE2 gene and the recipient ACR gene is not mutated.

[0054] The method for diagnosing whether acute graft-versus-host disease occurs in patients after alloHCT based on whether the MAGEE2-ACR gene pair in the donor and the recipient is mutated is as follows: if the donor MAGEE2 gene is mutated and the recipient ACR gene is mutated, the patient does not develop or is a candidate for not developing acute graft-versus-host disease after alloHCT; if at least one of the donor MAGEE2 gene and the recipient ACR gene is not mutated, the patient develops or is a candidate for developing acute graft-versus-host disease after alloHCT.

[0055] The method for predicting the risk of acute graft-versus-host disease in patients after alloHCT based on whether the MYCT1-KRTAP10-1 gene pair in the donor and recipient is mutated is as follows: the risk of acute graft-versus-host disease in patients after alloHCT with both the donor MYCT1 gene mutation and the recipient KRTAP10-1 gene mutation is lower than that in patients after alloHCT in whom at least one of the donor MYCT1 gene and the recipient KRTAP10-1 gene is not mutated.

[0056] The method for diagnosing whether acute graft-versus-host disease occurs in patients after alloHCT based on whether the MYCT1-KRTAP10-1 gene pair in the donor and the recipient is mutated is as follows: if the donor MYCT1 gene is mutated and the recipient KRTAP10-1 gene is mutated, the patient does not develop or is a candidate for not developing acute graft-versus-host disease after alloHCT; if at least one of the donor MYCT1 gene and the recipient KRTAP10-1 gene is not mutated, the patient develops or is a candidate for developing acute graft-versus-host disease after alloHCT.

[0057] The method for predicting the risk of acute graft-versus-host disease in patients after alloHCT based on whether the MYCT1-OAS1 gene pair in the donor and recipient is mutated is as follows: the risk of acute graft-versus-host disease in patients after alloHCT with both a mutation in the donor MYCT1 gene and a mutation in the recipient OAS1 gene is lower than that in patients after alloHCT in whom at least one of the donor MYCT1 gene and the recipient OAS1 gene is not mutated.

[0058] The method for diagnosing whether acute graft-versus-host disease occurs in patients after alloHCT based on whether the MYCT1-OAS1 gene pair in the donor and the recipient is mutated is as follows: if the donor's MYCT1 gene is mutated and the recipient's OAS1 gene is mutated, the patient does not develop or is a candidate for not developing acute graft-versus-host disease after alloHCT; if at least one of the donor's MYCT1 gene and the recipient's OAS1 gene is not mutated, the patient develops or is a candidate for developing acute graft-versus-host disease after alloHCT.

[0059] The method for predicting the risk of acute graft-versus-host disease in patients after alloHCT based on whether the NUDT1-ACR gene pair in the donor and recipient is mutated is as follows: the risk of acute graft-versus-host disease in patients after alloHCT with mutations in the donor NUDT1 gene and mutations in the recipient ACR gene is lower than that in patients after alloHCT in whom at least one of the donor NUDT1 gene and recipient ACR gene is not mutated.

[0060] The method for diagnosing whether acute graft-versus-host disease occurs in patients after alloHCT based on whether the NUDT1-ACR gene pair in the donor and the recipient is mutated is as follows: if the donor NUDT1 gene is mutated and the recipient ACR gene is mutated, the patient does not develop or is a candidate for not developing acute graft-versus-host disease after alloHCT; if at least one of the donor NUDT1 gene and the recipient ACR gene is not mutated, the patient develops or is a candidate for developing acute graft-versus-host disease after alloHCT.

[0061] The method for predicting the risk of acute graft-versus-host disease in patients after alloHCT based on whether the OAS1-ACR gene pair in the donor and recipient is mutated is as follows: the risk of acute graft-versus-host disease in patients after alloHCT with both a mutation in the donor OAS1 gene and a mutation in the recipient ACR gene is lower than that in patients after alloHCT in whom at least one of the donor OAS1 gene and the recipient ACR gene is not mutated.

[0062] The method for diagnosing whether acute graft-versus-host disease occurs in patients after alloHCT based on whether the OAS1-ACR gene pair in the donor and the recipient is mutated is as follows: if the donor OAS1 gene is mutated and the recipient ACR gene is mutated, the patient does not develop or is a candidate for not developing acute graft-versus-host disease after alloHCT; if at least one of the donor OAS1 gene and the recipient ACR gene is not mutated, the patient develops or is a candidate for developing acute graft-versus-host disease after alloHCT.

[0063] The method for predicting the risk of acute graft-versus-host disease in patients after alloHCT based on whether the PABPC3-ACR gene pair in the donor and recipient is mutated is as follows: the risk of acute graft-versus-host disease in patients after alloHCT with mutations in the donor PABPC3 gene and mutations in the recipient ACR gene is lower than that in patients after alloHCT in whom at least one of the donor PABPC3 gene and the recipient ACR gene is not mutated.

[0064] The method for diagnosing whether acute graft-versus-host disease occurs in patients after alloHCT based on whether the PABPC3-ACR gene pair in the donor and the recipient is mutated is as follows: if the donor's PABPC3 gene is mutated and the recipient's ACR gene is mutated, the patient does not develop or is a candidate for not developing acute graft-versus-host disease after alloHCT; if at least one of the donor's PABPC3 gene and the recipient's ACR gene is not mutated, the patient develops or is a candidate for developing acute graft-versus-host disease after alloHCT.

[0065] The method for predicting the risk of acute graft-versus-host disease in patients after alloHCT based on whether the SIRPA-ACR gene pair in the donor and recipient is mutated is as follows: the risk of acute graft-versus-host disease in patients after alloHCT with mutations in the donor SIRPA gene and mutations in the recipient ACR gene is lower than that in patients after alloHCT in whom at least one of the donor SIRPA gene and the recipient ACR gene is not mutated.

[0066] The method for diagnosing whether acute graft-versus-host disease occurs in patients after alloHCT based on whether the SIRPA-ACR gene pair in the donor and the recipient is mutated is as follows: if the donor SIRPA gene is mutated and the recipient ACR gene is mutated, the patient does not develop or is a candidate for not developing acute graft-versus-host disease after alloHCT; if at least one of the donor SIRPA gene and the recipient ACR gene is not mutated, the patient develops or is a candidate for developing acute graft-versus-host disease after alloHCT.

[0067] The method for predicting the risk of acute graft-versus-host disease in patients after alloHCT based on whether the SLC35G6-ACR gene pair is mutated in the donor and recipient is as follows: the risk of acute graft-versus-host disease in patients after alloHCT with mutations in the donor SLC35G6 gene and mutations in the recipient ACR gene is lower than that in patients after alloHCT in whom at least one of the donor SLC35G6 gene and the recipient ACR gene is not mutated.

[0068] The method for diagnosing whether acute graft-versus-host disease occurs in patients after alloHCT based on whether the SLC35G6-ACR gene pair in the donor and the recipient is mutated is as follows: if the donor SLC35G6 gene is mutated and the recipient ACR gene is mutated, the patient does not develop or is a candidate for not developing acute graft-versus-host disease after alloHCT; if at least one of the donor SLC35G6 gene and the recipient ACR gene is not mutated, the patient develops or is a candidate for developing acute graft-versus-host disease after alloHCT.

[0069] The method for predicting the risk of acute graft-versus-host disease in patients after alloHCT based on whether the TLR8-ACR gene pair in the donor and recipient is mutated is as follows: the risk of acute graft-versus-host disease in patients after alloHCT with both a mutation in the donor TLR8 gene and a mutation in the recipient ACR gene is lower than that in patients after alloHCT in whom at least one of the donor TLR8 gene and the recipient ACR gene is not mutated.

[0070] The method for diagnosing whether acute graft-versus-host disease occurs in patients after alloHCT based on whether the TLR8-ACR gene pair in the donor and the recipient is mutated is as follows: if the donor TLR8 gene is mutated and the recipient ACR gene is mutated, the patient does not develop or is a candidate for not developing acute graft-versus-host disease after alloHCT; if at least one of the donor TLR8 gene and the recipient ACR gene is not mutated, the patient develops or is a candidate for developing acute graft-versus-host disease after alloHCT.

[0071] The method for predicting the risk of acute graft-versus-host disease in patients after alloHCT based on whether the ZFPM1-ACR gene pair in the donor and recipient is mutated is as follows: the risk of acute graft-versus-host disease in patients after alloHCT with mutations in the donor ZFPM1 gene and mutations in the recipient ACR gene is lower than that in patients after alloHCT in whom at least one of the donor ZFPM1 gene and the recipient ACR gene is not mutated.

[0072] The method for diagnosing whether acute graft-versus-host disease occurs in patients after alloHCT based on whether the ZFPM1-ACR gene pair in the donor and the recipient is mutated is as follows: if the donor ZFPM1 gene is mutated and the recipient ACR gene is mutated, the patient does not develop or is a candidate for not developing acute graft-versus-host disease after alloHCT; if at least one of the donor ZFPM1 gene and the recipient ACR gene is not mutated, the patient develops or is a candidate for developing acute graft-versus-host disease after alloHCT.

[0073] The method for predicting the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation based on whether the ACTN2-OBSCN gene pair in the donor and the recipient is mutated is as follows: the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation with a mutation in the donor ACTN2 gene and a mutation in the recipient OBSCN gene is higher than that in patients after hematopoietic cell transplantation in which at least one of the donor ACTN2 gene and the recipient OBSCN gene is not mutated.

[0074] The method for diagnosing whether a patient develops acute graft-versus-host disease after hematopoietic cell transplantation based on whether the ACTN2-OBSCN gene pair in the donor and the recipient is mutated is as follows: if the donor's ACTN2 gene is mutated and the recipient's OBSCN gene is mutated, the patient develops or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation; if at least one of the donor's ACTN2 gene and the recipient's OBSCN gene is not mutated, the patient does not develop or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation.

