MESENCHYMAL STEM CELLS IN THE PREVENTION OF THROMBOSIS AND NEOINTIMAL HYPERPLASIA

间充质干细胞预防血栓形成和新生内膜增生

• 批准号: 8257841
• 负责人: Devendra K. Agrawal
• 金额: $ 63.68万
• 依托单位: CREIGHTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-12-01 至 2016-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8257841
• 关键词: Angiography; Angioplasty; Apoptosis; Area; Atherosclerosis; Autologous; Balloon Angioplasty; Blood; Blood Platelets; Blood Vessels; Caliber; Cardiology; Cardiovascular system; Cell Differentiation process; Cell Proliferation; Cell Therapy; Cellular biology; Clinical; Coronary; Coronary Angiography; Coronary Arteriosclerosis; Coronary artery; Coronary heart disease; Data; Deposition; Development; Down-Regulation; Drug Controls; Endothelial Cells; Engineering; Family suidae; Fibrinogen; Functional disorder; Gene Expression; Gene Transduction Agent; Genes; Goals; Growth; Heart; Histology; Human; Hyperlipidemia; Hyperplasia; In Vitro; Inflammatory; Injury; Insulin-Like Growth Factor I; Interferons; Interleukin-12; Investigation; Laboratories; Legal patent; Lipids; Mediator of activation protein; Mesenchymal Stem Cells; Metals; Mitogens; Modeling; Molecular Biology; Optics; Outcome; PTEN gene; Pathology; Patients; Percutaneous Transluminal Coronary Angioplasty; Pharmaceutical Preparations; Phosphoric Monoester Hydrolases; Positioning Attribute; Prevention; Procedures; Receptor Cell; Research; Resolution; Role; Signal Transduction; Signaling Molecule; Site; Smooth Muscle Myocytes; Stem cells; Stenosis; Stents; Symptoms; TNF gene; Testing; Therapeutic; Thick; Thrombosis; Transduction Gene; Transgenes; Translating; Ulcer; Vascular Diseases; Vascular Endothelial Growth Factors; cell growth; cytokine; gene therapy; in vivo; intercellular communication; intima media; novel; overexpression; phase 1 study; prevent; receptor expression; repaired; response to injury; restenosis; shear stress; stem cell differentiation; tomography; transgene expression; vector; vector control

DESCRIPTION (provided by applicant): Drug-eluting stents cause less intimal hyperplasia and less late luminal loss, but inhibit re- endothelialization of the stented segment making it more susceptible to thrombosis. Thus, there is trade-off between restenosis and late stent thrombosis requiring longer periods of anti-platelet therapy. Clearly, a better therapeutic approach that can prevent intimal hyperplasia and induce re-endothelialization would be useful to provide long-term symptom-free control of coronary artery disease without the need for prolonged anti- platelet therapy. Since PTEN modulates cell signaling and cell growth, PTEN transgene overexpression in arterial smooth muscle cells would prevent the development of neointimal hyperplasia. In addition, thrombosis could be prevented by the repair of endothelial cells with mesenchymal stem cells (MSCs) which have been primed to differentiate into endothelial cells. We hypothesize that the delivery of MSCs and PTEN transgene locally to the site of angioplasty and stent injury prevents thrombosis and the development of neointimal hyperplasia. Down-regulation of Wnt/2 catenin signaling in MSCs promotes endothelial cell differentiation and re-endothelialization. This hypothesis has been supported by our data demonstrating that vascular endothelial growth factor (VGEF)-primed MSCs differentiate into endothelial cells and PTEN overexpression inhibits proliferation of arterial smooth muscle cells. We will do these studies in a well-established and routinely used in our laboratory the swine model of hyperlipidemia and atherosclerosis with angioplasty and in-stent restenosis. Aim 1: Test the hypothesis that overexpression of PTEN transgene with MSCs at the site of balloon angioplasty in atherosclerotic coronary artery prevents thrombosis and the development of neointimal hyperplasia. Aim 2: Test the hypothesis that overexpression of PTEN transgene with MSCs at the site of bare metal stenting in atherosclerotic coronary artery prevents thrombosis and in-stent restenosis and this would be superior to the effect of drug-eluting stent alone. Aim 3: Test the hypothesis that down-regulation of Wnt/2-catenin signaling promotes MSC differentiation to endothelial cells at the injury site in coronary arteries. We will examine the role of Wnt/¿-catenin signaling molecules, particularly ¿-catenin, LRP5 and Kremen1, as the underlying mechanism of MSCs differentiation into ECs and re-endothelialization both in vitro and in vivo in swine coronary arteries. These studies would position us to translate our investigation into a clinical phase 1 study for the use of PTEN-engineered autologous MSCs for the treatment of the complications due to coronary interventional procedures, and PTCA with PTEN-engineered autologous MSCs might eliminate the need of stents in coronary artery disease. PUBLIC HEALTH RELEVANCE: Re-narrowing of coronary arteries in the heart after balloon angioplasty or placement of stents is serious problem. This is primarily due to uncontrolled growth of smooth muscle cells at the site of injury due to balloon angioplasty or the placement of stent in coronary artery. Another major problem is the deposition of platelets that develop thrombosis. In this project we proposed to deliver autologous mesenchymal stem cells together with a novel gene at the site of injury in the heart of atherosclerotic pig during these procedures to prevent the re-narrowing of the coronary arteries and keep them patent for longer period. The goal is to test the hypothesis that the administration of stem cells with gene therapy is superior to drug-eluting stents. The proposed studies will provide conceptual support of our hypothesis and position us to translate our investigation into a clinical phase 1 study for the delivery of stem cells with the novel gene in patients with coronary artery disease.

