Graft extracellular vesicles as promoters of anti-donor immunity in cardiac and skin transplantation

移植细胞外囊泡作为心脏和皮肤移植中抗供体免疫的促进剂

• 批准号: 10560271
• 负责人: Adrian E. Morelli
• 金额: $ 65.59万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-19 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10560271
• 关键词: Adult; Affect; Allogenic; Allografting; Antigen-Presenting Cells; Antigens; Apoptotic; B-Lymphocytes; Biogenesis; Biological Markers; Biology; Blood group antigen S; Cardiac; Cardiovascular system; Caring; Cells; Childhood; Chronic; Clinical; Communication; Development; Disease; Disease Marker; Distant; Electron Microscopy; Experimental Models; Family; Future; Generations; Genetically Engineered Mouse; Goals; Graft Rejection; Heart; Heart Transplantation; Heart failure; Human; Immune; Immune response; Immune system; Immunity; Immunosuppression; Immunosuppressive Agents; In Situ; Intervention; Investigation; Knowledge; Lead; Leukocytes; Lipids; Lung; Lymphoid Tissue; MHC antigen; Malignant Neoplasms; Mediating; Mediator of activation protein; Medical; Messenger RNA; Methodology; Mus; Organ; Patients; Peptides; Proteins; Regimen; Resolution; Risk; Role; Secondary to; Signal Transduction; Skin; Skin Transplantation; Structure of germinal center of lymph node; T memory cell; T-Lymphocyte; Testing; Therapeutic immunosuppression; Tissue Grafts; Tissue Transplantation; Transplantation; Travel; Untranslated RNA; Vesicle; adaptive immune response; allograft rejection; base; bioimaging; clinically relevant; conditioning; exosome; extracellular vesicles; heart allograft; horizontal cell; humanized mouse; immunoreaction; immunoregulation; improved; in vivo; infection risk; isoimmunity; microvesicles; mouse model; novel; novel therapeutic intervention; prevent; promoter; response; side effect; skin allograft; translational model; transplant model; two photon microscopy; vesicular release

SUMMARY The innate and adaptive immune response against allografts is the main impediment to successful transplantation. Heart transplantation is the best option for selected pediatric and adult patients with end-stage heart failure, however, the threat of rejection remains high, despite improvements in immunosuppressive therapies, conditioning regimens and medical care. Importantly, currently used interventions are not donor-specific and therefore may cause increased risk of infections and cancer. A deeper understanding of the basis of allo-recognition is needed for the development of novel donor-specific therapies to treat graft rejection minimizing the harmful side effects. Recent studies have challenged the dogma that initiation or re-activation of anti-donor immunity in graft- draining secondary lymphoid tissues (SLTs) depends mainly on donor Ag-presenting cells (APCs) mobilized from the grafts. Increasing evidence and our preliminary studies indicate that heart and non-vascularized skin allografts release extracellular vesicles (EVs) carrying donor-Ag that traffic to the SLTs where the graft EVs stimulate donor- reactive B cells and T cells. Despite increasing information during the past 5 years on the role of graft EVs on elicitation of anti-donor immunity and their potential use as biomarkers in transplantation, the mechanisms in vivo by which graft EVs interact with recipient’s immune cells in graft-draining SLTs and promote anti-donor immunity remain largely unknown. The family of EVs encompasses vesicles with different biogenesis, size and composition that includes exosomes and microvesicles. Although growing evidence indicates that EVs represent a mechanism by which cells horizontally transfer proteins, mRNAs, non-coding RNAs and lipids, the function of EVs in vivo remains an enigma. Therefore, we propose to investigate the mechanisms in vivo by which graft EVs initiate or re-activate in graft-draining SLTs, the innate and adaptive immune responses that lead to rejection of allografts. We will analyze these mechanisms in mouse experimental models of cardiac and non-vascularized skin allografts. We hypothesize that “graft EVs constitute a cell-free platform that by multiple mechanisms initiates or re-activates the anti-donor immune response in the recipient’s SLTs”. This application will investigate these mechanisms in situ and in vivo using transplant models in mice and a translational model in humanized mice. We will test our hypothesis in the following aims: Aim 1 will investigate the origin of graft EVs and their effects on recipient APCs in graft-draining SLTs, Aim 2 will analyze the mechanisms by which graft EVs generate anti-donor B cell immunity in SLTs and Aim 3 will analyze how graft EVs elicit anti-donor T cell immunity in SLTs and its relevance to transplantation in humans. Our long-term goal is to understand how graft EVs function in vivo to provide new grounds for development of EV-based therapies and disease markers of clinical relevance to transplantation and immune-mediated disorders.

概括 对同种异体移植的先天和适应性免疫响应是成功移植的主要障碍。 心脏移植是选定的儿科和成年患者终末期心力衰竭的最佳选择， 但是，拒绝的威胁仍然很高，免疫抑制疗法的目的地改善，调理 方案和医疗保健。重要的是，目前使用的干预措施不是捐助者特定的，因此可能 导致感染和癌症的风险增加。需要更深入地理解Allo-Revition的基础 为了开发新颖的捐助者特异性疗法，以治疗等级排斥，最大程度地减少有害副作用。 最近的研究挑战了教条，即在移植物中主动或重新激活抗唐纳免疫力 排水次级淋巴组织（SLT）主要取决于供体的呈递细胞（APC） 移植物。越来越多的证据和我们的初步研究表明，心脏和非血管化皮肤外观 释放带有供体的供体的细胞外蔬菜（EVS），该供体流量到SLTS刺激供体的供体 - 反应性B细胞和T细胞。尽管在过去的5年中越来越多的信息就移植电动汽车的作用 抗抑制免疫及其作为移植生物标志物的潜在用途，体内机制 接枝EVS与受体排水SLT中的免疫球相互作用并促进抗抑制免疫 在很大程度上未知。 电动汽车家族包括具有不同的生物发生，大小和成分的蔬菜，其中包括 外泌体和微囊泡。虽然越来越多的证据表明EV代表了一种机制 细胞水平转移的蛋白质，mRNA，非编码RNA和脂质，电动汽车在体内的功能仍然是 谜。因此，我们建议研究接枝EV启动或重新激活的体内机制 在流血的SLT中，导致同种异体移植的固有和适应性免疫反应。我们将 在心脏和非血管化皮肤异源的小鼠实验模型中分析这些机制。 我们假设“移植电动汽车构成一个通过多种机制启动的无细胞的平台 或重新激活接收者SLT中的抗抑制免疫反应”。该应用程序将调查 这些机制原位和体内使用小鼠中的移植模型和人源化的翻译模型 老鼠。我们将在以下目的中检验我们的假设：AIM 1将研究移植电动汽车及其的起源 AIM 2对接枝排水SLT中的受体APC的影响将分析移植EVS产生的机制 SLTS中的抗Donor B细胞免疫史和AIM 3将分析移植EVS如何引起抗Donor T细胞免疫历史 及其与人类移植的相关性。我们的长期目标是了解Graft EV在 体内为开发基于EV的疗法和临床相关性的疾病标志物提供新的基础 移植和免疫介导的疾病。