GENE AND STEM CELL THERAPY IN CORONARY ARTERY BYPASS GRAFT

冠状动脉搭桥术中的基因和干细胞治疗

• 批准号: 9234420
• 负责人: Devendra K. Agrawal
• 金额: $ 72.58万
• 依托单位: CREIGHTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-03-13 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9234420
• 关键词: Affect; Angiotensin-Converting Enzyme Inhibitors; Animal Model; Animals; Anterior; Anti-Inflammatory Agents; Anti-inflammatory; Aortic coarctation; Area; Arterial Fatty Streak; Arteries; Atherosclerosis; Autologous; Blood Vessels; Bypass; CCL2 gene; Caliber; Cardiology; Cardiovascular system; Cell Proliferation; Cell Therapy; Cellular biology; Cessation of life; Clinical; Clinical Research; Coronary; Coronary Angiography; Coronary Arteriosclerosis; Coronary Artery Bypass; Coronary artery; Development; Disease; Echocardiography; Electrocardiogram; Endothelial Cells; Endothelium; Engineering; Epigastric; Epoprostenol; Event; Extracellular Matrix; Failure; Family suidae; Gene Delivery; Gene Expression; Gene Targeting; Genes; Growth Factor; Heart; Heparin; Histologic; Human; Hyperplasia; In Vitro; Inflammation; Inflammatory; Insulin-Like-Growth Factor I Receptor; Intervention; Investigation; Laboratories; Left; Lipids; Mammary gland; Mediator of activation protein; Mesenchymal Stem Cells; Mitogens; Modeling; Molecular; Molecular Biology; Monitor; Morbidity - disease rate; Myocardial Infarction; Natural regeneration; Operative Surgical Procedures; Outcome; PTEN gene; Pathologic; Pathology; Pathway interactions; Patients; Phosphoric Monoester Hydrolases; Positioning Attribute; Pump; Radiation; Recording of previous events; Research; Resolution; Saphenous Vein; Side; Signal Transduction; Smooth Muscle Myocytes; Stenosis; System; TIMP3 gene; Techniques; Tenascin; Testing; Therapeutic; Thick; Thrombosis; Touch sensation; Transgenes; Translating; Treatment Efficacy; Ulcer; Vein graft; Veins; Wit; adverse outcome; ascending aorta; cell growth; constriction; cytokine; gene therapy; graft failure; heart function; high risk; human disease; injured; internal thoracic artery; intima media; mortality; novel; overexpression; phase 1 study; prevent; public health relevance; receptor expression; repaired; response to injury; restenosis; shear stress; stem cell therapy; tomography; transgene expression; vector; Affect; Angiotensin-Converting Enzyme Inhibitors; Animal Model; Animals; Anterior; Anti-Inflammatory Agents; Anti-inflammatory; Aortic coarctation; Area; Arterial Fatty Streak; Arteries; Atherosclerosis; Autologous; Blood Vessels; Bypass; CCL2 gene; Caliber; Cardiology; Cardiovascular system; Cell Proliferation; Cell Therapy; Cellular biology; Cessation of life; Clinical; Clinical Research; Coronary; Coronary Angiography; Coronary Arteriosclerosis; Coronary Artery Bypass; Coronary artery; Development; Disease; Echocardiography; Electrocardiogram; Endothelial Cells; Endothelium; Engineering; Epigastric; Epoprostenol; Event; Extracellular Matrix; Failure; Family suidae; Gene Delivery; Gene Expression; Gene Targeting; Genes; Growth Factor; Heart; Heparin; Histologic; Human; Hyperplasia; In Vitro; Inflammation; Inflammatory; Insulin-Like-Growth Factor I Receptor; Intervention; Investigation; Laboratories; Left; Lipids; Mammary gland; Mediator of activation protein; Mesenchymal Stem Cells; Mitogens; Modeling; Molecular; Molecular Biology; Monitor; Morbidity - disease rate; Myocardial Infarction; Natural regeneration; Operative Surgical Procedures; Outcome; PTEN gene; Pathologic; Pathology; Pathway interactions; Patients; Phosphoric Monoester Hydrolases; Positioning Attribute; Pump; Radiation; Recording of previous events; Research; Resolution; Saphenous Vein; Side; Signal Transduction; Smooth Muscle Myocytes; Stenosis; System; TIMP3 gene; Techniques; Tenascin; Testing; Therapeutic; Thick; Thrombosis; Touch sensation; Transgenes; Translating; Treatment Efficacy; Ulcer; Vein graft; Veins; Wit; adverse outcome; ascending aorta; cell growth; constriction; cytokine; gene therapy; graft failure; heart function; high risk; human disease; injured; internal thoracic artery; intima media; mortality; novel; overexpression; phase 1 study; prevent; public health relevance; receptor expression; repaired; response to injury; restenosis; shear stress; stem cell therapy; tomography; transgene expression; vector