[0075] The method for predicting the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation based on whether the CDH15-FRMD5 gene pair is mutated in the donor and the recipient is as follows: the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation in which both the donor CDH15 gene and the recipient FRMD5 gene are mutated is higher than that in patients after hematopoietic cell transplantation in which at least one of the donor CDH15 gene and the recipient FRMD5 gene is not mutated.

[0076] The method for diagnosing whether a patient develops acute graft-versus-host disease after hematopoietic cell transplantation based on whether the CDH15-FRMD5 gene pair in the donor and the recipient is mutated is as follows: if the donor CDH15 gene is mutated and the recipient FRMD5 gene is mutated, the patient develops or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation; if at least one of the donor CDH15 gene and the recipient FRMD5 gene is not mutated, the patient does not develop or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation.

[0077] The method for predicting the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation based on whether the CDH15-GPR161 gene pair is mutated in the donor and the recipient is as follows: the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation in which both the donor CDH15 gene is mutated and the recipient GPR161 gene is mutated is higher than that in patients after hematopoietic cell transplantation in which at least one of the donor CDH15 gene and the recipient GPR161 gene is not mutated.

[0078] The method for diagnosing whether a patient develops acute graft-versus-host disease after hematopoietic cell transplantation based on whether the CDH15-GPR161 gene pair in the donor and the recipient is mutated is as follows: if the donor CDH15 gene is mutated and the recipient GPR161 gene is mutated, the patient develops or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation; if at least one of the donor CDH15 gene and the recipient GPR161 gene is not mutated, the patient does not develop or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation.

[0079] The method for predicting the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation based on whether the CDH15-OBSCN gene pair is mutated in the donor and the recipient is as follows: the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation in which both the donor CDH15 gene is mutated and the recipient OBSCN gene is mutated is higher than that in patients after hematopoietic cell transplantation in which at least one of the donor CDH15 gene and the recipient OBSCN gene is not mutated.

[0080] The method for diagnosing whether a patient develops acute graft-versus-host disease after hematopoietic cell transplantation based on whether the CDH15-OBSCN gene pair in the donor and the recipient is mutated is as follows: if the donor CDH15 gene is mutated and the recipient OBSCN gene is mutated, the patient develops or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation; if at least one of the donor CDH15 gene and the recipient OBSCN gene is not mutated, the patient does not develop or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation.

[0081] The method for predicting the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation based on whether the GNRH2-FRMD5 gene pair is mutated in the donor and the recipient is as follows: the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation with a mutation in the donor GNRH2 gene and a mutation in the recipient FRMD5 gene is higher than that in patients after hematopoietic cell transplantation in which at least one of the donor GNRH2 gene and the recipient FRMD5 gene is not mutated.

[0082] The method for diagnosing whether a patient develops acute graft-versus-host disease after hematopoietic cell transplantation based on whether the GNRH2-FRMD5 gene pair in the donor and the recipient is mutated is as follows: if the donor GNRH2 gene is mutated and the recipient FRMD5 gene is mutated, the patient develops or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation; if at least one of the donor GNRH2 gene and the recipient FRMD5 gene is not mutated, the patient does not develop or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation.

[0083] The method for predicting the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation based on whether the GNRH2-KRT24 gene pair is mutated in the donor and the recipient is as follows: the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation with a mutation in the donor GNRH2 gene and a mutation in the recipient KRT24 gene is higher than that in patients after hematopoietic cell transplantation in which at least one of the donor GNRH2 gene and the recipient KRT24 gene is not mutated.

[0084] The method for diagnosing whether a patient develops acute graft-versus-host disease after hematopoietic cell transplantation based on whether the GNRH2-KRT24 gene pair in the donor and the recipient is mutated is as follows: if the donor GNRH2 gene is mutated and the recipient KRT24 gene is mutated, the patient develops or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation; if at least one of the donor GNRH2 gene and the recipient KRT24 gene is not mutated, the patient does not develop or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation.

[0085] The method for predicting the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation based on whether the GNRH2-MAGEE2 gene pair in the donor and the recipient is mutated is as follows: the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation with both the donor GNRH2 gene mutation and the recipient MAGEE2 gene mutation is higher than that in patients after hematopoietic cell transplantation in which at least one of the donor GNRH2 gene and the recipient MAGEE2 gene is not mutated.

[0086] The method for diagnosing whether a patient develops acute graft-versus-host disease after hematopoietic cell transplantation based on whether the GNRH2-MAGEE2 gene pair in the donor and the recipient is mutated is as follows: if the donor GNRH2 gene is mutated and the recipient MAGEE2 gene is mutated, the patient develops or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation; if at least one of the donor GNRH2 gene and the recipient MAGEE2 gene is not mutated, the patient does not develop or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation.

[0087] The method for predicting the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation based on whether the GNRH2-OBSCN gene pair in the donor and recipient is mutated is as follows: the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation with a mutation in the donor GNRH2 gene and a mutation in the recipient OBSCN gene is higher than that in patients after hematopoietic cell transplantation in which at least one of the donor GNRH2 gene and the recipient OBSCN gene is not mutated.

[0088] The method for diagnosing whether a patient develops acute graft-versus-host disease after hematopoietic cell transplantation based on whether the GNRH2-OBSCN gene pair in the donor and the recipient is mutated is as follows: if the donor GNRH2 gene is mutated and the recipient OBSCN gene is mutated, the patient develops or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation; if at least one of the donor GNRH2 gene and the recipient OBSCN gene is not mutated, the patient does not develop or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation.

[0089] The method for predicting the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation based on whether the GPSM1-OBSCN gene pair is mutated in the donor and the recipient is as follows: the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation in which both the donor GPSM1 gene and the recipient OBSCN gene are mutated is higher than that in patients after hematopoietic cell transplantation in which at least one of the donor GPSM1 gene and the recipient OBSCN gene is not mutated.

[0090] The method for diagnosing whether a patient develops acute graft-versus-host disease after hematopoietic cell transplantation based on whether the GPSM1-OBSCN gene pair in the donor and the recipient is mutated is as follows: if the donor GPSM1 gene is mutated and the recipient OBSCN gene is mutated, the patient develops or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation; if at least one of the donor GPSM1 gene and the recipient OBSCN gene is not mutated, the patient does not develop or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation.

[0091] The method for predicting the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation based on whether the HAPLN3-OBSCN gene pair is mutated in the donor and the recipient is as follows: the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation in which both the donor HAPLN3 gene and the recipient OBSCN gene are mutated is higher than that in patients after hematopoietic cell transplantation in which at least one of the donor HAPLN3 gene and the recipient OBSCN gene is not mutated.

[0092] The method for diagnosing whether a patient develops acute graft-versus-host disease after hematopoietic cell transplantation based on whether the HAPLN3-OBSCN gene pair in the donor and the recipient is mutated is as follows: if the donor HAPLN3 gene is mutated and the recipient OBSCN gene is mutated, the patient develops or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation; if at least one of the donor HAPLN3 gene and the recipient OBSCN gene is not mutated, the patient does not develop or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation.

[0093] The method for predicting the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation based on whether the LCN10-CD33 gene pair in the donor and the recipient is mutated is as follows: the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation with a mutation in the donor LCN10 gene and a mutation in the recipient CD33 gene is higher than that in patients after hematopoietic cell transplantation in which at least one of the donor LCN10 gene and the recipient CD33 gene is not mutated.

[0094] The method for diagnosing whether a patient develops acute graft-versus-host disease after hematopoietic cell transplantation based on whether the LCN10-CD33 gene pair in the donor and the recipient is mutated is as follows: if the donor LCN10 gene is mutated and the recipient CD33 gene is mutated, the patient develops or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation; if at least one of the donor LCN10 gene and the recipient CD33 gene is not mutated, the patient does not develop or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation.

[0095] The method for predicting the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation based on whether the MS4A12-ACTN2 gene pair in the donor and the recipient is mutated is as follows: the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation with a mutation in the donor MS4A12 gene and a mutation in the recipient ACTN2 gene is higher than that in patients after hematopoietic cell transplantation in which at least one of the donor MS4A12 gene and the recipient ACTN2 gene is not mutated.

[0096] The method for diagnosing whether a patient develops acute graft-versus-host disease after hematopoietic cell transplantation based on whether the MS4A12-ACTN2 gene pair in the donor and the recipient is mutated is as follows: if the donor MS4A12 gene is mutated and the recipient ACTN2 gene is mutated, the patient develops or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation; if at least one of the donor MS4A12 gene and the recipient ACTN2 gene is not mutated, the patient does not develop or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation.

[0097] The method for predicting the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation based on whether the MS4A12-ZNF211 gene pair is mutated in the donor and the recipient is as follows: the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation with mutations in the donor MS4A12 gene and mutations in the recipient ZNF211 gene is higher than that in patients after hematopoietic cell transplantation in which at least one of the donor MS4A12 gene and the recipient ZNF211 gene is not mutated.

[0098] The method for diagnosing whether a patient develops acute graft-versus-host disease after hematopoietic cell transplantation based on whether the MS4A12-ZNF211 gene pair in the donor and the recipient is mutated is as follows: if the donor MS4A12 gene is mutated and the recipient ZNF211 gene is mutated, the patient develops or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation; if at least one of the donor MS4A12 gene and the recipient ZNF211 gene is not mutated, the patient does not develop or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation.

[0099] The method for predicting the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation based on whether the RBMX-CD33 gene pair is mutated in the donor and the recipient is as follows: the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation in which both the donor RBMX gene is mutated and the recipient CD33 gene is mutated is higher than that in patients after hematopoietic cell transplantation in which at least one of the donor RBMX gene and the recipient CD33 gene is not mutated.

[0100] The method for diagnosing whether a patient develops acute graft-versus-host disease after hematopoietic cell transplantation based on whether the RBMX-CD33 gene pair in the donor and the recipient is mutated is as follows: if the donor RBMX gene is mutated and the recipient CD33 gene is mutated, the patient develops or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation; if at least one of the donor RBMX gene and the recipient CD33 gene is not mutated, the patient does not develop or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation.

