Biomimetic Vascular Matrix for Vascular Smooth Muscle Cell Mechanobiology and Pathology

用于血管平滑肌细胞力学生物学和病理学的仿生血管基质

基本信息

批准号：
10683796
负责人：
Yongho Bae
金额：
$ 63.95万
依托单位：
STATE UNIVERSITY OF NEW YORK AT BUFFALO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-09 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10683796
关键词：
3-Dimensional Address Affect Amino Acids Animal Model Aorta Apoptosis Inhibitor Arterial Injury Arteries Atherosclerosis Atomic Force Microscopy Attenuated Biochemical Biocompatible Materials Biological Biological Models Biology Biomechanics Biomimetics Biophysical Process Blood Vessels Cardiovascular Diseases Cardiovascular system Cell physiology Cells Cellular biology Chromatin Collagen Coronary Arteriosclerosis Coronary heart disease Coupled DNA Sequence Alteration Data Development Disease Disease Progression Engineering Environment Event Extracellular Matrix Extracellular Matrix Proteins Family suidae Feedback Fibronectins Fluorescent in Situ Hybridization Gene Expression Genetic Transcription Goals Histologic Human Hyperplasia In Vitro Injury Knock-out Label Machine Learning Mechanics Mediating Medicine Microscopy Modeling Molecular Monitor Morphology Mus Nuclear Structure Optics Pathologic Pathology Pharmacotherapy Phenotype Physical condensation Physiological Production Property Proteins RNA Research Research Personnel Research Proposals Resolution Role Smooth Muscle Myocytes Structure System Testing Therapeutic Time Study Tissue Engineering Tissues Up-Regulation Vascular Proliferation Vascular Smooth Muscle Vascular System Work arterial remodeling arterial stiffness base cardiovascular risk factor cell behavior cell motility femoral artery in vivo in vivo Model injured knock-down mRNA Expression machine learning algorithm member microscopic imaging migration mouse model nanofiber nanoscale neointima formation new therapeutic target novel overexpression polyacrylamide hydrogels protein expression reconstruction response scaffold single cell analysis soft tissue survivin targeted treatment therapeutic target three dimensional cell culture transcriptome sequencing vascular abnormality vascular injury vascular smooth muscle cell migration vascular smooth muscle cell proliferation

项目摘要

SUMMARY Arterial stiffness is a key risk factor for cardiovascular disease (CVD) events. Change in arterial stiffness is a significant pathology in vascular injury, atherosclerosis, and coronary disease by which stiffening of the vessel wall promotes anomalous migration and proliferation of vascular smooth muscle cells (VSMCs) causing neointima formation of the vessel wall. Yet, the molecular mechanisms by which pathological ECM stiffness regulates VSMC proliferation and migration associated with pathological ne- ointima formation are unclear. This research proposal will address this gap by exploring how changes in arterial stiffness elicit VSMC behaviors that contribute to CVD. More specifically, this work draws upon newly collected preliminary data that show a novel role for the protein survivin as a key regulator of stiffness-mediated VSMC proliferation and migration and an effector of arterial stiffening and remodel- ing. Using mouse and human VSMCs, this study will first explore how vascular ECM stiffness impacts VSMC migration, proliferation, and chromatin organization at the single-cell level (early stage of disease progression; Aim 1); and, secondly, determine how pathological ECM stiffness drives neointima for- mation altering the local mechanical environment of VSMCs in vitro (advanced stage of disease pro- gression; Aim 2). Lastly, this research proposal will test survivin’s role in regulating both ECM production and arterial stiffness (in vivo animal model; Aim 2). These aims will be achieved using a 3D cell culture using a novel in vitro porcine decellularized aorta ECM based (daECM) fibrous scaffold system and engineered mouse injury models. Briefly, VSMCs isolated from mouse and human aortas will be cultured on daECM-based nanofibrous scaffolds of different stiffnesses that mimic normal and pathological con- ditions in the body. The VSMC responses to pathological ECM stiffness will be analyzed using advanced microscopy to observe changes in cellular/nuclear structure, biomechanical properties, and the RNA and protein expressions at the single-cell level in vitro. Finally, engineered mice will be used to study stiffness and VSMC function in intact arteries, performing a histological examination and biochemical analyses of dissected tissue after stiffness is manipulated by arterial injury, drug treatment, or genetic mutations. This project will, for the first time, study the molecular and biophysical mechanisms by which survivin 1) mediates stiffness-sensitive VSMC functions, and 2) contributes to neointima formation and stiffening, revealing a completely new aspect of survivin biology in VSMCs and in the pathology of arte- rial stiffness. Overall, this proposal is unique in its ability to identify potential new therapeutic targets for the treatment of CVDs.

概括动脉僵硬度是心血管疾病 (CVD) 事件的关键危险因素。是血管损伤、动脉粥样硬化和冠状动脉疾病的重要病理学，通过它使血管硬化血管壁促进血管平滑肌细胞的异常迁移和增殖（VSMC）引起血管壁新内膜形成的分子机制。病理性 ECM 硬度调节与病理性神经相关的 VSMC 增殖和迁移内膜的形成尚不清楚，本研究提案将通过探索如何变化来解决这一差距。更具体地说，这项工作借鉴了动脉硬化引起的 VSMC 行为，从而导致 CVD。新收集的初步数据显示蛋白质生存素作为关键调节因子的新作用硬度介导的 VSMC 增殖和迁移以及动脉硬化和重塑的效应器 - 这项研究将首先利用小鼠和人类 VSMC 探讨血管 ECM 硬度如何影响单细胞水平上的 VSMC 迁移、增殖和染色质组织（疾病早期）目标 1)；其次，确定病理性 ECM 僵硬如何驱动新内膜- 体外改变 VSMC 的局部机械环境（疾病的晚期阶段）回归；最后，本研究计划将测试 Survivin 在调节 ECM 产生中的作用。和动脉僵硬度（体内动物模型；目标 2）将通过 3D 细胞培养来实现。使用新型体外猪脱细胞主动脉 ECM (daECM) 纤维支架系统简而言之，将从小鼠和人类主动脉分离的 VSMC 进行培养。基于 daECM 的具有不同硬度的纳米纤维支架，模拟正常和病理状态将使用先进的技术来分析 VSMC 对病理性 ECM 僵硬的反应。显微镜观察细胞/核结构、生物力学特性和 RNA 的变化最后，将使用工程小鼠进行研究。完整动脉的硬度和 VSMC 功能，进行组织学检查和生化检查通过动脉损伤、药物治疗或遗传控制硬度后对解剖组织进行分析该项目将首次研究突变的分子和生物物理机制。 survivin 1) 介导硬度敏感的 VSMC 功能，2) 有助于新内膜形成和变硬，揭示了 VSMC 和动脉病理学中存活蛋白生物学的全新方面总体而言，该提案的独特之处在于它能够确定潜在的新治疗靶点。 CVD 的治疗。