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: 8913536
• 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/8913536
• 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; Genes; Growth Factor; Heart; Heparin; Human; Hyperplasia; In Vitro; Inflammation; Inflammatory; Insulin-Like Growth Factor I; 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; Pathology; Pathway interactions; Patients; Phosphoric Monoester Hydrolases; Positioning Attribute; Pump; Radiation; Recording of previous events; Research; Resolution; Saphenous Vein; Side; Smooth Muscle Myocytes; Solutions; Stenosis; System; TIMP3 gene; Techniques; Tenascin; Testing; Therapeutic; Thick; Thrombosis; Tissue Inhibitor of Metalloproteinase-3; Touch sensation; Transgenes; Translating; Treatment Efficacy; Ulcer; Veins; Wit; Work; adverse outcome; ascending aorta; cell growth; cytokine; gene therapy; graft failure; heart function; high risk; human disease; injured; intercellular communication; 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（由申请人提供）：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。根本机制仍不清楚。各种疗法（包括靶向基因疗法）的失败可能是由于基因选择不当和/或使用与人类疾病不同的动物模型。另外，也没有考虑促进内皮细胞增殖。我们观察到 PTEN 失活会导致人 SV 中 SMC 增殖增加，并且 PTEN 过度表达会限制细胞增殖。由于 PTEN 调节细胞信号传导和细胞生长，因此 SV 移植 SMC 中 PTEN 转基因表达将阻止内膜增生的发展。此外，用间充质干细胞（MSC）修复内皮细胞（EC）会抑制血栓形成。假设在 CABG 之前通过 PTEN 转基因抑制内膜增生，同时通过在自体静脉移植物中输送 MSC 来保护内皮，将是维持静脉移植物通畅的最佳策略。我们将在我们实验室常规使用的成熟猪动脉粥样硬化模型中进行这些研究，并使用通过“非接触技术”分离的腹壁浅静脉 (SEV) 进行 CABG。目标 1：我们的假设预测，动脉粥样硬化猪冠状动脉静脉移植的 SMC 中 PTEN 转基因的过度表达将阻止旁路静脉移植中新生内膜增生的发生。目标2：我们的假设预测，在动脉粥样硬化猪冠状动脉静脉移植的SMC中，用MSC与PTEN转基因一起修复内皮细胞将预防旁路静脉移植中的血栓形成和新生内膜增生，并且这将优于单独PTEN转基因的效果。目标 3：我们的假设预测，PTEN 转基因的过度表达和 MSC 诱导的内皮细胞再生将减少炎症并抑制收缩性重塑，同时抑制冠状动脉静脉移植物中的新内膜增生。自体 SEV 将暴露于 PTEN 载体和 MSC，然后在动脉粥样硬化猪中进行主动脉冠状动脉移植。用于监测心脏功能的心电图和超声心动图，以及用于量化节段内最小管腔直径、直径狭窄、晚期丢失和内膜增生的冠状动脉造影和最佳相干断层扫描将检查该疗法的治疗效果。组织学参数包括内膜厚度、管腔面积、内膜-中膜比率、斑块的形成和溃疡、血栓形成、细胞外基质和再闭塞。将在切除的静脉移植物中检查促炎和抗炎以及促增生途径的表达，并将其与新内膜增生和收缩重塑的程度相关。 这些研究的结果将为我们的假设提供概念支持，并使我们能够将我们的研究转化为临床 1 期研究，使用 PTEN 工程化的自体 MSC 来治疗和可能治愈 CABG 后的静脉移植疾病。