Mechanisms for cell specific modulation of proliferation and migration

细胞特异性调节增殖和迁移的机制

基本信息

批准号：
8413429
负责人：
Thomas S Monahan
金额：
--
依托单位：
BALTIMORE VA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-01-01 至 2016-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8413429
关键词：
Actins Affect Alanine Angioplasty Aortic Injury Area Arterial Injury Baltimore Basic Science Binding Proteins Binding Sites Biological Biological Assay Biology Blood Vessels Bypass Cardiovascular system Cell Adhesion Cell Cycle Cell Proliferation Cells Clinical Communication Complex Coronary Thrombosis Cytoskeleton Cytosol Data Endarterectomy Endothelial Cells Endothelium Future General Population Generations Goals Guanosine Triphosphate Phosphohydrolases Human Hyperplasia In Vitro Incidence Injury Intervention Investigation K-Series Research Career Programs Knock-out Knockout Mice Laboratories Life Ligation Maryland Mediating Medical center Membrane Mentors Modeling Molecular Molecular Genetics Molecular Target Monomeric GTP-Binding Proteins Mus Mutant Strains Mice Null Lymphocytes Operative Surgical Procedures Organ Culture Techniques Pathologic Pathology Pathway interactions Patients Phorbol Esters Phosphorylation Phosphotransferases Plague Price Problem Solving Procedures Protein Kinase Protein Kinase C Proteins Publications Regulation Research Research Ethics Research Personnel Role Saphenous Vein Signal Transduction Site Smooth Muscle Smooth Muscle Myocytes Stents Stress Fibers Surgeon Testing Time Training Transgenic Mice Translational Research United States Department of Veterans Affairs Universities Up-Regulation Work antiproliferative agents antiproliferative drugs base career cell motility cell type clinically relevant design experience in vivo in vivo Model inhibitor/antagonist injured knock-down medical schools migration mouse model mutant operation prevent professor research study response response to injury restenosis skills vascular smooth muscle cell migration vascular smooth muscle cell proliferation

项目摘要

DESCRIPTION (provided by applicant): The proposed research focuses on the formation of intimal hyperplasia, a crucial problem affecting all cardiovascular interventions. Intimal hyperplasia or restenosis results from the pathologic proliferation and migration of smooth muscle cells at the site of vascular injury, caused by surgery or angioplasty. Patients served by the Veterans Administration have a higher incidence of these procedures than the general population. A major limitation of current therapies is that they target not only the vascular smooth muscle cells, but also the beneficial endothelial cells of the injured vessel. The ideal therapy would inhibit vascular smooth muscle cell proliferation and migration without negatively impacting the endothelium. We have identified a promising candidate, the myristolated alanine-rich C kinase substrate (MARCKS), a membrane bound protein that translocates to the cytosol upon phosphorylation by protein kinase C. Previous work by the candidate has demonstrated that MARCKS knockdown results in decreased vascular smooth muscle cell migration and proliferation without affecting endothelial cells. The central hypothesis of this application is that MARCKS represents a cell specific target for modulation of vascular smooth muscle cell proliferation and migration. Currently, the mechanisms by which MARCKS induces vascular smooth muscle cell proliferation and migration are unknown, and the goal of these studies is to elucidate these mechanisms. The specific hypotheses to be tested are: 1) that MARCKS regulates vascular smooth muscle cell proliferation by up- regulation of the cell cycle inhibitor, p27kip1 2) that MACRKS modulates cell migration by disrupting cellular actin organization and 3) that MARCKS knockout in vivo will prevent intimal hyperplasia formation, but allows normal reendothelialization after arterial injury. The proposed studies will elucidate the molecular mechanisms responsible for the differential effects of MARCKS inhibition on smooth muscle cells and endothelial cells noted in the applicant's prior data. These studies will form the scientific basis for smooth muscle cell specific antiproliferative therapy, a significant, unmet need in cardiovascular interventions. The pathways and mechanisms elucidated in the proposed studies will also reveal other potential molecular targets for smooth muscle cell specific antiproliferative and antimigratory therapy. The candidate, Dr. Monahan, is a vascular surgeon at the Baltimore Veterans Administration Medical Center, and an Assistant Professor of Surgery at the University of Maryland School of Medicine. He has previously studied the genetics and molecular mechanisms of intimal hyperplasia, and has numerous publications, including some preliminary data for this proposal. Dr. Monahan's long-term career goal is become an independent clinician-investigator. In order to carry out this research, Dr. Monahan proposes a career development award that is designed to provide him with laboratory skills in several key areas by working with a team of distinguished co-mentors in addition to his mentor, Dr. Wang: the generation and management of complex mouse mutants (Dr. Strickland); the analysis of actin and small GTPase regulation of migration (Dr. Zhan); and the generation of an in vivo model of re-endothelialization (Dr. Sarkar). Dr. Monahan will also receive additional training in laboratory management, research ethics, and scientific communication. These skills are crucial not only to the proposed research, but to Dr Monahan's long term goal: to become an independent clinician-investigator. In summary, the Career Development Award (CDA-2) would provide Dr. Monahan with the research experience, mentoring and skills required to a leader in this important area of translational research. Importantly, Dr. Monahan is fortunate to be part of a larger vascular biology research team, whose support will contribute to his ability to achieve his long-term goal of performing clinically relevant, basic science research.

