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.

美国国立卫生研究院估计，2400万美国人患有自身免疫性疾病， 发生终末器官衰竭而器官移植是唯一治愈方法的人的百分比。 原发性自身免疫性疾病（RPAD）的复发是移植物失功的主要原因之一， 实体器官移植（SOT）后的发病率和死亡率。管理这些患者的主要挑战是 确定免疫介导的移植物功能障碍是自身免疫还是同种免疫过程的结果 或两者目前的诊断涉及血清学、放射学和组织学标准的组合， 明确区分急性细胞排斥反应（ACR）和RPAD。更好地理解 这两个过程之间的联系可能最终提高RPAD和ACR治疗的安全性和有效性， 针对个体患者移植物损伤的特定机制。适应性T和B细胞应答是 与自身免疫性疾病和同种异体移植排斥有关。我们建议使用一种新的方法来识别 同种异体反应性T细胞和自身反应性T和B细胞在移植后同种异体移植物功能障碍中的特殊作用 自身免疫性疾病患者。使用T细胞受体（TCR）β的高通量测序（HTS） 链互补决定区3（CDR 3），我们已经开发了一种新的方法， 追踪移植受者的同种异体反应我们现在建议结合使用这种方法 使用基于HTS的方法识别和跟踪自身免疫TCR和B细胞受体（BCR）库， 为了区分移植物排斥反应和RPAD在移植后移植物功能障碍中的作用， 自身免疫性疾病我们将使用肝移植（LT）治疗自身免疫性肝病（ALD）作为一个系统， 来测试这种方法。TCR β CDR 3（高变）区的HTS将用于鉴定供体- 移植前血液中的反应性T细胞克隆和来自患者肝外植体的自身反应性T和B细胞克隆， 活组织检查移植后，我们将在血液、移植物、胆汁和可能的粪便中追踪这些克隆， 以及在同种异体移植物功能障碍之后的时期，以便辨别供体特异性T细胞克隆的作用， 自身反应性T和B细胞引起移植物功能障碍，并评估其对治疗的反应性。我们 假设当移植物功能障碍时， 由复发性酒精性肝脏疾病引起，而供者反应性T细胞克隆将在ACR中占主导地位。我们还将测试 可以在粪便中识别和追踪这些淋巴细胞，如果阳性，最终可以评估为 这是一种非侵入性的方法，可以避免将来对肝脏活检的需要。如果成功，一个新的工具， 区分ACR和rALD将变得可用，从而更深入地了解肝功能障碍， LT和使改进的治疗。我们的新方法具有巨大的潜力，可以提供诊断和 SOT后同种免疫和自身免疫过程的预后生物标志物，并将适用于移植 在其他类型的SOT自身免疫性疾病的功能障碍。