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: 10586599
• 负责人: Yongho Bae
• 金额: $ 58.84万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10586599
• 关键词: 3-Dimensional; Abnormal Cell; Affect; Amino Acids; Animal Model; Aorta; 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 Separation; Cell physiology; Cells; Cellular biology; Chromatin; Coronary Arteriosclerosis; Coronary heart disease; Coupled; DNA Sequence Alteration; Data; Development; Disease; Disease Progression; Environment; Event; Extracellular Matrix; Extracellular Matrix Proteins; Family suidae; Feedback; Fluorescent in Situ Hybridization; Gene Expression; Genetic Transcription; Goals; Histologic; Histology; Human; Hyperplasia; Image; In Vitro; Injury; Invaded; Machine Learning; Mechanics; Mediating; Medicine; Microscopy; Modeling; Molecular; Monitor; Morphology; Mus; Nuclear Structure; Optics; Pathologic; Pathologic Processes; Pathology; Pharmacotherapy; Phenotype; Physical condensation; Physiological; Production; Proliferating; Property; Protein Family; Proteins; RNA; Research; Research Personnel; Research Proposals; Resolution; Role; Smooth Muscle Myocytes; Structure; System; Testing; Therapeutic; Time; Tissue Engineering; Tissues; Vascular Smooth Muscle; Work; arterial remodeling; arterial stiffness; cardiovascular risk factor; cell behavior; cell motility; epigenomics; femoral artery; in vivo; in vivo Model; inhibitor-of-apoptosis protein; injured; knock-down; machine learning algorithm; member; 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; three dimensional cell culture; transcriptome sequencing; transcriptomics; 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. A change in arterial stiffness is a significant pathology in vascular injury, atherosclerosis, and coronary disease. Stiffening of the vessel wall promotes anomalous migration and proliferation of vascular smooth muscle cells (VSMCs), leading to neointima formation on the vessel wall. It is not clear, however, how the extracellular matrix (ECM) influences these pathological processes. This research proposal will address this by exploring how changes in arterial stiffness elicit VSMC behaviors that contribute to cardiovascular disease. Specifically, this work draws upon preliminary data revealing that the protein survivin is a key regulator of stiffness-mediated VSMC proliferation and migration and an effector of arterial stiffening and remodeling. Using mouse and human VSMCs, we 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, second, determine how pathological ECM stiffness drives neointima formation, altering the local mechanical environment of VSMCs in vitro (advanced stage of disease progression; Aim 2). Lastly, we will confirm survivin’s role in regulating both ECM production and arterial stiffness (in vivo animal model; Aim 2). These aims will be achieved using 3D cell culture with a novel in vitro porcine decellularized aorta ECM-based fibrous scaffold system and mouse injury models. Briefly, VSMCs isolated from mouse and human aortas will be cultured on nanofibrous scaffolds of different stiffnesses and structures that mimic normal and pathological conditions in the body. The VSMC responses to pathological ECM stiffness will be analyzed using advanced microscopy to observe changes in cellular/nuclear structure and biomechanical properties in vitro, and the RNA and protein expression will be assessed at the single-cell level. Finally, arterial stiffness and VSMC function will be studied in intact arteries of injured mice; histology and biochemical analyses of dissected tissues will be conducted after arterial stiffness has been 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 (i) mediates stiffness-sensitive VSMC functions and (ii) contributes to neointima formation and stiffening, revealing a completely new aspect of survivin biology in VSMCs and in the pathology of arterial stiffness. Overall, this proposal is unique in its ability to identify potential new therapeutic targets for the treatment of CVDs.

总结 动脉僵硬度是心血管疾病（CVD）事件的关键风险因素。动脉的变化 僵硬是血管损伤、动脉粥样硬化和冠状动脉疾病的重要病理。加强 促进血管平滑肌细胞的异常迁移和增殖 血管平滑肌细胞（VSMCs），导致血管壁新生内膜形成。然而，目前尚不清楚， 细胞外基质（ECM）影响这些病理过程。该研究计划将解决 这是通过探索动脉硬度的变化如何引起VSMC的行为，有助于心血管疾病的发生， 疾病具体来说，这项工作利用了初步数据，揭示了蛋白生存素是一个关键， 刚性介导的VSMC增殖和迁移的调节剂和动脉硬化的效应物 和重塑。使用小鼠和人类VSMCs，我们将首先探讨血管ECM硬度如何影响血管的弹性。 在单细胞水平（早期）影响VSMC迁移、增殖和染色质组织 第二，确定病理性ECM硬度如何驱动新生内膜 形成，改变体外VSMC的局部机械环境（疾病晚期 进展;目标2）。最后，我们将证实生存素在调节ECM产生和动脉粥样硬化中的作用。 刚度（体内动物模型;目标2）。这些目标将使用具有新的细胞培养物的3D细胞培养来实现。 体外猪脱细胞主动脉ECM基纤维支架系统和小鼠损伤模型。简言之， 将从小鼠和人羊膜分离的VSMC培养在不同的纳米纤维支架上。 模拟身体正常和病理条件的硬度和结构。的VSMC 将使用先进的显微镜分析对病理性ECM硬度的反应， 体外细胞/核结构和生物力学特性的变化，以及RNA和蛋白质的变化。 将在单细胞水平评估表达。最后，动脉僵硬度和VSMC功能将 在损伤小鼠的完整动脉中研究;解剖组织的组织学和生物化学分析将在 在动脉硬化已被动脉损伤、药物治疗或遗传操纵后进行 突变。该项目将首次研究分子和生物物理机制， 生存素（i）介导刚度敏感性VSMC功能，（ii）促进新生内膜形成， 硬化，揭示了VSMCs中生存素生物学的一个全新方面， 动脉僵硬总的来说，这个建议是独一无二的，它能够确定潜在的新的治疗靶点 治疗心血管疾病