[0101] The method for predicting the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation based on whether the SIRPB1-ACTN2 gene pair in the donor and the recipient is mutated is as follows: the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation with a mutation in the donor SIRPB1 gene and a mutation in the recipient ACTN2 gene is higher than that in patients after hematopoietic cell transplantation in which at least one of the donor SIRPB1 gene and the recipient ACTN2 gene is not mutated.

[0102] The method for diagnosing whether a patient develops acute graft-versus-host disease after hematopoietic cell transplantation based on whether the SIRPB1-ACTN2 gene pair in the donor and the recipient is mutated is as follows: if the donor's SIRPB1 gene is mutated and the recipient's ACTN2 gene is mutated, the patient develops or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation; if at least one of the donor's SIRPB1 gene and the recipient's ACTN2 gene is not mutated, the patient does not develop or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation.

[0103] The method for predicting the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation based on whether the SIRPB1-OBSCN gene pair in the donor and recipient is mutated is as follows: the risk of acute graft-versus-host disease in patients after hematopoietic cell transplantation with a mutation in the donor SIRPB1 gene and a mutation in the recipient OBSCN gene is higher than that in patients after hematopoietic cell transplantation in which at least one of the donor SIRPB1 gene and the recipient OBSCN gene is not mutated.

[0104] The method for diagnosing whether a patient develops acute graft-versus-host disease after hematopoietic cell transplantation based on whether the SIRPB1-OBSCN gene pair in the donor and the recipient is mutated is as follows: if the donor's SIRPB1 gene is mutated and the recipient's OBSCN gene is mutated, the patient develops or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation; if at least one of the donor's SIRPB1 gene and the recipient's OBSCN gene is not mutated, the patient does not develop or is a candidate to develop acute graft-versus-host disease after hematopoietic cell transplantation.

[0105] In practical applications, any two or three or more gene pairs (including 35 gene pairs) among the above 35 gene pairs can also be used to predict the risk of acute graft-versus-host disease in patients after alloHCT of different modes or to diagnose whether acute graft-versus-host disease occurs in patients after alloHCT. When multiple gene pairs are used for joint prediction or diagnosis, each gene pair needs to meet its own judgment criteria for prediction or diagnosis, otherwise the prediction or diagnosis method is invalid.

[0106] When the 35 gene pairs are used for prediction or diagnosis, in addition to referring to the judgment criteria of each gene pair mentioned above for prediction or diagnosis, prediction or diagnosis can also be performed by comparing the cumulative score of the mutant gene pairs with the threshold.

[0107] Specifically,

[0108] The method for predicting the risk of acute graft-versus-host disease in patients after alloHCT based on the cumulative score and threshold of mutation gene pairs is as follows: the risk of acute graft-versus-host disease in patients after alloHCT whose cumulative score of gene mutation pairs is greater than or equal to the threshold is higher than that in patients after alloHCT whose cumulative score of gene mutation pairs is less than the threshold.

[0109] Whether acute graft-versus-host disease occurs in patients after alloHCT is diagnosed based on the cumulative score and threshold of the mutation gene pair: if the cumulative score of the gene mutation is greater than or equal to the threshold, the patient has developed or is a candidate for developing acute graft-versus-host disease after alloHCT; if the cumulative score of the gene mutation is less than the threshold, the patient has not developed or is a candidate for developing acute graft-versus-host disease after alloHCT.

[0110] The cumulative score of the mutant gene pair is obtained according to the following method: detecting the mutation status of the protective mutant gene pair in the peripheral blood of the patient and the donor, and obtaining the cumulative score of the protective mutant gene pair; detecting the mutation status of the promoting mutant gene pair in the peripheral blood of the patient and the donor, and obtaining the cumulative score of the promoting mutant gene pair; the sum of the cumulative score of the protective mutant gene pair and the cumulative score of the promoting mutant gene pair is the cumulative score of the mutant gene pair of the patient;

[0111] The protective mutant gene pairs are ADAMTS7-APOL4, AGAP3-ACR, ANAPC1-ACR, CARD8-SIRPB1, DCAF15-ACR, DENND3-ACR, FRMD5-MYCT1, HAPLN3-ACR, HRNR-ACR, MAGEE2-ACR, MYCT1-KRTAP10-1, MYCT1-OAS1, NUDT1-ACR, OAS1-ACR, PABPC3-ACR, SIRPA-ACR, SLC35G6-ACR, TLR8-ACR, and ZFPM1-ACR;

[0112] The promoting mutation gene pairs are ACTN2-OBSCN, CDH15-FRMD5, CDH15-GPR161, CDH15-OBSCN, GNRH2-FRMD5, GNRH2-KRT24, GNRH2-MAGEE2, GNRH2-OBSCN, GPSM1-OBSCN, HAPLN3-OBSCN, LCN10-CD33, MS4A12-ACTN2, MS4A12-ZNF211, RBMX-CD33, SIRPB1-ACTN2, SIRPB1-OBSCN;

[0113] The cumulative score of the protective mutation gene pairs is calculated as follows: if none of the 19 protective mutation gene pairs in the peripheral blood of the patient and the donor is mutated, it is recorded as 0; if one protective mutation gene pair (any one of the 19 protective mutation gene pairs) is mutated in the peripheral blood of the patient and the donor, it is recorded as -1; if two protective mutation gene pairs (any two of the 19 protective mutation gene pairs) are mutated in the peripheral blood of the patient and the donor, it is recorded as -2; and so on, if 19 protective mutation gene pairs are mutated in the peripheral blood of the patient and the donor, it is recorded as -19;

[0114] The calculation method for the cumulative score of the promoting mutation gene pairs is as follows: if none of the 16 promoting mutation gene pairs in the peripheral blood of the patient and its donor are mutated, it is recorded as 0; if one promoting mutation gene pair (any one of the 16 promoting mutation gene pairs) in the peripheral blood of the patient and its donor is mutated, it is recorded as 1; if two promoting mutation gene pairs (any two of the 16 promoting mutation gene pairs) in the peripheral blood of the patient and its donor are mutated, it is recorded as 2; and so on, if 16 promoting mutation gene pairs in the peripheral blood of the patient and its donor are mutated, it is recorded as 16.

[0115] In practical applications, the risk of acute graft-versus-host disease after alloHCT of different modes can be predicted according to the above method, and then the pre-alloHCT donors can be screened according to the risk of acute graft-versus-host disease, or other clinical indicators can be combined to assist in the screening of pre-alloHCT donors, so as to optimize the selection of pre-alloHCT donors and improve the survival rate and quality of life of patients before alloHCT.

[0116] Mutation in any of the above gene pairs means that both the gene corresponding to the donor (i.e. the first gene before the “-” in the above text) and the gene corresponding to the recipient (i.e. the second gene after the “-” in the above text) in the gene pair are mutated.

[0117] Mutations in any of the above genes include any base mutations that can change the amino acid sequence of the protein encoded by the gene. The base mutations include base insertions, base deletions, base substitutions, etc. The number of base mutations includes 1 or 2 or more base mutations.

[0118] The fact that any of the above genes have not undergone mutations includes the fact that there are no base mutations (completely identical to the wild-type sequence of the gene) or any base mutations that do not change the amino acid sequence of the protein encoded by the gene.

[0119] Any of the above thresholds may specifically be 1.5 or 3.5.

[0120] In any of the above applications or kits, the donor-recipient refers to a donor and a recipient. The donor is a healthy individual who provides hematopoietic cells before alloHCT, and the recipient is a patient before alloHCT.

[0121] In any of the above applications or kits, the alloHCT is allogeneic hematopoietic cell transplantation.

[0122] The allogeneic hematopoietic cell transplantation includes HLA haplotype-matched allogeneic hematopoietic cell transplantation and HLA fully-matched allogeneic hematopoietic cell transplantation.

[0123] The patient before alloHCT may be a patient with acute myeloid leukemia, a patient with acute lymphoid leukemia, a patient with chronic myeloid leukemia, a patient with chronic lymphoid leukemia, a patient with multiple myeloma, a patient with myelodysplastic syndrome, or a patient with primary systemic amyloidosis.

[0124] The use of ADAMTS7, APOL4, AGAP3, ACR, ANAPC1, CARD8, SIRPB1, DCAF15, DENND3, FRMD5, MYCT1, HAPLN3, HRNR, MAGEE2, KRTAP10-1, OAS1, NUDT1, PABPC3, SIRPA, SLC35G6, TLR8, ZFPM1, ACTN2, OBSCN, CDH15, GPR161, GNRH2, KRT24, GPSM1, LCN10, CD33, MS4A12, ZNF211 and / or RBMX as targets in the preparation of products for preventing and / or treating acute graft-versus-host disease also falls within the scope of protection of the present invention.

[0125] The IDs of any of the above-mentioned ADAMTS7, APOL4, AGAP3, ACR, ANAPC1, CARD8, SIRPB1, DCAF15, DENND3, FRMD5, MYCT1, HAPLN3, HRNR, MAGEE2, KRTAP10-1, OAS1, NUDT1, PABPC3, SIRPA, SLC35G6, TLR8, ZFPM1, ACTN2, OBSCN, CDH15, GPR161, GNRH2, KRT24, GPSM1, LCN10, CD33, MS4A12, ZNF211, and RBMX in the NCBI gene library are shown in Table 1 below.

[0126] The present invention has discovered gene combinations with predictive and diagnostic value for the occurrence of aGVHD after alloHCT in different modes (HLA haplotype matching and HLA full matching) through whole genome sequencing and target gene capture sequencing. On the one hand, by detecting the gene combination mutations of donors and recipients for comprehensive judgment and evaluation, the selection of donors before alloHCT is optimized, and the survival rate and quality of life of patients are improved; on the other hand, the risk of aGVHD after alloHCT in recipients can be predicted by detecting the gene combination mutations of donors and recipients, providing a basis for potential diagnosis of aGVHD; thirdly, the present invention provides a new genetic basis for revealing the pathogenesis of aGVHD and provides a new drug target for the treatment of aGVHD. BRIEF DESCRIPTION OF THE DRAWINGS

[0127] Figure 1 The LOF mutation gene pairs of 212 patients and their donors were scored cumulatively.

