TCR and BCR deep sequencing to distinguish autoimmune recurrence from allograft rejection

TCR 和 BCR 深度测序可区分自身免疫复发和同种异体移植排斥

• 批准号: 9753390
• 负责人: Megan Sykes
• 金额: $ 20.23万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9753390
• 关键词: Acute; Address; Affect; Allografting; American; Autoimmune Diseases; Autoimmune Hepatitis; Autoimmune Process; Autoimmune Responses; Autoimmunity; B-Lymphocyte Subsets; B-Lymphocytes; Bile fluid; Biological Markers; Biopsy; Biopsy Specimen; Blood; Blood Circulation; Blood specimen; Cells; Clinical; Clonal Expansion; Clone Cells; Complementarity Determining Regions; DNA; Diagnostic; Disease; Feces; Fingerprint; Functional disorder; Future; Heart; High-Throughput Nucleotide Sequencing; Histologic; Human; Immune; Immunoglobulin M; Immunophenotyping; Injury; Intestines; Kidney; Lead; Life; Liver; Liver Dysfunction; Liver diseases; Lung; Lymphocyte; Mediating; Memory B-Lymphocyte; Methods; Mononuclear; Morbidity - disease rate; Organ; Organ Transplantation; Organ failure; PTPRC gene; Patients; Peripheral Blood Mononuclear Cell; Precision therapeutics; Process; Prognostic Marker; Radiology Specialty; Receptor Cell; Receptors, Antigen, B-Cell; Recurrence; Role; Safety; Serological; Solid; Stains; System; T-Cell Receptor; T-Lymphocyte; Testing; Therapeutic Effect; Therapeutic Intervention; Tissues; Transplant Recipients; Transplant-Related Disorder; Transplantation; United States National Institutes of Health; Urine; V(D)J Recombination; allograft rejection; autoreactive B cell; autoreactive T cell; autoreactivity; base; beta Chain Antigen T Cell Receptor; complementarity-determining region 3; curative treatments; deep sequencing; diagnostic biomarker; improved; individual patient; islet; kidney allograft; liver biopsy; liver transplantation; mortality; novel; novel strategies; peripheral blood; post-transplant; primary sclerosing cholangitis; response; stool sample; success; tool

Project Summary: NIH estimates that 24 million Americans suffer from autoimmune diseases, a significant percentage of whom develop end organ failure for which organ transplant would be the only curative treatment. Recurrence of primary autoimmune diseases (RPAD) is among the leading causes of graft loss and patient morbidity and mortality following solid organ transplantation (SOT). A major challenge in managing these patients is determining whether immune-mediated graft dysfunction is the result of an autoimmune or alloimmune process or both. Current diagnostics involving a combination of serological, radiological and histological criteria do not clearly distinguish between acute cellular rejection (ACR) and RPAD. A better understanding of the interplay between these two processes may ultimately improve the safety and efficacy of RPAD and ACR treatment, targeting the specific mechanism of graft injury in individual patients. Adaptive T and B cell responses are implicated in both autoimmune disorders and allograft rejection. We propose to use a novel approach to identify the specific roles of alloreactive T cells and autoreactive T and B cells in post-transplant allograft dysfunction in patients with autoimmune disease. Using high-throughput sequencing (HTS) of the T-cell receptor (TCR) beta chain complementarity-determining region 3 (CDR3), we have developed a novel method of identifying and tracking the human alloresponse in transplant recipients. We now propose to use this method in combination with a HTS-based method of identifying and tracking the autoimmune TCR and B-cell receptor (BCR) repertoire to distinguish the roles of allograft rejection and RPAD in graft dysfunction following transplantation due to autoimmune disease. We will use liver transplantation (LT) for autoimmune liver diseases (ALD) as a system in which to test this approach. HTS of the TCR beta CDR3 (hypervariable) region will be used to identify donor- reactive T cell clones in pre-transplant blood and autoreactive T and B cell clones from patient liver explants and biopsies. Following the transplant, we will track these clones in the blood, graft, bile and possibly stool during and following periods of allograft dysfunction in order to discern the roles of donor-specific T cell clones and autoreactive T and B cells in causing graft dysfunction and to assess their responsiveness to therapy. We hypothesize that autoreactive clones will be more abundant in blood or liver tissues when graft dysfunction is caused by recurrent ALD, whereas donor-reactive T cell clones will predominate in ACR. We will also test whether it is possible to identify and track these lymphocytes in stool, which, if positive, could ultimately be evaluated as a non-invasive method that would avoid the need for liver biopsies in the future. If successful, a new tool to distinguish ACR and rALD will become available, allowing a deeper understanding of liver dysfunction following LT and enabling improved therapies. Our novel approach has enormous potential to provide a diagnostic and prognostic biomarker of alloimmune and autoimmune processes following SOT and will be applicable to graft dysfunction in other types of SOT for autoimmune diseases.

项目摘要：NIH估计有2400万美国人患有自身免疫性疾病，这是一个重大的 其中有器官移植是唯一的治疗方法的最终器官衰竭的百分比。 原发性自身免疫性疾病（RPAD）的复发是移植和患者的主要原因之一 固体器官移植（SOT）后的发病率和死亡率。管理这些患者的主要挑战 正在确定免疫介导的移植功能障碍是自身免疫性或同种免疫过程的结果 或两者兼而有之。当前涉及血清学，放射学和组织学标准组合的诊断不 明确区分急性细胞排斥（ACR）和RPAD。更好地理解相互作用 在这两个过程之间可能最终提高RPAD和ACR治疗的安全性和功效， 针对个别患者的移植物损伤的特定机制。自适应T和B细胞反应是 与自身免疫性疾病和同种异体移植排斥相关。我们建议使用一种新颖的方法来识别 同种异体T细胞以及自动反应性T和B细胞在移植后同种异体功能障碍中的特定作用 自身免疫性疾病的患者。使用T细胞受体（TCR）β的高通量测序（HTS） 链互补性确定的区域3（CDR3），我们开发了一种新颖的方法来识别和 跟踪移植受体中的人类同种异腺响应。我们现在建议将此方法组合使用 采用基于HTS的方法来识别和跟踪自身免疫性TCR和B细胞受体（BCR）曲目 为了区分同种异移植排斥和RPAD在移植后移植功能障碍中的作用 自身免疫性疾病。我们将使用肝移植（LT）进行自身免疫性肝病（ALD）作为系统 要测试这种方法。 TCR Beta CDR3（高变量）区域的HTS将用于识别供体 病房前血液中的反应性T细胞克隆，自动反应性T和B细胞克隆来自患者肝外植体，以及 活检。移植后，我们将在血液，移植物，胆汁和可能的粪便中跟踪这些克隆 以及以下同种异体功能障碍的时期，以辨别供体特异性T细胞克隆和 自动反应性T和B细胞引起移植功能障碍并评估其对治疗的反应。我们 假设自动反应性克隆在血液或肝组织中会更丰富 由复发性ALD引起的，而供体反应性T细胞克隆将在ACR中占主导地位。我们还将测试是否 可以在粪便中识别和跟踪这些淋巴细胞，如果阳性，最终可以评估为 一种非侵入性方法，它将避免将来需要进行肝活检。如果成功，那是一个新工具 区分ACR和Rald将可以使用，从而更深入地了解肝功能障碍 LT并启用改进的疗法。我们的新方法具有提供诊断和的巨大潜力 SOT之后的Allomune和自身免疫过程的预后生物标志物，将适用于移植物 自身免疫性疾病的其他类型的SOT功能障碍。