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）：使用自体导管进行大约1,000,000个主动脉和周围血管血运重建。移植失败的主要原因是随后的内膜增生。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。该提案将通过防止导致内膜增生的事件来发展一种细胞渗透肽治疗，以增强移植的通畅性。小热休克蛋白HSP27通过涉及p38 MAP激酶和MapKap激酶II（MK2）的激酶级联反应（MK2）磷酸化。磷酸化的HSP27与肌动蛋白应激纤维（肌纤维细胞表型）的形成和增强的平滑肌迁移有关。我们已经开发了一种抑制MK2的细胞渗透肽。该肽还抑制应力纤维形成和ECM产生。该提案的具体目的是：具体目的1：确定可渗透肽的作用，这些肽抑制了HSP27对平滑肌生理，形态和生物化学的磷酸化：我们将确定新型MK2抑制剂肽对完整的人血管平滑肌片段和培养的培养平滑细胞的影响。特定目标＃2：确定优化肽模拟物对内膜增生的影响。我们将首先确定MK2抑制剂对人隐静脉移植器官培养模型中内膜增生模型的影响。随后，我们将确定兔颈动脉介入模型中模拟物在体内的影响。具体目的＃3：确定磷酸化的Hsp27“稳定”肌动蛋白细胞骨架的分子机制：我们将使用定量的高吞吐量质谱技术来分析磷酸化和非磷酸化的HSP27的分子关联。该项目的目的是设计生物分子，该分子使用蛋白质转导域来增强移植物通畅，以将肽直接引入平滑肌细胞中。自体导管代表了这种治疗方法的理想目标，因为可以对移植物进行治疗，从而为交付工程蛋白/肽治疗剂提供了最佳环境。该提案中设计的分子代表了新的治疗剂，因为药物发育的通常靶标（细胞表面受体和信号级联反应）被“绕过”，并且蛋白质蛋白质相互作用通过改变下游靶标蛋白（HSP27）的磷酸化来改变蛋白质蛋白的相互作用。