[0128] Figure 2 The ROC curve of the LOF mutation gene pairs of 212 patients and their donors was used to diagnose aGVHD.

[0129] Figure 3The LOF mutation gene pairs of 353 patients and their donors were cumulatively scored.

[0130] Figure 4 The ROC curve of the LOF mutation gene pairs of 353 patients and their donors was used to diagnose aGVHD.

[0131] Figure 5 The LOF mutation gene pairs of 565 patients and their donors were cumulatively scored.

[0132] Figure 6 The ROC curve of the LOF mutation gene pairs for 565 patients and their donors was used to diagnose aGVHD. DETAILED DESCRIPTION

[0133] The present invention is further described in detail below in conjunction with specific embodiments, and the examples provided are only for illustrating the present invention, rather than for limiting the scope of the present invention. The examples provided below can be used as a guide for further improvements by those of ordinary skill in the art, and do not constitute a limitation of the present invention in any way.

[0134] The experimental methods in the following examples, unless otherwise specified, are all conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials, reagents, etc. used in the following examples, unless otherwise specified, can all be obtained from commercial channels.

[0135] The HLA haplotype matching involved in the present invention means that the HLA-A, HLA-B, and HLA-DRB1 alleles between the related donor and recipient are the same on the same chromosome, but at least one of the above alleles on another chromosome is different (i.e. 3 / 6, 4 / 6, or 5 / 6 matching).

[0136] The HLA full match involved in the present invention means that the HLA-A, HLA-B, and HLA-DRB1 alleles between the sibling donor and recipient are the same on both chromosomes (i.e., 6 / 6 match).

[0137] Example 1. Acquisition and verification of LoF mutation gene pairs associated with aGVHD

[0138] 1. Based on the HLA haplotype-matched alloHCT clinical cohort, LoF mutation gene pairs associated with aGVHD were found

[0139] 1. Research subjects

[0140] The research subjects of this example are as follows: peripheral blood samples of 106 patients with aGVHD before hematopoietic cell transplantation and their donor peripheral blood samples (defined as the aGVHD positive group), and peripheral blood samples of 106 patients without aGVHD before hematopoietic cell transplantation and their donor peripheral blood samples (defined as the aGVHD negative group).

[0141] Inclusion criteria: 1. Patients underwent HLA haplotype-matched alloHCT at Peking University People's Hospital in 2018 and 2019; 2. Patients were diagnosed with hematological malignancies before hematopoietic cell transplantation, including acute myeloid leukemia, acute lymphoid leukemia, chronic myeloid leukemia, chronic lymphoid leukemia, multiple myeloma, myelodysplastic syndrome, and primary systemic amyloidosis; 3. The ages of patients and donors were between 14 and 65 years old; 4. The aGVHD-positive group included patients diagnosed with aGVHD 2-4 within 100 days after hematopoietic cell transplantation; the aGVHD-negative group included patients who did not develop aGVHD symptoms within 100 days after hematopoietic cell transplantation, and were paired 1:1 with the aGVHD-positive group. There was no statistical difference in age and gender between the two groups.

[0142] Exclusion criteria: 1. Patients diagnosed with non-acute promyelocytic leukemia or aplastic anemia before hematopoietic cell transplantation; 2. Patients who were not undergoing alloHCT for the first time; 3. Patients who died within three months after hematopoietic cell transplantation and the cause of death was not diagnosed as aGVHD.

[0143] 2. Sample preparation

[0144] The genomic DNA was extracted from the collected peripheral blood samples using the phenol-chloroform method to test the purity, concentration, integrity and contamination of the DNA. The qualified samples were sequenced for genome sequencing, and the experimental process was carried out according to the library preparation steps and sequencing steps provided by Illumina.

[0145] 3. Whole genome sequencing

[0146] The library was constructed using the human whole exome sequence capture kit Truseq Nano, and hybridized with probes for capture and enrichment. The samples were then sequenced using the Illumina Novaseq S4 PE150 high-throughput sequencing platform, with an average sequencing depth of 35x for each sample.

[0147] 4. Analysis of sequencing results

[0148] FastQC software was used to evaluate the quality of the output raw data, and BWA (parameter-M) software was used to align the measured data with the reference genome. All analyses were performed based on the human reference genome hg38. The mutations that affect protein structure annotated by ANNOVAR were defined as loss-of-function mutations (LOF mutations) in gene expression proteins in this study. The ANNOVAR annotations included "stopgain", "startloss", "frameshift insertion", "frameshift deletion" and "frameshift block substitution". When LoF mutations occur in two genes in a pair of donors and recipients (such as mutations in gene A in the donor and mutations in gene B in the recipient), the donor A gene and the recipient B gene are defined as a LoF mutation gene pair.

[0149] A total of 35 LoF mutation gene pairs associated with aGVHD were obtained by screening in the HLA haplotype alloHCT clinical cohort, as shown in Table 1.

[0150] 5. Correlation analysis between LoF mutation gene pairs and aGVHD

[0151] In the HLA haplotype-matched alloHCT cohort, each LoF mutation gene pair was analyzed for correlation with the occurrence of aGVHD.

[0152] The results are shown in Table 1. It can be seen from Table 1 that there are significant statistical differences between the aGVHD-positive group and the aGVHD-negative group in the 35 LOF mutation gene pairs, indicating that the 35 LOF mutation gene pairs are significantly correlated with the occurrence of aGVHD after HLA haplotype-matched alloHCT. Among them, 19 LOF mutation gene pairs are significantly negatively correlated with the occurrence of aGVHD (marked as aGVHD- in the table), and 16 LOF mutation gene pairs are significantly positively correlated with the occurrence of aGVHD (marked as aGVHD+ in the table).

[0153] Table 1

[0154]

[0155] 2. Validation of LoF mutation gene pairs associated with aGVHD in the HLA-matched alloHCT clinical cohort

[0156] 1. Research subjects

[0157] The research subjects of this example are as follows: peripheral blood samples of 71 patients with aGVHD before hematopoietic cell transplantation and their donor peripheral blood samples (defined as the aGVHD positive group), and peripheral blood samples of 282 patients without aGVHD before hematopoietic cell transplantation and their donor peripheral blood samples (defined as the aGVHD negative group).

[0158] Inclusion criteria: 1. Patients underwent HLA-matched alloHCT at Peking University People's Hospital from 2018 to 2021; 2. Patients were diagnosed with hematological malignancies before hematopoietic cell transplantation, including acute myeloid leukemia, acute lymphoid leukemia, chronic myeloid leukemia, chronic lymphoid leukemia, multiple myeloma, myelodysplastic syndrome, and primary systemic amyloidosis; 3. Both patients and donors were between 14 and 65 years old.

[0159] Exclusion criteria: 1. Patients diagnosed with non-acute promyelocytic leukemia or aplastic anemia before hematopoietic cell transplantation; 2. Patients who were not undergoing alloHCT for the first time; 3. Patients who died within three months after hematopoietic cell transplantation and the cause of death was not diagnosed as aGVHD.

[0160] 2. Sample preparation

[0161] The peripheral blood samples collected were used to extract genomic DNA and test the purity, concentration, integrity and contamination of the DNA. The qualified samples were then tested for the next step.

[0162] 3. Probe sequencing

[0163] 200 ng of genomic DNA was randomly fragmented into 150-200 bp average length using enzyme digestion, and then end-repaired and flattened with an A base added to the 3' end to form a sticky end. Then it was connected to the adapter containing the target gene-specific barcode sequence. Magnetic bead screening was used to remove incomplete connection products and adapter self-connection products. PCR amplification was performed using universal primers complementary to the adapter sequence to form a sequencing library. Then, the library was used to generate the sequencing library. The pool probes were hybridized in the hybridization buffer, and the hybridized target fragments were captured using magnetic beads and separated and purified. The captured DNA fragments were amplified by PCR and the PCR products were purified. The quality control of the captured library was then performed, including Qubit concentration determination and 2100 Bioanalyzer fragment length determination. Finally, PE150 high-throughput sequencing was performed on the NovaSeq6000 sequencer according to Illumina's standard process.

[0164] 4. Analysis of test results

[0165] FastQC software was used to evaluate the quality of the raw data, and BWA (parameter-M) software was used for genome alignment. All analyses were performed based on the human reference genome hg38.

[0166] 5. Correlation analysis between LoF mutation gene pairs and aGVHD

[0167] In the HLA-matched alloHCT cohort, each LoF mutation gene pair was analyzed for correlation with the occurrence of aGVHD.

[0168] The results are shown in Table 1. It can be seen from Table 1 that the 35 LOF mutation gene pairs screened in step 1 showed significant statistical differences between the aGVHD positive group and the aGVHD negative group, indicating that the 35 LOF mutation gene pairs were still significantly correlated with the occurrence of aGVHD after HLA fully matched alloHCT. Among them, 19 LOF mutation gene pairs were significantly negatively correlated with the occurrence of aGVHD (labeled as aGVHD- in Table 1), and 16 LOF mutation gene pairs were significantly positively correlated with the occurrence of aGVHD (labeled as aGVHD+ in Table 1). This result is consistent with the analysis results in the HLA haplotype-matched alloHCT cohort.

[0169] 3. Specific mutation status of 35 LOF mutation gene pairs in HLA haplotype-matched and HLA-matched alloHCT cohorts

[0170] 1. ADAMTS7-APOL4 gene mutation

[0171] Among the 212 donor-recipient pairs in the HLA haplotype-matched cohort, 8 cases had ADAMTS7-APOL4 gene mutations, including 1 case in the aGVHD-positive group and 7 cases in the aGVHD-negative group. The aGVHD grouping and gene pair mutations are shown in Table 2. After chi-square test analysis, P=0.0306, indicating a statistically significant difference between the two groups.