 DESCRIPTION (provided by applicant): Saphenous vein (SV) graft failure is a common clinical problem in patients undergoing coronary artery bypass graft (CABG) surgery. Underlying mechanisms are still unclear. Failure of various therapies, including targeted gene therapy, could be due to the bad choice of the gene and/or the use of animal model dissimilar to human disease. Also, there was no consideration to promote endothelial cell proliferation. We observed inactivation of PTEN with increased proliferation of SMCs in human SV and PTEN overexpression limits cell proliferation. Since PTEN modulates cell signaling and cell growth, PTEN transgene expression in SV graft SMCs would prevent the development of intimal hyperplasia. Also, the repair of endothelial cells (ECs) with mesenchymal stem cells (MSCs) would inhibit thrombosis. The hypothesis is that the inhibition of intimal hyperplasia by PTEN transgene while preserving endothelium with the delivery of MSCs in autologous vein grafts prior to CABG would be the best strategy to maintain vein graft patency. We will do these studies in a well-established and routinely used in our laboratory the swine model of atherosclerosis and perform CABG using superficial epigastric vein (SEV) isolated with "no touch technique". Aim 1: Our hypothesis predicts that the overexpression of PTEN transgene in the SMCs of vein graft in coronary arteries in atherosclerotic swine will prevent the development of neointimal hyperplasia in bypass vein graft. Aim 2: Our hypothesis predicts that the repair of endothelial cells with MSCs together with PTEN transgene in the SMCs of vein graft in coronary arteries of atherosclerotic swine will prevent thrombosis and neointimal hyperplasia in bypass vein graft, and this would be superior to the effect of PTEN transgene alone. Aim 3: Our hypothesis predicts that overexpression of PTEN transgene and MSCs-induced endothelial cell regeneration would reduce inflammation and inhibit constrictive remodeling together with inhibition of neointimal hyperplasia in the vein graft in coronary arteries. The autologous SEV will be exposed to PTEN vector and MSCs followed by aorto-coronary grafting in atherosclerotic swine. EKG and echocardiography to monitor heart function, and coronary angiography and optimal coherence tomography to quantify in-segment minimal luminal diameter, diameter stenosis, late loss and intimal hyperplasia will examine the therapeutic efficacy of this therapy. Histological parameters will include the intimal thickness, lumen area, intima-media ratio, development and ulceration of plaque, thrombosis, extracellular matrix, and re-occlusion. Expression of pro- and anti-inflammatory and pro-hyperplasia pathways will be examined in excised vein graft and correlated with the degree of neointimal hyperplasia and constrictive remodeling. Findings from these studies will provide the conceptual support for our hypothesis, and position us to translate our investigation into a clinical phase 1 study for the use of PTEN-engineered autologous MSCs for the treatment and possibly cure of the vein graft disease following CABG.

 描述（由适用提供）：大隐静脉（SV）移植失败是接受冠状动脉搭桥术（CABG）手术的患者的常见临床问题。基本机制仍然不清楚。各种疗法的失败，包括靶向基因疗法，可能是由于基因的不良选择和/或使用与人类疾病不同的动物模型的使用。而且，没有考虑促进内皮细胞增殖的考虑。我们观察到PTEN失活，SMC在人类SV中的增殖增加，PTEN过表达限制了细胞的增殖。由于PTEN调节细胞信号传导和细胞生长，因此SV移植SMC中的PTEN转化表达将阻止内膜增生的发展。同样，用间充质干细胞（MSC）修复内皮细胞（EC）将抑制血栓形成。假设是，通过PTEN变换对内膜增生的抑制作用，同时通过在自体静脉图形中保存森林，在自体静脉图形中递送Hothotherium是维持静脉图形的最佳策略。我们将在实验室中建立良好且经常使用的动脉粥样硬化模型中进行这些研究，并使用用“无触摸技术”分离的表面上腹静脉（SEV）进行CABG。目标1：我们的假设预测，即动脉粥样硬化猪中冠状动脉动脉中PTEN转化的过度表达将阻止旁路静脉移植物中新内膜增生的发展。 AIM 2：我们的假设预测，在动脉粥样硬化猪的冠状动脉动脉中，用MSCS的内皮细胞以及PTEN转化以及PTEN转化将防止旁路静脉移植物中的血栓形成和新内膜增生，这将是单独使用PTEN变换的效果。 AIM 3：我们的假设预测，PTEN转基因和MSC诱导的内皮细胞再生的过表达将减少炎症并抑制狭窄的重塑，并抑制冠状动脉中静脉移植物中的新内膜增生。自体SEV将暴露于PTEN载体和MSCS，然后在动脉粥样硬化游泳中进行主动脉冠状术。心脏和超声心动图监测心脏功能，以及冠状动脉造影和最佳相干断层扫描，以量化段内的腔内直径最小，直径，直径狭窄，晚期损失和内膜增生将检查该疗法的治疗效率。组织学参数将包括内膜厚度，管腔面积，内膜媒体比率，牙菌斑，血栓形成，细胞外基质和重新批准的发育和溃疡。促炎和促炎和促炎和促疾病的表达将以优异的静脉移植物进行检查，并与新生素增生和收缩重塑的程度相关。这些研究的发现将为我们的假设提供概念支持，并将我们的投资转化为临床1阶段研究，用于使用PTEN工程的自体MSC，以治疗和治愈CABG后的静脉移植疾病。