Identification of Canine Minor Histocompatibility Antigens

犬次要组织相容性抗原的鉴定

• 批准号: 8067930
• 负责人: Jay Ashok Shendure
• 金额: $ 36.51万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8067930
• 关键词: Acute leukemia; Address; Age; Alleles; Allogenic; Animals; Antigens; Biological Assay; Breeding; Canis familiaris; Cell Transplantation; Cells; Chimerism; Clinic; Code; Collaborations; Comorbidity; Custom; Cyclosporine; Cyclosporins; Data; Development; Donor Lymphocyte Infusion; Experimental Models; Functional RNA; Genes; Genetic; Genetic Polymorphism; Genetic Variation; Genomics; Genotype; Goals; Graft vs Tumor Effect; Harvest; Hematologic Neoplasms; Hematopoiesis; Hematopoietic; Histocompatibility Antigens; Human; Immunization; Immunosuppression; Immunosuppressive Agents; Injection of therapeutic agent; Knowledge; Lymphocyte; Malignant Neoplasms; Methodology; Minor; Minor Histocompatibility Antigens; Minority; Modeling; Molecular; Outcome; Patients; Peptides; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Price; Principal Investigator; Procedures; Proteins; Public Health; Reaction; Regimen; Relapse; Residual state; Reverse Transcriptase Polymerase Chain Reaction; Risk; Role; Shotgun Sequencing; Single Nucleotide Polymorphism; T cell response; T-Lymphocyte; Technology; Testing; Tissues; Translating; Transplant Recipients; Transplantation; Variant; base; conditioning; cost; design; experience; graft vs host disease; graft vs leukemia effect; high risk; high throughput screening; improved; in vivo; mycophenolate mofetil; next generation; novel; novel strategies; peripheral blood; pre-clinical; prevent; programs; pup; response; tumor

PROJECT 2: IDENTIFICATION OFCANINE MINOR HISTOCOMPATIBILITY ANTIGENS Project 1 has developed an approach at DLA-identical canine hematopoietic cell transplantation (HCT) that results in stable mixed donor-host chimerism. Persistent host hematopoiesis can serve as an experimental model of persistent hematologic malignancy seen in some patients transplanted under Projects 3 and 4. Conversion of mixed to all-donor chimerism can be achieved with injection of donor lymphocytes that have been sensitized to host minor histocompatibility antigens expressed on peripheral blood mononuclear cells (PBMC), however at the price of often fatal graft-vs.-host disease (GVHD). T-cell responses directed against ubiquitously expressed minor antigens are thought to be responsible for GVHD, while T-cell responses against a combination of ubiquitous and hematopoietic-specific minor antigens contribute to elimination of residual host hematopoietic cells in a manner analogous to the graft-vs.-leukemia effect observed in human patients. The identification of minor antigens restricted to hematopoietic cells therefore holds great promise for improving allogeneic HCT outcomes. That knowledge would facilitate the development of sensitization strategies that target host hematopoietic cells while sparing GVHD target tissues. However, while the dog model of allogeneic HCT is optimal for preclinical development of novel HCT therapies, no canine minor, histocompatibility antigens have been described to date, and existing methodologies for minor antigen identification are inefficient. To address this problem, we propose a novel approach to minor antigen discovery in the dog. This approach utilizes next generation sequencing technology to define protein coding variations unique to the recipient and expressed in PBMC which will be used for sensitizing the HCT donor. Sensitized donor T-cells will then be injected into the respective recipients with the aim of converting mixed to full-donor chimerism and causing GVHD. After conversion has been accomplished, T-cells will be harvested from recipients and tested for responses against candidate minor antigens using a novel, high- throughput T-cell assay. Positive responses would define genuine minor histocompatibility antigens. Using qRT-PCR, we will then identify those minor antigens that are highly expressed in hematopoietic cells but not in GVHD target tissues. Next, relevant minor antigen peptides will be used to sensitize donor T-cells with the aim of converting mixed to full-donor chimerism without GVHD (Project 1). Eventually, this concept will be tested in a canine model of acute leukemia in Project 1. Benefits to Public Health: Taken together, the studies proposed in this Project and the in vivo studies proposed in Project 1 have the potential of developing new and effective approaches benefiting patients with persisting/relapsing malignancies treated by allogeneic HCT under Projects 3 and 4.

项目2：识别少数组织相容性抗原 项目1已开发了一种在DLA-认同的犬造血细胞移植（HCT）的方法 导致稳定的混合供体宿主嵌合。持续的宿主造血可以用作实验 在项目3和4下移植的一些患者中，在一些患者中看到持续性血液系统恶性肿瘤的模型。 通过注射具有 被敏化以寄主在外周血单核细胞上表达的次要组织相容性抗原 （PBMC），但是以通常致命的移植物为宿主病（GVHD）的价格。针对的T细胞回答 普遍表达的次要抗原被认为是GVHD的原因，而T细胞反应 与普遍存在和造血特异性次要抗原的结合有助于消除 残留的宿主造血细胞的方式类似于移植物-V。 患者。因此，仅限于造血细胞的次要抗原鉴定有很大的希望 用于改善同种异体HCT结果。这些知识将促进敏化的发展 靶向宿主造血细胞的策略，同时保留GVHD靶组织。但是，狗 同种异体HCT的模型对于新型HCT疗法的临床前开发是最佳的，没有犬小犬， 迄今为止已经描述了组织相容性抗原和次要抗原的现有方法论 识别效率低下。为了解决这个问题，我们提出了一种新型抗原的新方法 在狗中发现。这种方法利用下一代测序技术定义蛋白质编码 接收者独有的变化并在PBMC中表达，该变化将用于使HCT供体敏化。 然后将敏化的供体T细胞注入各自的接受者，目的是转化混合 为了全米嵌合体并引起GVHD。转换完成后，T细胞将是 从接受者那里收获，并使用一种新型高级，对候选小抗原的反应进行了测试 吞吐量T细胞测定。积极的反应将定义真正的较小的组织兼容性抗原。使用 QRT-PCR，我们将识别那些在造血细胞中高度表达的次要抗原，但不能 在GVHD目标组织中。接下来，相关的次要抗原肽将用于使供体T细胞与 在没有GVHD的情况下，将混合变成全盘嵌合的目的（项目1）。最终，这个概念将是 在项目1中的急性白血病犬模型中进行了测试。公共卫生的好处： 该项目提出的研究和项目1中提出的体内研究具有 开发新的有效方法，使受过持续/复发性恶性肿瘤的患者受益 根据项目3和4的同种异体HCT。