[0172] Table 2

[0173]

[0174] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0175] Among the 353 donor-recipient pairs in the HLA fully matched cohort, there were 119 cases, including 16 cases in the aGVHD-positive group and 103 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 3. After chi-square test analysis, P=0.0258, there was a statistically significant difference between the two groups.

[0176] Table 3

[0177]

[0178] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0179] 2. AGAP3-ACR gene mutation

[0180] Among the 212 donor-recipient pairs in the HLA haplotype-matched cohort, 6 cases had AGAP3-ACR gene mutations, including 0 cases in the aGVHD-positive group and 6 cases in the aGVHD-negative group. The aGVHD grouping and gene pair mutations are shown in Table 4. After chi-square test analysis, P=0.0130, there was a statistically significant difference between the two groups.

[0181] Table 4

[0182]

[0183] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0184] Among the 353 donor-recipient pairs in the HLA fully matched cohort, 26 cases had AGAP3-ACR gene mutations, including 1 case in the aGVHD-positive group and 25 cases in the aGVHD-negative group. The aGVHD grouping and gene pair mutations are shown in Table 5. After chi-square test analysis, P=0.0316, indicating a statistically significant difference between the two groups.

[0185] Table 5

[0186]

[0187] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0188] ANAPC1-ACR gene mutation

[0189] Among the 212 donor-recipient pairs in the HLA haplotype-matched cohort, 7 cases had ANAPC1-ACR gene mutations, including 0 cases in the aGVHD-positive group and 7 cases in the aGVHD-negative group. The aGVHD grouping and gene pair mutations are shown in Table 6. After chi-square test analysis, P=0.0071, indicating a statistically significant difference between the two groups.

[0190] Table 6

[0191]

[0192] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0193] Among the 353 donor-recipient pairs in the HLA fully matched cohort, 23 cases had ANAPC1-ACR gene mutations, including 1 case in the aGVHD-positive group and 22 cases in the aGVHD-negative group. The aGVHD grouping and gene pair mutations are shown in Table 7. After chi-square test analysis, P=0.0511, indicating a statistically significant difference between the two groups.

[0194] Table 7

[0195]

[0196]

[0197] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0198] 4. CARD8-SIRPB1 gene mutation

[0199] Among the 212 donor-recipient pairs in the HLA haplotype-matched cohort, 50 cases had CARD8-SIRPB1 gene mutations, including 18 cases in the aGVHD-positive group and 32 cases in the aGVHD-negative group. The aGVHD grouping and gene pair mutations are shown in Table 8. After chi-square test analysis, P=0.0235, indicating a statistically significant difference between the two groups.

[0200] Table 8

[0201]

[0202] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0203] Among the 353 donor-recipient pairs in the HLA fully matched cohort, there were 242 cases of CARD8-SIRPB1 gene mutations, including 40 cases in the aGVHD-positive group and 202 cases in the aGVHD-negative group. The aGVHD grouping and gene pair mutations are shown in Table 9. After chi-square test analysis, P=0.0131, there was a statistically significant difference between the two groups.

[0204] Table 9

[0205]

[0206] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0207] 5. DCAF15-ACR gene mutation

[0208] Among the 212 donor-recipient pairs in the HLA haplotype-matched cohort, 11 cases had DCAF15-ACR gene mutations, including 2 cases in the aGVHD-positive group and 9 cases in the aGVHD-negative group. The aGVHD grouping and gene pair mutations are shown in Table 10. After chi-square test analysis, P=0.0302, indicating a statistically significant difference between the two groups.

[0209] Table 10

[0210]

[0211]

[0212] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0213] Among the 353 donor-recipient pairs in the HLA fully matched cohort, 20 cases had DCAF15-ACR gene mutations, including 0 cases in the aGVHD-positive group and 20 cases in the aGVHD-negative group. The aGVHD grouping and gene pair mutations are shown in Table 11. After chi-square test analysis, P=0.0209, indicating a statistically significant difference between the two groups.

[0214] Table 11

[0215]

[0216] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0217] 6. DENND3-ACR gene mutation

[0218] Among the 212 donor-recipient pairs in the HLA haplotype-matched cohort, 12 cases had DENND3-ACR gene mutations, including 2 cases in the aGVHD-positive group and 10 cases in the aGVHD-negative group. The aGVHD grouping and gene pair mutations are shown in Table 12. After chi-square test analysis, P=0.0174, indicating a statistically significant difference between the two groups.

[0219] Table 12

[0220]

[0221] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0222] Among the 353 donor-recipient pairs in the HLA fully matched cohort, 27 cases had DENND3-ACR gene mutations, including 1 case in the aGVHD-positive group and 26 cases in the aGVHD-negative group. The aGVHD grouping and gene pair mutations are shown in Table 13. After chi-square test analysis, P=0.0269, indicating a statistically significant difference between the two groups.

[0223] Table 13

[0224]

[0225]

[0226] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0227] 7. FRMD5-MYCT1 gene mutation

[0228] Among the 212 donor-recipient pairs in the HLA haplotype-matched cohort, 49 cases had FRMD5-MYCT1 gene mutations, including 16 cases in the aGVHD-positive group and 33 cases in the aGVHD-negative group. The aGVHD grouping and gene pair mutation status are shown in Table 14. After chi-square test analysis, P=0.0056, indicating a statistically significant difference between the two groups.

[0229] Table 14

[0230]

[0231] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0232] Among the 353 donor-recipient pairs in the HLA fully matched cohort, 67 cases had FRMD5-MYCT1 gene mutations, including 7 cases in the aGVHD-positive group and 60 cases in the aGVHD-negative group. The aGVHD grouping and gene pair mutation status are shown in Table 15. After chi-square test analysis, P=0.0283, there was a statistically significant difference between the two groups.

[0233] Table 15

[0234]

[0235] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0236] 8. HAPLN3-ACR gene mutation

[0237] Among the 212 donor-recipient pairs in the HLA haplotype-matched cohort, 8 cases had HAPLN3-ACR gene mutations, including 1 case in the aGVHD-positive group and 7 cases in the aGVHD-negative group. The aGVHD grouping and gene pair mutation status are shown in Table 16. After chi-square test analysis, P=0.0306, indicating a statistically significant difference between the two groups.

[0238] Table 16

[0239]

[0240] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0241] Among the 212 donor-recipient pairs in the HLA fully matched cohort, 16 cases had HAPLN3-ACR gene mutations, including 0 cases in the aGVHD-positive group and 16 cases in the aGVHD-negative group. The aGVHD grouping and gene pair mutations are shown in Table 17. After chi-square test analysis, P=0.0400, indicating a statistically significant difference between the two groups.

[0242] Table 17

[0243]

[0244] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0245] 9. HRNR-ACR gene mutation

[0246] Among the 212 donor-recipient pairs in the HLA haplotype-matched cohort, 12 cases had HRNR-ACR gene mutations, including 2 cases in the aGVHD-positive group and 10 cases in the aGVHD-negative group. The aGVHD grouping and gene pair mutations are shown in Table 18. After chi-square test analysis, P=0.0174, indicating a statistically significant difference between the two groups.

[0247] Table 18

[0248]

[0249] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0250] Among the 353 donor-recipient pairs in the HLA fully matched cohort, 27 cases had HRNR-ACR gene mutations, including 1 case in the aGVHD-positive group and 26 cases in the aGVHD-negative group. The aGVHD grouping and gene pair mutations are shown in Table 19. After chi-square test analysis, P=0.0269, indicating a statistically significant difference between the two groups.

[0251] Table 19

[0252]

[0253] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0254] 10. MAGEE2-ACR gene mutation

[0255] Among the 212 donor-recipient pairs in the HLA haplotype-matched cohort, there were 11 cases of MAGEE2-ACR gene mutation, including 2 cases in the aGVHD-positive group and 9 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 20. After chi-square test analysis, P=0.0302, there was a statistically significant difference between the two groups.

[0256] Table 20

[0257]

[0258] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0259] Among the 353 donor-recipient pairs in the HLA fully matched cohort, 25 cases had MAGEE2-ACR gene mutations, including 1 case in the aGVHD-positive group and 24 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 21. After chi-square test analysis, P=0.0371, and there was a statistically significant difference between the two groups.

[0260] Table 21

[0261]

[0262] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0263] 11. MYCT1-KRTAP10-1 gene mutation

[0264] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, 61 cases had MAGEE2-ACR gene mutations, including 24 cases in the aGVHD-positive group and 37 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 22. After chi-square test analysis, P=0.0486, and there was a statistically significant difference between the two groups.

[0265] Table 22

[0266]

[0267] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0268] Among the 353 donor-recipient pairs in the HLA fully matched cohort, 83 cases had MAGEE2-ACR gene mutations, including 8 cases in the aGVHD-positive group and 75 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 23. After chi-square test analysis, P=0.0065, there was a statistically significant difference between the two groups.

[0269] Table 23

[0270]

[0271] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0272] 12. MYCT1-OAS1 gene mutation

[0273] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, 68 cases had MYCT1-OAS1 gene mutations, including 25 cases in the aGVHD-positive group and 43 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 24. After chi-square test analysis, P=0.0081, there was a statistically significant difference between the two groups.

[0274] Table 24

[0275]

[0276]

[0277] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0278] Among the 353 donor-recipient pairs in the HLA fully matched cohort, 99 cases had MYCT1-OAS1 gene mutations, including 13 cases in the aGVHD-positive group and 86 cases in the aGVHD-negative group. The aGVHD grouping and gene pair mutation status are shown in Table 25. After chi-square test analysis, P=0.0410, indicating a statistically significant difference between the two groups.

[0279] Table 25

[0280]

[0281] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0282] 13. NUDT1-ACR gene mutation

[0283] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, 12 cases had NUDT1-ACR gene mutations, including 2 cases in the aGVHD-positive group and 10 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 26. After chi-square test analysis, P=0.0174, there was a statistically significant difference between the two groups.

