The NOTCH Signaling Pathway in Large Vessel Vasculitis

大血管炎中的 NOTCH 信号通路

• 批准号: 8629407
• 负责人: Cornelia M. Weyand
• 金额: $ 41.57万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-06 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8629407
• 关键词: 1-Phosphatidylinositol 3-Kinase; 3-Dimensional; Acute T Cell Leukemia; Adhesiveness; Adrenal Cortex Hormones; Adult; Affect; Antibodies; Antigens; Aorta; Aortic Aneurysm; Aortic Arch Syndromes; Aortic Diseases; Arteries; Arteritis; Automobile Driving; Behavior; Biological Models; Biomedical Engineering; Blindness; Blood Vessels; CCL2 gene; CD4 Positive T Lymphocytes; Calcineurin; Cell Communication; Cell Differentiation process; Cell Survival; Cell physiology; Cells; Cellulose; Chimera organism; Chimeric Proteins; Chronic; Clinic; Clinical; Clonal Expansion; Communication; Custom; Dendritic Cells; Development; Disease; Dose; Endothelial Cells; Fiber; Growth; Human; Hyperplasia; Hypertension; Immune; Immune response; Immune system; Immunity; Indium; Inflammation; Inflammatory; Interferons; Interleukin-17; Knowledge; Lesion; Life; Ligands; Mediating; Modeling; Molecular; Myocardial; NOTCH1 gene; Oncogenic; Pathway interactions; Patients; Pattern; Phenotype; Point Mutation; Population; Process; Production; RNA Interference; Resources; Role; SCID Mice; Series; Shapes; Signal Pathway; Signal Transduction; Smooth Muscle Myocytes; Stroke; System; T cell response; T-Cell Proliferation; T-Lymphocyte; TCF3 gene; Takayasu' s Arteritis; Technology; Temporal Arteritis; Testing; Therapeutic; Tissues; Transplantation; Tumor Suppressor Proteins; Vascular Endothelial Cell; Vasculitis; aging population; base; body system; c-myc Genes; cell behavior; cell growth; cohort; cytokine; design; human FRAP1 protein; in vivo; inhibitor/antagonist; interleukin-22; migration; mouse model; notch protein; novel; novel therapeutic intervention; novel therapeutics; overexpression; public health relevance; receptor; receptor expression; response; restoration; scaffold; small molecule

Project Summary Large vessel vasculitides (LVV), such as giant cell arteritis (GCA) cause blindness, stroke, aortic arch syndrome, aortic aneurysm, hypertension and myocardial insufficiency. In an aging population the number of patients requiring chronic management for LVV has been steadily rising, while the therapeutic armamentarium has remained strictly limited to high-dose corticosteroids. The last decade has seen exciting progress in implicating the innate and adaptive immune system in the immunopathogenesis of LVV. However, there is a critical gap in our knowledge why the disease targets the aorta and its major branches and how immuno- stromal communications in the arterial wall initiate and promote vasculitis. The pathogenic immune response has a signature of antigen-induced clonal expansion, but we have recently seen that costimulatory signals deriving from resident cells in the tissue niche are equally important in driving tissue-damaging immunity. GCA arteries express abundant levels of NOTCH receptors and ligands, providing a molecular platform for superb cell-to-cell communication. Blocking of NOTCH signaling effectively inhibits vasculitis. CD4 T cells from GCA patients constitutively express NOTCH1 receptor, enabling them to interact with NOTCH ligand expressing vascular smooth muscle cells (VSMC) and endothelial cells (EC). This application is designed to uncover how the Notch pathway participates in immuno-endothelial and immuno-stromal communications and how NOTCH- dependent signaling shapes vasculitogenic T cell responses and maladaptive VSMC and EC behavior. The project builds on a series of enabling resources; including a clinically phenotyped cohort of GCA patients; a novel 3-D model system of human arterial walls which permits assembly of custom-made vessels from stackable units populated with defined cell populations; and a humanized mouse model carrying inflamed human arteries. Access to Notch receptor and ligands can be blocked through ligand-competing antibodies/fusion proteins and cells can be rendered Notch signaling deficient by RNAi technology. Specific Aim 1 examines on a mechanistic level how NOTCH ligands on VSMC and EC regulate effector functions of vasculitogenic CD4 T cells; modulate their growth, tissue invasion capacity and cytokine production. Specific Aim 2 seeks to identify signaling networks that can be utilized to either suppress NOTCH1 expression or target NOTCH-dependent survival signals in pathogenic T cells. Small molecule inhibitors disrupting Notch-derived signals will be tested in the chimera model for their anti-vasculitic potential. Specific Aim 3 is focused on the role of VSMC as signal-sending and signal-receiving cells and determines how NOTCH-NOTCH ligand interactions affect VSMC survival, migration, matrix production, contractility and ROS release. Specific Aim 4 unravels the molecular mechanisms through which patient-derived CD4+NOTCH1+ T cells regulate the functional behavior of ECs and investigates how such T cells modulate EC proinflammatory functions, angiogenic capacity, adhesiveness and leakiness of the EC barrier.

项目摘要 大血管血管（LVV），例如巨细胞炎（GCA）导致失明，中风，主动脉弓 综合征，主动脉瘤，高血压和心肌功能不全。在老龄化的人口中 需要长期管理LVV的患者一直在稳步上升，而治疗性武器库 严格仅限于大剂量皮质类固醇。过去十年在 将先天和适应性免疫系统牵涉到LVV的免疫病变中。但是，有一个 在我们的知识上，关键的差距为什么疾病针对主动脉及其主要分支以及如何免疫 动脉壁中的基质通信启动并促进血管炎。致病性免疫反应 具有抗原引起的克隆膨胀的签名 从组织生态裂市场中衍生的居民细胞在驱动组织受损的免疫力方面同样重要。 GCA 动脉表达大量的缺口受体和配体，为出色提供了一个分子平台 细胞间通信。凹槽信号的阻塞有效地抑制了血管炎。 GCA的CD4 T细胞 患者组成性表达Notch1受体，使他们能够与表达的Notch配体相互作用 血管平滑肌细胞（VSMC）和内皮细胞（EC）。该应用程序旨在发现如何 Notch途径参与免疫 - 内皮和免疫质通信，以及Notch- 依赖的信号传导塑造血管生成T细胞反应以及适应不良的VSMC和EC行为。这 项目建立在一系列启用资源的基础上；包括临床表型GCA患者的队列；一个 人类动脉壁的新型3-D模型系统，允许从 可堆叠的单元，该单元被定义的细胞群体；以及带有发炎的人源化鼠标模型 人动脉。可以通过配体竞争来阻止使用Notch受体和配体 RNAi技术缺乏缺陷的抗体/融合蛋白和细胞。具体的 AIM 1检查机械级别 血管生成CD4 T细胞；调节其生长，组织侵袭能力和细胞因子产生。具体的 AIM 2试图识别可用于抑制Notch1表达或目标的信号网络 病原T细胞中的Notch依赖性生存信号。小分子抑制剂破坏了缺口衍生的 信号将在Chimera模型中测试其抗血脉脉络膜电位。具体目标3专注于 VSMC作为信号端端和信号接收单元的作用，并确定Notch-Notch配体如何 相互作用会影响VSMC生存，迁移，基质产生，收缩力和ROS释放。具体目标4 解散患者衍生的CD4+ Notch1+ T细胞的分子机制 EC的功能行为，并研究了此类T细胞如何调节EC促炎函数， EC屏障的血管生成能力，粘附性和渗漏。