Deciphering mechanisms of vascular disease with cell and process specific ligands

用细胞和过程特异性配体破译血管疾病的机制

基本信息

批准号：
10368129
负责人：
William H Thiel
金额：
$ 38.13万
依托单位：
UNIVERSITY OF IOWA
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-06-15 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10368129
关键词：
Acute Address Agonist Apoptosis Apoptotic Arterial Disorder Arterial Injury Binding Biological Assay Blood Vessels Blood flow Bypass CRISPR library Cell Proliferation Cell Surface Proteins Cell physiology Cell surface Cells Cessation of life Data Disease Endothelial Cells Endothelium Event Fluorescence Goals Growth Inhibitors Impairment In Vitro Injury Ligands Mediating Occlusion injury Operative Surgical Procedures Oranges PDGFRB gene Pathologic Pharmaceutical Preparations Phenotype Platelet-Derived Growth Factor Platelet-Derived Growth Factor beta Receptor Procedures Process Proliferating Reporting Risk Safety Signal Pathway Signal Transduction Smooth Muscle Myocytes Specificity Stents Testing Tissues Vascular Diseases Vascular Smooth Muscle Vein graft Work aptamer attenuation base cell dedifferentiation cell growth cell motility cell type common treatment drug efficacy efficacious treatment healing heart allograft improved in vivo in vivo Model innovation migration novel platelet-derived growth factor BB preservation prevent public health relevance response restenosis stent thrombosis targeted agent targeted treatment tool vascular abnormality vascular injury vascular smooth muscle cell migration vascular smooth muscle cell proliferation whole genome

项目摘要

Project Summary / Abstract The primary mechanism responsible for many acute vascular disorders, including in-stent restenosis, vein graft disease, and cardiac allograft arteriopathy, is pathological vascular smooth muscle cell (VSMC) activation. VSMC activation leads to neointimal formation and re-occlusion of injured blood vessels. Strategies to prevent abnormal VSMC remodeling, such as cell growth inhibitors used in drug eluting stents, have the undesired effect of impairing re-endothelialization. This attenuation of endothelial healing increases the risk of neoatherosclerosis and late stent thrombosis. In order to prevent pathological VSMC growth, but not re- endothelialization, there is a critical need to identify cell-surface proteins that may can be targeted by agents to distinguish between VSMCs and endothelial cells (ECs) and modulate cellular processes utilized by VSMCs during pathological remodeling that do not affect endothelium healing. The objective of this proposal is to utilize VSMC-targeting aptamers that specifically modulate VSMC migration, proliferation and apoptosis to define 1) define the cell-surface proteins and mechanism of action by which VSMC-targeting aptamers modulate VSMC but not EC processes; and 2) determine the impact of these cell- and process-specific ligands on neointimal formation and re-endothelialization following acute vascular injury. We recently identified a VSMC-specific anti- migratory aptamer that prevents VSMC migration with no effect on EC migration. We determined that the VSMC anti-migratory aptamer operates by antagonizing PDGFR-β activation, but has no effect on PDGF-BB- mediated VSMC proliferation. These data suggest the novel concept that PDGFR-β migration and proliferation signaling may be dissociated. We have now identified two additional VSMC-targeting aptamers that modulate VSMC, but not EC, proliferation and apoptosis through unknown mechanisms originating at the cell surface. We will test the overall hypothesis that VSMC cell surface proteins and signaling pathways necessary for migration, proliferation and apoptosis following acute vascular injury may be cell-specifically modulated to prevent neointimal formation without altering EC re-endothelialization. In Aim 1, we will define the mechanism by which VSMC PDGFR-β dependent migration is dissociated from PDGFR-β dependent proliferation and may be inhibited to prevent neointimal formation without interfering with re-endothelialization. Aim 2 will identify the cell-surface protein, using a novel application of a whole-genome CRISPR library, and define the mechanisms where VSMC proliferation may be specifically inhibited in VSMCs to prevent neointimal formation without impacting re-endothelialization. Aim 3 will determine the mechanism and cell surface-proteins by which VSMC apoptosis may be induced to limit neointimal formation while preserving re-endothelialization. Completion of this study will result in a mechanistic understanding of modulating VSMC migration, proliferation and apoptosis towards preventing neointimal formation and preserving re-endothelialization, with applicability to a number of acute vascular diseases.

项目摘要 /摘要负责许多急性血管疾病的主要机制，包括静脉内病，静脉移植疾病和心脏同类动脉炎是病理血管平滑肌细胞（VSMC）激活。 VSMC激活导致受伤的血管的新内膜形成和重纹。预防策略异常的VSMC重塑，例如药物洗脱支架中使用的细胞生长抑制剂，具有不希望的损害重新皮层化的影响。内皮愈合的这种衰减增加了 Neoatherssclosis和晚支架血栓形成。为了防止病理VSMC的增长，但不能内皮化，至关重要的是识别药物可能针对的细胞表面蛋白在VSMC和内皮细胞（EC）之间与众不同，并调节VSMC使用的细胞过程在病理重塑期间，不会影响内皮愈合。该提议的目的是利用特异性调节VSMC迁移，增殖和凋亡的VSMC靶向适体以定义1）定义细胞表面蛋白和作用机理，通过该蛋白调节VSMC靶向Aptamers调节VSMC 但不是EC过程； 2）确定这些细胞和过程特异性配体对新源的影响急性血管损伤后的形成和重新皮层化。我们最近确定了VSMC特异性抗迁移的媒介程序可以防止VSMC迁移而不会影响EC迁移。我们确定通过对PDGFR-β激活进行VSMC抗迁移式APATMER操作，但对PDGF-BB-没有影响介导的VSMC增殖。这些数据表明了PDGFR-β迁移和增殖的新概念信号传导可能会解离。现在，我们已经确定了调节的另外两个VSMC靶向适体 VSMC，但不是EC，通过源自细胞表面的未知机制，增殖和凋亡。我们将测试总体假设，即VSMC细胞表面蛋白和信号通路所必需的急性血管损伤后的迁移，增殖和凋亡可能会被特异性调节至防止新内膜形成，而不会改变EC重新皮层化。在AIM 1中，我们将定义机制通过哪些VSMC PDGFR-β依赖性迁移与PDGFR-β依赖性增殖分离，并且可能被抑制以防止新的形成，而不会干扰重新皮化。 AIM 2将确定细胞表面蛋白，使用全基因组CRISPR库的新应用，并定义机制在VSMC中可以特别抑制VSMC增殖以防止新的形成而没有影响重新皮化。 AIM 3将确定VSMC的机制和细胞表面蛋白质凋亡可能被诱导限制新内膜形成，同时保存重新皮层化。完成这项研究将导致对调节VSMC迁移，增殖和凋亡的机械理解要防止新内膜形成并保存重新皮层化，并适用于许多急性血管疾病。