[0284] Table 26

[0285]

[0286] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0287] Among the 353 donor-recipient pairs in the HLA fully matched cohort, 27 cases had NUDT1-ACR gene mutations, including 1 case in the aGVHD-positive group and 26 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 27. After chi-square test analysis, P=0.0269, there was a statistically significant difference between the two groups.

[0288] Table 27

[0289]

[0290]

[0291] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0292] 14. OAS1-ACR gene mutation

[0293] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, 12 cases had OAS1-ACR gene mutations, including 2 cases in the aGVHD-positive group and 10 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 28. After chi-square test analysis, P=0.0174, there was a statistically significant difference between the two groups.

[0294] Table 28

[0295]

[0296] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0297] Among the 353 donor-recipient pairs in the HLA fully matched cohort, 25 cases had OAS1-ACR gene mutations, including 1 case in the aGVHD-positive group and 24 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 29. After chi-square test analysis, P=0.0371, and there was a statistically significant difference between the two groups.

[0298] Table 29

[0299]

[0300] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0301] 15. PABPC3-ACR gene mutation

[0302] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, 12 cases had PABPC3-ACR gene mutations, including 2 cases in the aGVHD-positive group and 10 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 30. After chi-square test analysis, P=0.0174, there was a statistically significant difference between the two groups.

[0303] Table 30

[0304]

[0305]

[0306] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0307] Among the 353 donor-recipient pairs in the HLA fully matched cohort, 26 cases had PABPC3-ACR gene mutations, including 1 case in the aGVHD-positive group and 25 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 31. After chi-square test analysis, P=0.0316, and there was a statistically significant difference between the two groups.

[0308] Table 31

[0309]

[0310] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0311] 16. SIRPA-ACR gene mutation

[0312] Among the 212 donor-recipient pairs in the HLA haplotype-matched cohort, there were 11 cases of SIRPA-ACR gene mutations, including 2 cases in the aGVHD-positive group and 9 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 32. After chi-square test analysis, P=0.0302, there was a statistically significant difference between the two groups.

[0313] Table 32

[0314]

[0315] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0316] Among the 353 donor-recipient pairs in the HLA fully matched cohort, there were 24 cases of SIRPA-ACR gene mutations, including 1 case in the aGVHD-positive group and 23 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 33. After chi-square test analysis, P=0.0435, there was a statistically significant difference between the two groups.

[0317] Table 33

[0318]

[0319] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0320] 17. SLC35G6-ACR gene mutation

[0321] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, 12 cases had SLC35G6-ACR gene mutations, including 2 cases in the aGVHD-positive group and 10 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 34. After chi-square test analysis, P=0.0174, there was a statistically significant difference between the two groups.

[0322] Table 34

[0323]

[0324] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0325] Among the 353 donor-recipient pairs in the HLA fully matched cohort, 27 cases had SLC35G6-ACR gene mutations, including 1 case in the aGVHD-positive group and 26 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 35. After chi-square test analysis, P=0.0269, there was a statistically significant difference between the two groups.

[0326] Table 35

[0327]

[0328] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0329] 18. TLR8-ACR gene mutation

[0330] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, there were 11 cases of TLR8-ACR gene mutation, including 2 cases in the aGVHD-positive group and 9 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 36. After chi-square test analysis, P=0.0302, there was a statistically significant difference between the two groups.

[0331] Table 36

[0332]

[0333] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0334] Among the 353 pairs of donors and recipients in the HLA fully matched cohort, there were 22 cases of TLR8-ACR gene mutation, including 0 cases in the aGVHD-positive group and 22 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 37. After chi-square test analysis, P=0.0151, there was a statistically significant difference between the two groups.

[0335] Table 37

[0336]

[0337] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0338] 19. ZFPM1-ACR gene mutation

[0339] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, 12 cases had ZFPM1-ACR gene mutations, including 2 cases in the aGVHD-positive group and 10 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 38. After chi-square test analysis, P=0.0174, there was a statistically significant difference between the two groups.

[0340] Table 38

[0341]

[0342] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0343] Among the 353 donor-recipient pairs in the HLA fully matched cohort, 27 cases had ZFPM1-ACR gene mutations, including 1 case in the aGVHD-positive group and 26 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 39. After chi-square test analysis, P=0.0269, and there was a statistically significant difference between the two groups.

[0344] Table 39

[0345]

[0346] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0347] 20. ACTN2-OBSCN gene mutation

[0348] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, there were 4 cases of ACTN2-OBSCN gene mutation, including 4 cases in the aGVHD-positive group and 0 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 40. After chi-square test analysis, P=0.0435, there was a statistically significant difference between the two groups.

[0349] Table 40

[0350]

[0351] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0352] Among the 353 donor-recipient pairs in the HLA fully matched cohort, there were 10 cases of ACTN2-OBSCN gene mutation, including 5 cases in the aGVHD positive group and 5 cases in the aGVHD negative group; the aGVHD grouping and gene pair mutation status are shown in Table 41. After chi-square test analysis, P=0.0168, there was a statistically significant difference between the two groups.

[0353] Table 41

[0354]

[0355] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0356] 21. CDH15-FRMD5 gene mutation

[0357] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, 38 cases had CDH15-FRMD5 gene mutations, including 25 cases in the aGVHD-positive group and 13 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 42. After chi-square test analysis, P=0.0317, and there was a statistically significant difference between the two groups.

[0358] Table 42

[0359]

[0360] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0361] Among the 353 donor-recipient pairs in the HLA fully matched cohort, 68 cases had CDH15-FRMD5 gene mutations, including 20 cases in the aGVHD-positive group and 48 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 43. After chi-square test analysis, P=0.0333, there was a statistically significant difference between the two groups.

[0362] Table 43

[0363]

[0364] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0365] 22. CDH15-GPR161 gene mutation

[0366] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, 5 cases had CDH15-GPR161 gene mutations, including 5 cases in the aGVHD-positive group and 0 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 44. After chi-square test analysis, P=0.0236, and there was a statistically significant difference between the two groups.

[0367] Table 44

[0368]

[0369]

[0370] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0371] Among the 353 donor-recipient pairs in the HLA fully matched cohort, there were 11 cases of CDH15-GPR161 gene mutations, including 6 cases in the aGVHD-positive group and 5 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 45. After chi-square test analysis, P=0.0038, there was a statistically significant difference between the two groups.

[0372] Table 45

[0373]

[0374] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0375] 23. CDH15-OBSCN gene mutation

[0376] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, 35 cases had CDH15-OBSCN gene mutations, including 27 cases in the aGVHD-positive group and 8 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 46. After chi-square test analysis, P=0.0004, there was a statistically significant difference between the two groups.

[0377] Table 46

[0378]

[0379] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0380] Among the 353 donor-recipient pairs in the HLA fully matched cohort, 70 cases had CDH15-OBSCN gene mutations, including 20 cases in the aGVHD-positive group and 50 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 47. After chi-square test analysis, P=0.0486, and there was a statistically significant difference between the two groups.

[0381] Table 47

[0382]

[0383]

[0384] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0385] 24. GNRH2-FRMD5 gene mutation

[0386] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, 71 cases had GNRH2-FRMD5 gene mutations, including 43 cases in the aGVHD-positive group and 28 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 48. After chi-square test analysis, P=0.0290, there was a statistically significant difference between the two groups.

[0387] Table 48

[0388]

[0389] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0390] Among the 353 donor-recipient pairs in the HLA fully matched cohort, there were 124 cases of GNRH2-FRMD5 gene mutations, including 33 cases in the aGVHD-positive group and 91 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 49. After chi-square test analysis, P=0.0250, there was a statistically significant difference between the two groups.

[0391] Table 49

[0392]

[0393] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0394] 25. GNRH2-KRT24 gene mutation

[0395] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, 115 cases had GNRH2-KRT24 gene mutations, including 67 cases in the aGVHD-positive group and 48 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 50. After chi-square test analysis, P=0.0088, and there was a statistically significant difference between the two groups.

[0396] Table 50

[0397]

[0398] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0399] Among the 353 pairs of donors and recipients in the HLA-matched cohort, 203 cases had GNRH2-KRT24 gene mutations, including 51 cases in the aGVHD-positive group and 152 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 51. After chi-square test analysis, P=0.0063, there was a statistically significant difference between the two groups.

[0400] Table 51

[0401]

[0402] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0403] 26. GNRH2-MAGEE2 gene mutation

[0404] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, there were 123 cases of GNRH2-MAGEE2 gene mutations, including 69 cases in the aGVHD-positive group and 54 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 52. After chi-square test analysis, P=0.0368, and there was a statistical difference between the two groups.

[0405] Table 52

[0406]

[0407] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0408] Among the 353 pairs of donors and recipients in the HLA-matched cohort, there were 220 cases of GNRH2-MAGEE2 gene mutations, including 55 cases in the aGVHD-positive group and 165 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 53. After chi-square test analysis, P=0.0032, there was a statistically significant difference between the two groups.

[0409] Table 53

[0410]

[0411] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0412] 27. GNRH2-OBSCN gene mutation

[0413] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, 67 cases had GNRH2-OBSCN gene mutations, including 44 cases in the aGVHD-positive group and 23 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 54. After chi-square test analysis, P=0.0019, there was a statistically significant difference between the two groups.

[0414] Table 54

[0415]

[0416] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0417] Among the 353 pairs of donors and recipients in the HLA-matched cohort, 126 cases had GNRH2-OBSCN gene mutations, including 33 cases in the aGVHD-positive group and 93 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 55. After chi-square test analysis, P=0.0338, and there was a statistically significant difference between the two groups.

[0418] Table 55

[0419]

[0420] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0421] 28. GPSM1-OBSCN gene mutation

[0422] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, 50 cases had GPSM1-OBSCN gene mutations, including 37 cases in the aGVHD-positive group and 13 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 56. After chi-square test analysis, P=0.0001, there was a statistically significant difference between the two groups.

