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

• 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
• 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

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 动脉僵硬是心血管疾病（CVD）事件的关键危险因素。动脉刚度的变化 是血管损伤，动脉粥样硬化和冠状动脉疾病的重要病理 血管壁促进血管平滑肌细胞的异常迁移和增殖 （VSMC）引起血管壁的新形成。然而，分子机制 病理ECM刚度调节VSMC增殖和与病理NE-相关的迁移 Ointima的形成尚不清楚。该研究建议将通过探索如何改变来解决这一差距 在动脉刚度中，会引起对CVD有助的VSMC行为。更具体地说，这项工作借鉴了 新收集的初步数据显示了蛋白质属蛋白作为作为关键调节剂的新作用 刚度介导的VSMC增殖和迁移以及动脉僵硬和重塑的效应因子 ing。使用鼠标和人类VSMC，该研究将首先探讨血管ECM刚度如何影响 在单细胞水平（疾病的早期阶段）VSMC迁移，增殖和染色质组织 进展；目标1）;其次，确定病理ECM刚度如何驱动新内膜 在体外改变VSMC的局部机械环境（疾病的晚期阶段 草;目标2）。最后，该研究建议将测试Survivin在确定ECM生产中的作用 和动脉刚度（体内动物模型；目标2）。这些目标将使用3D细胞培养 使用一种新型的体外猪脱细胞主动脉ECM（DAECM）纤维支架系统和 工程的小鼠伤害模型。简而言之，将培养从小鼠和人主动脉分离的VSMC 在基于DAECM的纳米纤维支架上，具有模仿正常和病理学的不同刚度 体内的区分。 VSMC对病理ECM刚度的反应将使用高级分析 显微镜观察细胞/核结构，生物力学特性和RNA的变化 在体外单细胞水平的蛋白质表达。最后，工程老鼠将用于学习 完整动脉中的僵硬和VSMC功能，进行组织学检查和生化 通过动脉损伤，药物治疗或遗传来操纵刚度后解剖组织的分析 突变。该项目将首次研究分子和生物物理机制 survivin 1）培养给对刚度敏感的VSMC功能，2） 僵硬，揭示了VSMC和艺术病理中Survivin Biology的全新方面 僵硬。总体而言，该建议在确定潜在的新治疗靶点的能力方面是独一无二的 CVD的处理。