Actin and focal adhesion remodeling as therapeutic targets in cardiovascular disease

肌动蛋白和粘着斑重塑作为心血管疾病的治疗靶点

基本信息

批准号：
9303730
负责人：
KATHLEEN G MORGAN
金额：
$ 51.93万
依托单位：
BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-01 至 2022-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9303730
关键词：
Abbreviations Acoustics Actins Acute Adhesions Age Aging Agonist Alzheimer&apos s Disease Amino Acid Sequence Animals Aorta Bioinformatics Biological Assay Biological Markers Biology Biomechanics Blood Pressure Blood Vessels Brain Cardiovascular Diseases Cardiovascular system Cells Chronic Collaborations Colloids Coupled Cytoskeleton Data Databases Defect Dementia Diffusion Magnetic Resonance Imaging Extracellular Matrix Filament Fluorescence Fluorescence Microscopy Focal Adhesions G Actin Goals Heart Heart failure Histology Human Hypertension Hypertrophy Immunoprecipitation Impairment Kidney Kidney Diseases Kidney Failure Lead Lesion Ligation Link Magnetic Resonance Imaging Magnetism Mediating Metabolic Microbubbles Microfilaments Molecular Monitor Mus Muscle Cells Outcome Peptides Phenylephrine Physiologic pulse Protein Isoforms Proteins Publishing Pulse Pressure Recombinant Proteins Recombinants Regulation Research Secondary Hypertension Shock Stress Systole Technology Telemetry Testing Therapeutic Tissues Trauma Ultracentrifugation Ultrasonography Vascular Dementia Vascular Smooth Muscle absorption adverse outcome age effect aged design epidemiology study flexibility hemodynamics in vivo inhibitor/antagonist innovation kidney vascular structure microscopic imaging nanobiotechnology nanoparticle nanoscience novel polymerization prevent programs prototype response small molecule inhibitor targeted delivery therapeutic target vapor vaporization white matter

项目摘要

Recent epidemiological studies have made clear that human proximal aortic stiffness increases with age and is an early and independent biomarker of, and probable contributor to, subsequent adverse cardiovascular outcomes including kidney failure, hypertension and Alzheimer's Disease-related dementia. The normal flexibility of the proximal aorta functions as a critical “shock absorber” to protect small downstream vessels from the high pulses of pressure generated by the heart. We have shown in published studies that the vascular smooth muscle cell (VSMC) regulates up to half of total aortic stiffness and that aging-induced loss of regulation of the VSMC cytoskeleton leads to impairment of the ability of the aorta to perform this shock absorption function. A major advance from our lab has been the demonstration that the cortical nonmuscle actin cytoskeleton and its linkages to focal adhesions and the extracellular matrix are a particularly dynamic and important part of the VSMC cytoskeleton. The broad goal of this program is to test the concept that ultrasound-targeted, cell permeant decoy peptides and small molecule inhibitors can be used both to probe function and to reverse aging-induced malfunction of the VSMC cytoskeleton. We will use cell permeant decoy peptides and recombinant proteins developed by our lab, and for comparison, small molecule inhibitors, to test the hypothesis that ex vivo stiffness of aortas from aged mice can be decreased by mechanisms targeted to the VSMC cytoskeleton. We will mine protein sequence data bases to identify VSMC-specific sequences. Synthetic decoy constructs targeting these sequences will test a proof of concept for the selectivity and efficacy of the peptide approach. We will use biomechanics, magnetic tweezers, proximity ligation analysis, actin polymerization assays and immunoprecipitation to confirm the mechanism of action of decoys. We will test, in collaboration with Tyrone Porter of our Nanoscience Center, the hypothesis that microbubble-packaging of cell-permeant decoys and ultrasound-mediated release will allow localized, tissue-specific targeted delivery of effective decoys. We will test the hypothesis that tissue-targeted decoys can acutely reduce PWV in vivo in mice and that chronic in vivo treatment can prevent 3 negative outcomes associated with aging-induced increased aortic stiffness: brain vascular lesions, hypertension and renal damage. Hence, we propose a highly innovative research strategy to attack aging-induced alterations in the vascular actin cytoskeleton and its linkage to focal adhesions. This approach, no matter the outcome, will answer major questions about aortic stiffness and its relationship to vascular function with age. If successful, this approach has the potential to prevent or reverse a host of aging-associated cardiovascular disorders.

最近的流行病学研究表明，人类近端主动脉僵硬度随着年龄的增长而增加是随后不良心血管事件的早期且独立的生物标志物，并且可能是其促成因素结果包括肾衰竭、高血压和阿尔茨海默病相关的痴呆症。近端主动脉的灵活性作为重要的“减震器”来保护下游的小血管我们在已发表的研究中表明，血管平滑肌细胞 (VSMC) 调节多达一半的总主动脉硬度以及衰老引起的损失 VSMC 细胞骨架的调节失灵会导致主动脉执行此冲击的能力受损我们实验室的一个重大进展是证明皮质非肌肉具有吸收功能。肌动蛋白细胞骨架及其与粘着斑和细胞外基质的联系是一个特别动态的 VSMC 细胞骨架的重要组成部分该计划的总体目标是测试以下概念：超声靶向、细胞渗透性诱饵肽和小分子抑制剂均可用于探测功能并逆转老化引起的 VSMC 细胞骨架功能障碍，我们将使用细胞渗透剂。我们实验室开发的诱饵肽和重组蛋白，为了进行比较，小分子抑制剂，以检验以下假设：老年小鼠主动脉的离体硬度可以通过以下方法降低我们将挖掘蛋白质序列数据库来识别针对 VSMC 细胞骨架的机制。针对这些序列的 VSMC 特异性序列将测试概念验证。为了提高肽方法的选择性和功效，我们将使用生物力学、磁力镊子、邻近连接分析、肌动蛋白聚合测定和免疫沉淀以确认其机制我们将与我们纳米科学中心的蒂龙·波特合作测试这一假设。细胞渗透诱饵的微泡包装和超声介导的释放将允许局部、有效诱饵的组织特异性靶向递送我们将检验组织靶向诱饵的假设。可以急剧降低小鼠体内 PWV，并且长期体内治疗可以预防 3 种负面结果与衰老引起的主动脉僵硬度增加相关：脑血管病变、高血压和肾病因此，我们提出了一种高度创新的研究策略来应对衰老引起的变化。血管肌动蛋白细胞骨架及其与粘着斑的联系，无论结果如何，都会。回答有关主动脉僵硬度及其与年龄的血管功能关系的主要问题。这种方法有可能预防或逆转许多与衰老相关的心血管疾病。