Complement-Induced Endothelial Cell Activation by a Novel Rab5-ZFYVE21-SMURF2 Signaling Axis

新型 Rab5-ZFYVE21-SMURF2 信号轴补体诱导内皮细胞激活

• 批准号: 10294356
• 负责人: DANIEL JANE-WIT
• 金额: $ 8.2万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10294356
• 关键词: Administrative Supplement; Adopted; Affect; Allografting; Antibodies; Appointment; Arteriosclerosis; Autoimmune; Award; Binding; Biopsy; Blood Component Removal; Cell Death; Cell surface; Clinic; Clinical; Collection; Complement; Complement Activation; Complement Membrane Attack Complex; Connective Tissue Diseases; Coronary artery; Diagnosis; Disease; Endosomes; Endothelial Cells; Enrollment; Ensure; Event; Experimental Models; Extramural Activities; Faculty; Failure; Fathers; Functional disorder; Funding; Histocompatibility Testing; Host Defense; Human; Human Resources; Immune; In Vitro; Individual; Inflammasome; Inflammatory; Infrastructure; International; Isoantibodies; Laboratories; Laboratory Finding; Lesion; Life; Logistics; Mediating; Mentors; Metastatic Pancreatic Adenocarcinoma; Microcirculatory Bed; Modeling; Molecular; Myocardial Infarction; Online Systems; Operative Surgical Procedures; Organ Transplantation; Output; PTEN gene; Pathologic; Pathology; Patients; Peripheral Blood Mononuclear Cell; Phase; Phenotype; Phosphotransferases; Population; Productivity; Protein Isoforms; Proteins; Protocols documentation; Publications; Publishing; Research; Resources; Sampling; Services; Signal Pathway; Signal Transduction; Solid; Specimen; Structure; Study models; Sum; System; Time; Training; Transplantation; Transplantation Surgery; Vesicle; Work; base; career; cell mediated immune response; clinically relevant; in vivo; lectures; membrane model; mortality; novel; phosphoinositide-3,4,5-triphosphate; primary caregiver; productivity loss; prospective; recruit; sample collection; symposium; transmission process; ubiquitin-protein ligase; vascular inflammation; virtual

PROJECT SUMMARY Transplant arteriosclerosis (TA) is a vaso-occlusive condition characterized by the formation of stenotic lesions in arterial and microvascular beds of solid organ allografts that cause ischemic graft loss. Complement (C’) are immune proteins involved in host defense that become pathologically activated on endothelial cells (ECs) during TA. In TA, complement becomes activated by recipient-derived alloantibodies binding to donor ECs. Upon activation complement proteins self-assemble to form membrane attack complexes (MAC), heterodimers that insert into target EC surfaces as transmembranous pores. The pathologic effects of complement activation including assembly on ECs is well established, however the precise molecular basis underlying these effects remains unclear, largely due to the lack of relevant experimental models for studying MAC. To obviate this, we developed models for antibody-mediated C’ activation on ECs in vitro and in vivo. Our protocols incorporated human biospecimens in order to recapitulate TA phenotypes in patients. Using these protocols, we defined a novel signaling mechanism causing EC activation and EC-mediated immune responses promoting TA. We have since built a collaborative infrastructure to obtain myriad patient biospecimens to support our work. These patient specimens include ‘high’ panel reactive antibody sera, fresh human coronary arteries, sera and PBMCs from prospectively enrolled patients, healthy PBMCs, TA biopsies, and biopsies from patients with various complement-mediated conditions involving vascular inflammation. Collection of these specimens involved collaborative interactions with the Tissue Typing Lab, Dept of Cardiothoracic Surgery, Dept of Transplant Surgery, Apheresis Service, and various Depts of Pathology. Due to the broad and extensive use of the materials above, my laboratory is exquisitely dependent on their ongoing collection for productivity. In this supplement, we propose a strategy to offset potential losses of productivity due to a critical life event, i.e., the diagnosis of metastatic pancreatic adenocarcinoma in my father. Supplemental funding will be used to hire a research technician who will possess the requisite skillsets and availability on nights and weekends to collect patient specimens. A plan for hiring, training, evaluating, and mentoring this individual is provided along with descriptions of current lab productivity, the critical life event, and a planned return to full productivity at the end of the supplemental period. Use of supplemental funding will occur contemporaneously with various mechanisms intended to maintain productivity both during the supplemental period and include institutional commitments for FMLA, benefits, protected time, colleague support, and wellness resources.

项目摘要 移植物动脉硬化（transplantarteriosclerosis，TA）是一种血管闭塞性疾病，其特征是狭窄血管的形成。 导致移植物缺血性丢失的实体器官移植物的动脉和微血管床病变。补充 （C '）是参与宿主防御的免疫蛋白，其在内皮细胞上被病理激活 (ECs)在TA期间。在TA中，补体被抗原衍生的同种抗体激活， EC。激活后，补体蛋白自组装形成膜攻击复合物（MAC）， 异二聚体作为跨膜孔插入靶EC表面。 补体激活包括组装对EC的病理作用已得到充分证实， 然而，这些效应背后的精确分子基础仍然不清楚，主要是由于缺乏 相关的实验模型进行MAC研究。为了证实这一点，我们开发了抗体介导的 体外和体内EC上的C'活化。我们的方案中加入了人类生物样本， 概括患者的TA表型。利用这些协议，我们定义了一种新的信令机制， 引起EC活化和EC介导的免疫应答促进TA。 此后，我们建立了一个协作基础设施，以获得无数的患者生物标本， 工作这些患者标本包括“高”组反应性抗体血清、新鲜人冠状动脉、血清 和来自前瞻性入组患者的PBMC、健康PBMC、TA活检组织和来自患有 涉及血管炎症的各种补体介导的病症。这些标本的采集 涉及与组织分型实验室，心胸外科， 移植手术、血液分离服务和病理学的各个部门。由于广泛和广泛的使用， 除了上述材料，我的实验室非常依赖于他们不断收集的生产力。 在本补充中，我们提出了一种策略，以抵消由于关键寿命而导致的潜在生产力损失 事件，即，我父亲被诊断为转移性胰腺癌补充资金将 过去常常雇用一名研究技术人员，他将拥有必要的技能和夜间可用性， 周末收集病人样本招聘、培训、评估和指导该员工的计划如下 提供沿着当前实验室生产率、关键生命事件和计划的全面恢复 在补充期结束时的生产力。补充资金的使用将同时进行 具有各种机制，旨在在补充期间保持生产力， 对FMLA、福利、受保护的时间、同事支持和健康资源的机构承诺。