GENE THERAPY WITH SOCS-3 IN INTIMAL HYPERPLASIA AND IN-STENT RESTENOSIS

描述（由申请人提供）：介入手术后的内膜增生和再狭窄仍然是重要的临床问题。药物洗脱支架导致较少的内膜增生和较少的晚期管腔损失，但抑制支架段的再内皮化，使其更容易形成血栓，需要更长时间的抗血小板治疗。显然，一种额外的治疗方法将有助于预防内膜增生和提供长期无症状的冠状动脉疾病控制。SOCS3调节VSMCs中有丝分裂原的活性，在有丝分裂原和炎症细胞因子同时存在的情况下，SOCS3基因被沉默，导致生长因子的增殖作用增加。这证明在冠状动脉中引入SOCS3基因是控制内膜增生的可行方法。假设将SOCS3转基因局部传递到血管成形术和支架损伤部位，可以防止新生内膜增生和支架内再狭窄的发生，这是由于冠状动脉平滑肌细胞中SOCS3基因的高甲基化沉默和STAT3和NF-kB转录活性的增强。我们建议采用血管成形术和支架内再狭窄的高脂血症和动脉粥样硬化微型猪模型。目的1：我们的假设预测，在动脉粥样硬化的冠状动脉球囊血管成形术部位引入SOCS3转基因将阻止内膜增生的发展。目的2：我们的假设预测，在动脉粥样硬化的冠状动脉裸金属支架植入术部位引入SOCS3转基因可以预防支架内再狭窄，这将优于单独使用药物洗脱支架的效果。在这两个目标中，aav9介导的基因转移将用于在高脂血症和动脉粥样硬化微猪冠状动脉介入手术部位的VSMCs中过表达SOCS3。将检查冠状动脉内膜增生和支架内再狭窄的临床和组织学参数。目的3：我们的假设预测球囊血管成形术和血管内支架植入术后的内膜增生是由于冠状动脉平滑肌细胞中STAT3和NF-kB转录活性的高甲基化沉默了SOCS3启动子，并增强了STAT3和NF-kB的转录活性，通过基因治疗恢复SOCS3导致活性NF-kB和STAT3的下调，诱导VSMCs凋亡和生长抑制。我们将确定CpG岛启动子区域的异常高甲基化是否与球囊血管成形术后冠状动脉内膜病变和支架内狭窄的SMCs中SOCS3基因的转录沉默和活性NF-kB和STAT3的上调有关，以及SOCS3基因治疗是否会抑制这种情况。这些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冠状动脉疾病患者中SOCS3基因传递的临床一期研究。

DESCRIPTION (provided by applicant): Intimal hyperplasia and restenosis following interventional procedures remain significant clinical problems. 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 requiring longer periods of anti-platelet therapy. Clearly, an additional therapeutic approach would be useful to prevent intimal hyperplasia and provide long-term symptom-free control of coronary artery disease. SOCS3 regulates the activity of mitogens in VSMCs, and in the presence of both mitogen and inflammatory cytokines, the SOCS3 gene is silenced, leading to increased proliferative effect of growth factors. This justifies the introduction of the SOCS3 gene in coronary arteries as a viable approach to control intimal hyperplasia. The hypothesis is that delivery of the SOCS3 transgene locally to the site of angioplasty and stent injury prevents the development of neointimal hyperplasia and in-stent restenosis which is due to silencing of the SOCS3 gene by hypermethylation and enhanced STAT3 and NF-kB transcriptional activity in coronary artery smooth muscle cells. We propose to utilize hyperlipidemic and atherosclerotic microswine model with angioplasty and in-stent restenosis. Aim 1: Our hypothesis predicts that the introduction of SOCS3 transgene at the site of balloon angioplasty in atherosclerotic coronary artery will prevent the development of intimal hyperplasia. Aim 2: Our hypothesis predicts that the introduction of SOCS3 transgene at the site of bare metal stenting in atherosclerotic coronary artery will prevent in-stent restenosis and this would be superior to the effect of drug- eluting stent alone. In both these aims, AAV9-mediated gene transfer will be used to overexpress SOCS3 in VSMCs at the site of interventional procedure in coronary arteries of hyperlipidemic and atherosclerotic microswine. Clinical and histological parameters of intimal hyperplasia and in-stent restenosis in coronary arteries will be examined. Aim 3: Our hypothesis predicts that the intimal hyperplasia following balloon angioplasty and intravascular stenting is due to silencing of SOCS3 promoter by hypermethylation and enhanced transcriptional activity of STAT3 and NF-kB in coronary artery smooth muscle cells and that restoration of SOCS3 by gene therapy results in down-regulation of active NF-kB and STAT3, induction of apoptosis and growth suppression of VSMCs. We will determine whether or not aberrant hypermethylation of promoter regions in CpG islands is associated with transcriptional silencing of the SOCS3 gene and up- regulation of active NF-kB and STAT3 in SMCs of neointimal lesions in coronary artery following balloon angioplasty and with in-stent stenosis, whether or not this is suppressed by SOCS3 gene therapy. These studies will provide the conceptual support of our hypothesis and position us to translate our investigation into a clinical phase 1 study for the gene delivery of SOCS3 in patients with 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. In this project we proposed to deliver 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 SOCS3 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 gene delivery in patients with coronary artery disease.