描述（由申请人提供）：拟议的研究重点是内膜增生的形成，这是影响所有心血管干预措施的一个关键问题。内膜增生或再狭窄是由手术或血管成形术引起的血管损伤部位平滑肌细胞的病理性增殖和迁移引起的。退伍军人管理局服务的患者接受这些手术的发生率高于一般人群。当前疗法的一个主要限制是它们不仅针对血管平滑肌细胞，还针对受损血管的有益内皮细胞。理想的治疗方法是抑制血管平滑肌细胞的增殖和迁移，而不会对内皮细胞产生负面影响。我们已经确定了一个有前途的候选物，即富含肉豆蔻酸化丙氨酸的 C 激酶底物 (MARCKS)，这是一种膜结合蛋白，在被蛋白激酶 C 磷酸化后易位至胞质溶胶。该候选物之前的工作表明，MARCKS 敲低会导致血管平滑度降低肌细胞迁移和增殖而不影响内皮细胞。该申请的中心假设是MARCKS代表调节血管平滑肌细胞增殖和迁移的细胞特异性靶标。目前，MARCKS诱导血管平滑肌细胞增殖和迁移的机制尚不清楚，这些研究的目的是阐明这些机制。要测试的具体假设是：1) MARCKS 通过上调细胞周期抑制剂 p27kip1 调节血管平滑肌细胞增殖 2) MACRKS 通过破坏细胞肌动蛋白组织来调节细胞迁移 3) MARCKS 体内敲除将防止内膜增生形成，但允许动脉损伤后正常的再内皮化。拟议的研究将阐明申请人先前数据中指出的MARCKS抑制对平滑肌细胞和内皮细胞的不同影响的分子机制。这些研究将为平滑肌细胞特异性抗增殖治疗奠定科学基础，这是心血管干预中一个重要的、未满足的需求。拟议研究中阐明的途径和机制还将揭示平滑肌细胞特异性抗增殖和抗迁移治疗的其他潜在分子靶点。候选人莫纳汉博士是巴尔的摩退伍军人管理局医疗中心的血管外科医生，也是马里兰大学医学院的外科助理教授。他之前研究过内膜增生的遗传学和分子机制，并发表了大量出版物，包括该提案的一些初步数据。 Monahan 博士的长期职业目标是成为一名独立的临床医生研究员。为了开展这项研究，莫纳汉博士提出了一项职业发展奖，旨在通过与除他的导师王博士之外的杰出共同导师团队合作，为他提供几个关键领域的实验室技能：复杂小鼠突变体的产生和管理（斯特里克兰博士）；肌动蛋白和小GTP酶迁移调节的分析（詹博士）；以及再内皮化体内模型的生成（Sarkar 博士）。莫纳汉博士还将接受实验室管理、研究伦理和科学传播方面的额外培训。这些技能不仅对拟议的研究至关重要，而且对莫纳汉博士的长期目标至关重要：成为一名独立的临床医生研究员。总之，职业发展奖（CDA-2）将为 Monahan 博士提供成为转化研究这一重要领域的领导者所需的研究经验、指导和技能。重要的是，莫纳汉博士很幸运能够成为更大的血管生物学研究团队的一员，该团队的支持将有助于他实现进行临床相关基础科学研究的长期目标。