[0423] Table 56

[0424]

[0425] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0426] Among the 353 pairs of donors and recipients in the HLA fully matched cohort, 96 cases had GPSM1-OBSCN gene mutations, including 27 cases in the aGVHD-positive group and 69 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 57. After chi-square test analysis, P=0.0217, there was a statistically significant difference between the two groups.

[0427] Table 57

[0428]

[0429] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0430] 29. HAPLN3-OBSCN gene mutation

[0431] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, 72 cases had HAPLN3-OBSCN gene mutations, including 44 cases in the aGVHD-positive group and 28 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 58. After chi-square test analysis, P=0.0203, there was a statistically significant difference between the two groups.

[0432] Table 58

[0433]

[0434] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0435] Among the 353 donor-recipient pairs in the HLA fully matched cohort, 138 cases had HAPLN3-OBSCN gene mutations, including 36 cases in the aGVHD-positive group and 102 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 59. After chi-square test analysis, P=0.0249, there was a statistically significant difference between the two groups.

[0436] Table 59

[0437]

[0438] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0439] 30. LCN10-CD33 gene mutation

[0440] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, there were 5 cases of LCN10-CD33 gene mutation, including 5 cases in the aGVHD-positive group and 0 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 60. After chi-square test analysis, P=0.0236, there was a statistical difference between the two groups.

[0441] Table 60

[0442]

[0443] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0444] Among the 353 donor-recipient pairs in the HLA fully matched cohort, there were 5 cases of LCN10-CD33 gene mutation, including 4 cases in the aGVHD positive group and 1 case in the aGVHD negative group; the aGVHD grouping and gene pair mutation status are shown in Table 61. After chi-square test analysis, P=0.0008, there was a statistically significant difference between the two groups.

[0445] Table 61

[0446]

[0447] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0448] 31. MS4A12-ACTN2 gene mutation

[0449] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, 19 cases had MS4A12-ACTN2 gene mutations, including 14 cases in the aGVHD-positive group and 5 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 62. After chi-square test analysis, P=0.0305, there was a statistically significant difference between the two groups.

[0450] Table 62

[0451]

[0452] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0453] Among the 353 donor-recipient pairs in the HLA fully matched cohort, there were 24 cases, including 9 cases in the aGVHD positive group and 15 cases in the aGVHD negative group; the aGVHD grouping and gene pair mutation status are shown in Table 63. After chi-square test analysis, P=0.0277, there was a statistically significant difference between the two groups.

[0454] Table 63

[0455]

[0456] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0457] 32. MS4A12-ZNF211 gene mutation

[0458] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, there were 10 cases of MS4A12-ZNF211 gene mutations, including 9 cases in the aGVHD-positive group and 1 case in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 64. After chi-square test analysis, P=0.0096, there was a statistically significant difference between the two groups.

[0459] Table 64

[0460]

[0461]

[0462] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0463] Among the 353 donor-recipient pairs in the HLA fully matched cohort, there were 11 cases, including 6 cases in the aGVHD positive group and 5 cases in the aGVHD negative group; the aGVHD grouping and gene pair mutation status are shown in Table 65. After chi-square test analysis, P=0.0038, there was a statistically significant difference between the two groups.

[0464] Table 65

[0465]

[0466] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0467] 33. RBMX-CD33 gene mutation

[0468] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, there were 4 cases of RBMX-CD33 gene mutation, including 4 cases in the aGVHD-positive group and 0 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 66. After chi-square test analysis, P=0.0435, there was a statistically significant difference between the two groups.

[0469] Table 66

[0470]

[0471] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0472] Among the 353 donor-recipient pairs in the HLA fully matched cohort, there were 6 cases, including 4 cases in the aGVHD positive group and 2 cases in the aGVHD negative group; the aGVHD grouping and gene pair mutation status are shown in Table 67. After chi-square test analysis, P=0.0041, there was a statistically significant difference between the two groups.

[0473] Table 67

[0474]

[0475]

[0476] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0477] 34. SIRPB1-ACTN2 gene mutation

[0478] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, 15 cases had SIRPB1-ACTN2 gene mutations, including 13 cases in the aGVHD-positive group and 2 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 68. After chi-square test analysis, P=0.0032, there was a statistically significant difference between the two groups.

[0479] Table 68

[0480]

[0481] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0482] Among the 353 donor-recipient pairs in the HLA fully matched cohort, 62 cases had SIRPB1-ACTN2 gene mutations, including 20 cases in the aGVHD-positive group and 42 cases in the aGVHD-negative group. The aGVHD grouping and gene pair mutation status are shown in Table 69. After chi-square test analysis, P=0.0086, indicating a statistically significant difference between the two groups.

[0483] Table 69

[0484]

[0485] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0486] 35. SIRPB1-OBSCN gene mutation

[0487] Among the 212 pairs of donors and recipients in the HLA haplotype-matched cohort, 41 cases had SIRPB1-OBSCN gene mutations, including 27 cases in the aGVHD-positive group and 14 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 70. After chi-square test analysis, P=0.0238, and there was a statistical difference between the two groups.

[0488] Table 70

[0489]

[0490] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0491] Among the 353 donor-recipient pairs in the HLA fully matched cohort, 160 cases had SIRPB1-OBSCN gene mutations, including 40 cases in the aGVHD-positive group and 120 cases in the aGVHD-negative group; the aGVHD grouping and gene pair mutation status are shown in Table 71. After chi-square test analysis, P=0.0370, and there was a statistically significant difference between the two groups.

[0492] Table 71

[0493]

[0494] Note: No mutation in the table means that at least one of the two genes in the gene pair has not mutated, and mutation means that both genes in the gene pair have mutated.

[0495] Example 2: Application of 35 LOF mutation gene pairs in predicting the risk of aGVHD after HLA haplotype-matched and / or HLA-matched alloHCT

[0496] 1. The 212 patients and their donors (212 pairs of donors and recipients) in Example 1 were used as research subjects. The cumulative scores of the LOF mutation gene pairs of the donors and recipients and the occurrence of aGVHD were tested by Mann-Whitney U. The results showed that the cumulative scores of the LOF mutation gene pairs in the aGVHD-positive group were significantly higher than those in the aGVHD-negative group ( Figure 1 , P<0.0001).

[0497] The receiver operating characteristic (ROC) curve was used to analyze the specificity and sensitivity of predicting aGVHD based on the cumulative score of LOF mutation gene pairs. The results showed that the area under the curve of the cumulative score of LOF mutation gene pairs for predicting aGVHD was 0.776 ( Figure 2, 95% confidence interval was 0.714-0.838), the cumulative score of the LOF mutation gene pair corresponding to the maximum Yoden Index (used to calculate the diagnostic cut-off value) was 1.5. Taking 1.5 as the diagnostic threshold for aGVHD, the detection sensitivity was 73.6% and the specificity was 71.7%.

[0498] 2. The 353 patients and their donors (353 pairs of donors and recipients) in Example 1 were used as research subjects. The cumulative scores of the LOF mutation gene pairs of the donors and recipients and the occurrence of aGVHD were tested by Mann-Whitney U. The results showed that the cumulative scores of the LOF mutation gene pairs in the aGVHD-positive group were significantly higher than those in the aGVHD-negative group ( Figure 3 , P<0.0001).

[0499] The receiver operating characteristic (ROC) curve was used to analyze the specificity and sensitivity of LOF mutation gene for predicting aGVHD. The results showed that the area under the curve of LOF mutation gene for predicting aGVHD was 0.729 ( Figure 4 , 95% confidence interval was 0.663-0.795), and the cumulative score of the LOF mutation gene pair corresponding to the maximum Yoden Index (used to calculate the diagnostic cut-off value) was 3.5. Taking 3.5 as the diagnostic threshold for aGVHD, the detection sensitivity was 53.5% and the specificity was 77.3%.

[0500] 3. All 565 patients and their donors (565 donor-recipient pairs) in Example 1 were used as research subjects. The cumulative scores of LOF mutation gene pairs of donors and recipients and the occurrence of aGVHD were tested by Mann-Whitney U. The results showed that the cumulative scores of LOF mutation gene pairs in the aGVHD-positive group were significantly higher than those in the aGVHD-negative group ( Figure 5 , P<0.0001).

[0501] The receiver operating characteristic (ROC) curve was used to analyze the specificity and sensitivity of LOF mutation gene for predicting aGVHD. The results showed that the area under the curve of LOF mutation gene for predicting aGVHD was 0.726 ( Figure 6 , 95% confidence interval was 0.682-0.770), the maximum Yoden Index (used to calculate the diagnostic cut-off value) corresponding to the gene pair mutation cumulative score was 1.5, and 1.5 was used as the diagnostic threshold for aGVHD. At this time, the sensitivity of the test was 74.6% and the specificity was 59.3%.

[0502] Therefore, in practical applications, the risk of aGVHD in patients with different modes of allogeneic hematopoietic transplantation can be predicted based on the cumulative scores of 35 LOF mutation gene pairs in donors and recipients. Then, pre-alloHCT donors can be screened or pre-alloHCT donors and recipients can be matched based on the risk of aGVHD, so as to optimize the selection of pre-alloHCT donors and improve the survival rate and quality of life of pre-alloHCT patients.