描述（由申请人提供）：药物洗脱支架引起的内膜增生和晚期管腔丢失较少，但抑制了支架段的再内皮化，使其更容易发生血栓形成。因此，在再狭窄和晚期支架血栓形成之间存在权衡，需要更长时间的抗血小板治疗。显然，一种可以预防内膜增生并诱导再内皮化的更好治疗方法将有助于提供冠状动脉疾病的长期无症状控制，而无需长期抗血小板治疗。由于PTEN调节细胞信号传导和细胞生长，因此动脉平滑肌细胞中PTEN转基因过表达将阻止新生内膜增生的发展。此外，可以通过用间充质干细胞（MSC）修复内皮细胞来防止血栓形成，所述间充质干细胞已被诱导分化为内皮细胞。我们假设将MSC和PTEN转基因局部递送到血管成形术和支架损伤部位可以预防血栓形成和新生内膜增生的发生。下调Wnt/2 catenin信号通路促进内皮细胞分化和再内皮化这一假设得到了我们的数据的支持，我们的数据表明，血管内皮生长因子（VGEF）引发的MSC分化为内皮细胞和PTEN过表达抑制动脉平滑肌细胞的增殖。我们将在我们实验室中建立并常规使用的高脂血症和动脉粥样硬化伴血管成形术和支架内再狭窄的猪模型中进行这些研究。目标1：验证在动脉粥样硬化冠状动脉球囊成形术部位用MSC过表达PTEN转基因可预防血栓形成和新生内膜增生的发展的假设。目标二：验证在动脉粥样硬化冠状动脉裸金属支架植入部位用MSC过表达PTEN转基因预防血栓形成和支架内再狭窄的假设，这将上级单独的药物洗脱支架的效果。目标三：检验Wnt/2-catenin信号转导下调促进冠状动脉损伤部位MSC分化为内皮细胞的假设。我们将研究Wnt/<$-连环蛋白信号分子，特别是<$-连环蛋白，LRP 5和Kremen 1的作用，作为MSC分化为EC和再内皮化的潜在机制，在体外和体内猪冠状动脉。这些研究将使我们能够将我们的研究转化为临床1期研究，使用PTEN工程化自体MSC治疗冠状动脉介入手术引起的并发症，并且使用PTEN工程化自体MSC的PTCA可能消除冠状动脉疾病中支架的需要。 公共卫生相关性：球囊血管成形术或支架置入后心脏冠状动脉再狭窄是一个严重的问题。这主要是由于球囊血管成形术或冠状动脉内支架置入导致的损伤部位平滑肌细胞的不受控制的生长。另一个主要问题是血小板的沉积，形成血栓。在这个项目中，我们提出了在这些程序中将自体间充质干细胞与一种新基因一起递送到动脉粥样硬化猪心脏损伤部位，以防止冠状动脉再狭窄并使其保持更长时间的通畅。目的是检验基因治疗的干细胞管理上级药物洗脱支架的假设。拟议的研究将为我们的假设提供概念上的支持，并使我们能够将我们的研究转化为临床1期研究，用于在冠状动脉疾病患者中提供具有新基因的干细胞。