Prevention of Vein Graft Failure

预防静脉移植失败

基本信息

批准号：
8039215
负责人：
Colleen M Brophy
金额：
$ 42.2万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-04-01 至 2013-12-18
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8039215
关键词：
Actins Arginine Autologous Biochemistry Blood Vessels Bypass Cell Surface Receptors Cells Charge Clinical Cytoskeleton Deposition Development Drug Delivery Systems Engineering Environment Event Extracellular Matrix Goals HSPB1 gene Heat shock proteins Heparin Human Hyperplasia Investigation Lead MAPK14 gene Maps Mass Spectrum Analysis Modeling Molecular Molecular Mechanisms of Action Morphology Muscle function Myofibroblast Organ Culture Techniques Oryctolagus cuniculus Peptides Peripheral Phenotype Phosphorylation Phosphotransferases Physiology Prevention Process Production Prosthesis Protein Engineering Proteins Receptor Signaling Saphenous Vein Smooth Muscle Smooth Muscle Myocytes Stress Fibers Techniques Therapeutic Vascular Smooth Muscle Veins design drug development graft failure in vivo inhibitor/antagonist migration mimetics novel novel therapeutics prevent protein protein interaction response to injury therapeutic target tool

项目摘要

DESCRIPTION (provided by applicant): Approximately 1,000,000 aortocoronary and peripheral vascular revascularizations are performed using autologous conduits. The leading cause of graft failure is the subsequent development of intimal hyperplasia. Intimal hyperplasia represents a response to injury that involves smooth muscle proliferation, migration, phenotypic modulation, and extracellular matrix (ECM) deposition. This proposal will develop a cell permeant peptide therapeutic to enhance graft patency by preventing the events that lead to intimal hyperplasia. A small heat shock protein, HSP27, is phosphorylated by a kinase cascade involving p38 map kinase and MAPKAP kinase II (MK2). Phosphorylated HSP27 is associated with the formation of actin stress fibers (myofibroblast phenotype) and enhanced smooth muscle migration. We have developed a cell permeant peptide that inhibits MK2. This peptide also inhibits stress fiber formation and ECM production. The specific aims of this proposal are: Specific aim #1: Determine the effect of transducible peptides which inhibit the phosphorylation of HSP27 on smooth muscle physiology, morphology, and biochemistry: We will determine the effect of the novel MK2 inhibitor peptide on intact human vascular smooth muscle segments and cultured vascular smooth muscle cells. Specific aim #2: Determine the effect of optimized peptide mimetics on intimal hyperplasia. We will first determine the effect of the MK2 inhibitor on intimal hyperplasia in a human saphenous vein graft organ culture model. Subsequently, we will determine the effect of the mimetics in vivo in a rabbit carotid interposition model. Specific aim #3: Determine the molecular mechanisms by which phosphorylated HSP27 "stabilizes" the actin cytoskeleton: We will use quantitative, high throughput mass spectrometry techniques to analyze the molecular associations of phosphorylated and nonphosphorylated HSP27. The goal of this project is to engineer biomolecules that enhance graft patency using protein transduction domains to directly introduce peptides into smooth muscle cells. Autologous conduits represent an ideal target for this therapeutic approach in that the graft can be treated ex vivo, thus providing an optimal environment for the delivery of engineered protein/peptide therapeutics. The molecules designed in this proposal represent novel therapeutics in that the usual targets of drug development (cell surface receptors and signaling cascades) are "bypassed" and protein-protein interactions are stoichiometrically altered by changing the phosphorylation of downstream target effector proteins (HSP27).

描述（由申请人提供）：大约 1,000,000 例主动脉冠状动脉和外周血管血运重建是使用自体导管进行的。移植失败的主要原因是随后发生的内膜增生。内膜增生代表对损伤的反应，涉及平滑肌增殖、迁移、表型调节和细胞外基质 (ECM) 沉积。该提案将开发一种细胞渗透性肽治疗剂，通过预防导致内膜增生的事件来增强移植物的通畅性。一种小型热休克蛋白 HSP27 被涉及 p38 图谱激酶和 MAPKAP 激酶 II (MK2) 的激酶级联磷酸化。磷酸化 HSP27 与肌动蛋白应激纤维（肌成纤维细胞表型）的形成和增强的平滑肌迁移有关。我们开发了一种抑制 MK2 的细胞渗透肽。该肽还抑制应力纤维的形成和 ECM 的产生。该提案的具体目标是：具体目标#1：确定抑制 HSP27 磷酸化的转导肽对平滑肌生理学、形态学和生物化学的影响：我们将确定新型 MK2 抑制肽对完整人体血管的影响平滑肌片段和培养的血管平滑肌细胞。具体目标#2：确定优化的肽模拟物对内膜增生的影响。我们将首先在人隐静脉移植器官培养模型中确定 MK2 抑制剂对内膜增生的影响。随后，我们将确定模拟物在兔颈动脉介入模型中的体内效果。具体目标#3：确定磷酸化 HSP27“稳定”肌动蛋白细胞骨架的分子机制：我们将使用定量、高通量质谱技术来分析磷酸化和非磷酸化 HSP27 的分子关联。该项目的目标是设计生物分子，利用蛋白质转导域将肽直接引入平滑肌细胞，从而增强移植物的通畅性。自体导管代表了这种治疗方法的理想目标，因为移植物可以在体外进行处理，从而为工程蛋白质/肽治疗剂的递送提供最佳环境。该提案中设计的分子代表了新颖的治疗方法，因为药物开发的常见目标（细胞表面受体和信号级联）被“绕过”，并且通过改变下游靶效应蛋白（HSP27）的磷酸化来化学计量地改变蛋白质-蛋白质相互作用。