[0503] The cumulative score of the LOF mutation gene pair of each patient and their donor was obtained according to the following method (one patient was used as an example for explanation): the mutation of the protective mutation gene pair in the peripheral blood of the patient and their donor was detected to obtain the cumulative score of the protective mutation gene pair; the mutation of the promoting mutation gene pair in the peripheral blood of the patient and their donor was detected to obtain the cumulative score of the promoting mutation gene pair; the sum of the cumulative score of the protective mutation gene pair and the cumulative score of the promoting mutation gene pair was the cumulative score of the LOF mutation gene pair of the patient;

[0504] The protective mutant gene pairs are 19 LOF mutant gene pairs shown in Table 1 in Example 1 that are significantly negatively correlated with the occurrence of aGVHD (labeled as "aGVHD-" in Table 1), specifically as follows: ADAMTS7-APOL4, AGAP3-ACR, ANAPC1-ACR, CARD8-SIRPB1, DCAF15-ACR, DENND3-ACR, FRMD5-MYCT1, HAPLN3-ACR, HRNR-ACR, MAGEE2-ACR, MYCT1-KRTAP10-1, MYCT1-OAS1, NUDT1-ACR, OAS1-ACR, PABPC3-ACR, SIRPA-ACR, SLC35G6-ACR, TLR8-ACR, ZFPM1-ACR;

[0505] The promoting mutation gene pairs are 16 LOF mutation gene pairs shown in Table 1 in Example 1 that are significantly positively correlated with the occurrence of aGVHD (labeled as "aGVHD+" in Table 1), specifically as follows: ACTN2-OBSCN, CDH15-FRMD5, CDH15-GPR161, CDH15-OBSCN, GNRH2-FRMD5, GNRH2-KRT24, GNRH2-MAGEE2, GNRH2-OBSCN, GPSM1-OBSCN, HAPLN3-OBSCN, LCN10-CD33, MS4A12-ACTN2, MS4A12-ZNF211, RBMX-CD33, SIRPB1-ACTN2, SIRPB1-OBSCN;

[0506] The cumulative score of the protective mutation gene pairs is calculated as follows: if none of the 19 protective mutation gene pairs in the peripheral blood of the patient and the donor is mutated, it is recorded as 0; if one protective mutation gene pair (any one of the 19 protective mutation gene pairs) is mutated in the peripheral blood of the patient and the donor, it is recorded as -1; if two protective mutation gene pairs (any two of the 19 protective mutation gene pairs) are mutated in the peripheral blood of the patient and the donor, it is recorded as -2; and so on, if 19 protective mutation gene pairs are mutated in the peripheral blood of the patient and the donor, it is recorded as -19;

[0507] The calculation method for the cumulative score of the promoting mutation gene pairs is as follows: if none of the 16 promoting mutation gene pairs in the peripheral blood of the patient and its donor are mutated, it is recorded as 0; if one promoting mutation gene pair (any one of the 16 promoting mutation gene pairs) in the peripheral blood of the patient and its donor is mutated, it is recorded as 1; if two promoting mutation gene pairs (any two of the 16 promoting mutation gene pairs) in the peripheral blood of the patient and its donor are mutated, it is recorded as 2; and so on, if 16 promoting mutation gene pairs in the peripheral blood of the patient and its donor are mutated, it is recorded as 16.

[0508] The present invention has been described in detail above. It will be apparent to those skilled in the art that the present invention may be implemented in a wide range under equivalent parameters, concentrations and conditions without departing from the spirit and scope of the present invention and without the need for unnecessary experimentation. Although the present invention provides specific embodiments, it should be understood that further improvements may be made to the present invention. In short, according to the principles of the present invention, this application intends to include any changes, uses or improvements to the present invention, including changes made by conventional techniques known in the art that depart from the scope disclosed in this application. Applications of some of the basic features may be made within the scope of the following appended claims.

Claims

1. Use of a substance for detecting whether a gene pair in a donor and a recipient has mutated in any of the following A1)-A4): A1) Preparation of products for screening or assisting in the screening of donors prior to allogeneic hematopoietic cell transplantation; A2) Preparation of products for matching or assisting matching between donors and recipients before allogeneic hematopoietic cell transplantation; A3) preparing products for predicting or assisting in predicting the risk of acute graft-versus-host disease in patients after allogeneic hematopoietic cell transplantation; A4) Preparation of products for diagnosis or auxiliary diagnosis of acute graft-versus-host disease in patients after allogeneic hematopoietic cell transplantation; The gene pair is at least one of the following gene pairs: ADAMTS7-APOL4, AGAP3-ACR, ANAPC1-ACR, CARD8-SIRPB1, DCAF15-ACR, DENND3-ACR, FRMD5-MYCT1, HAPLN3-ACR, HRNR-ACR, MAGEE2-ACR, MYCT1-KRTAP10-1, MYCT1-OAS1, NUDT1-ACR, OAS1-ACR, PABPC3-ACR, SIRPA-ACR, SLC35G6-ACR, TLR 8-ACR, ZFPM1-ACR, ACTN2-OBSCN, CDH15-FRMD5, CDH15-GPR161, CDH15-OBSCN, GNRH2-FRMD5, GNRH2-KRT24, GNRH2-MAGEE2, GNRH 2-OBSCN, GPSM1-OBSCN, HAPLN3-OBSCN, LCN10-CD33, MS4A12-ACTN2, MS4A12-ZNF211, RBMX-CD33, SIRPB1-ACTN2, SIRPB1-OBSCN.

2. Use of a substance for detecting whether a gene pair in a donor and a recipient has mutated in any of the following B1)-B4): B1) Screening or assisting in the selection of donors before allogeneic hematopoietic cell transplantation; B2) matching or auxiliary matching between donor and recipient before allogeneic hematopoietic cell transplantation; B3) predict or assist in predicting the risk of acute graft-versus-host disease in patients after allogeneic hematopoietic cell transplantation; B4) To diagnose or assist in the diagnosis of acute graft-versus-host disease in patients after allogeneic hematopoietic cell transplantation; The gene pair is at least one of the following gene pairs: ADAMTS7-APOL4, AGAP3-ACR, ANAPC1-ACR, CARD8-SIRPB1, DCAF15-ACR, DENND3-ACR, FRMD5-MYCT1, HAPLN3-ACR, HRNR-ACR, MAGEE2-ACR, MYCT1-KRTAP10-1, MYCT1-OAS1, NUDT1-ACR, OAS1-ACR, PABPC3-ACR, SIRPA-ACR, SLC35G6-ACR, TLR 8-ACR, ZFPM1-ACR, ACTN2-OBSCN, CDH15-FRMD5, CDH15-GPR161, CDH15-OBSCN, GNRH2-FRMD5, GNRH2-KRT24, GNRH2-MAGEE2, GNRH 2-OBSCN, GPSM1-OBSCN, HAPLN3-OBSCN, LCN10-CD33, MS4A12-ACTN2, MS4A12-ZNF211, RBMX-CD33, SIRPB1-ACTN2, SIRPB1-OBSCN.

3. The use according to claim 1 or 2, characterized in that: The allogeneic hematopoietic cell transplantation includes HLA haplotype-matched allogeneic hematopoietic cell transplantation and HLA fully-matched allogeneic hematopoietic cell transplantation.

4. The use according to any one of claims 1 to 3, characterized in that: The substance for detecting whether a gene pair in a donor and a recipient has mutated is a substance for detecting whether a gene pair in the peripheral blood of a donor and a recipient has mutated.

5. The use according to any one of claims 1 to 4, characterized in that: The substance for detecting whether a gene pair in the peripheral blood of a donor and a recipient has mutated is a reagent and / or an instrument for detecting whether a gene pair in the peripheral blood of a donor and a recipient has mutated.

6. A kit having any one of the functions a1) to a4) below, comprising a substance for detecting whether a gene pair in a donor and a recipient has mutated; a1) Screening or assisting in the selection of donors before allogeneic hematopoietic cell transplantation; a2) Matching or auxiliary matching between donor and recipient before allogeneic hematopoietic cell transplantation; a3) To predict or assist in predicting the risk of acute graft-versus-host disease in patients after allogeneic hematopoietic cell transplantation; a4) To diagnose or assist in the diagnosis of acute graft-versus-host disease in patients after allogeneic hematopoietic cell transplantation; The gene pair is at least one of the following gene pairs: ADAMTS7-APOL4, AGAP3-ACR, ANAPC1-ACR, CARD8-SIRPB1, DCAF15-ACR, DENND3-ACR, FRMD5-MYCT1, HAPLN3-ACR, HRNR-ACR, MAGEE2-ACR, MYCT1-KRTAP10-1, MYCT1-OAS1, NUDT1-ACR, OAS1-ACR, PABPC3-ACR, SIRPA-ACR, SLC35G6-ACR, TLR 8-ACR, ZFPM1-ACR, ACTN2-OBSCN, CDH15-FRMD5, CDH15-GPR161, CDH15-OBSCN, GNRH2-FRMD5, GNRH2-KRT24, GNRH2-MAGEE2, GNRH 2-OBSCN, GPSM1-OBSCN, HAPLN3-OBSCN, LCN10-CD33, MS4A12-ACTN2, MS4A12-ZNF211, RBMX-CD33, SIRPB1-ACTN2, SIRPB1-OBSCN.

7. The kit according to claim 6, characterized in that: The allogeneic hematopoietic cell transplantation includes HLA haplotype-matched allogeneic hematopoietic cell transplantation and HLA fully-matched allogeneic hematopoietic cell transplantation.

8. The kit according to claim 6 or 7, characterized in that: The substance for detecting whether a gene pair in a donor and a recipient has mutated is a substance for detecting whether a gene pair in the peripheral blood of a donor and a recipient has mutated.

9. The kit according to any one of claims 6 to 8, characterized in that: The substance for detecting whether a gene pair in the peripheral blood of a donor and a recipient has mutated is a reagent and / or an instrument for detecting whether a gene pair in the peripheral blood of a donor and a recipient has mutated.

10. Use of ADAMTS7, APOL4, AGAP3, ACR, ANAPC1, CARD8, SIRPB1, DCAF15, DENND3, FRMD5, MYCT1, HAPLN3, HRNR, MAGEE2, KRTAP10-1, OAS1, NUDT1, PABPC3, SIRPA, SLC35G6, TLR8, ZFPM1, ACTN2, OBSCN, CDH15, GPR161, GNRH2, KRT24, GPSM1, LCN10, CD33, MS4A12, ZNF211 and / or RBMX as targets in the preparation of products for the prevention and / or treatment of acute graft-versus-host disease.

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