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。药物洗脱支架引起较少的内膜增生和较少的晚期管腔损失，但抑制了支架段的再内皮化，使其更容易发生血栓形成，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防止内膜增生，并提供长期的冠状动脉疾病的无瘤控制。SOCS 3调节VSMC中有丝分裂原的活性，并且在有丝分裂原和炎性细胞因子两者的存在下，SOCS 3基因被沉默，导致生长因子的增殖效应增加。这证明在冠状动脉中引入SOCS 3基因是控制内膜增生的可行方法。该假设是，将SOCS 3转基因局部递送至血管成形术和支架损伤的部位防止了新生内膜增生和支架内再狭窄的发展，这是由于在冠状动脉平滑肌细胞中通过超甲基化和增强的STAT 3和NF-kB转录活性使SOCS 3基因沉默。我们建议利用高血压和动脉粥样硬化的小型猪模型进行血管成形术和支架内再狭窄。目标1：我们的假设预测，在动脉粥样硬化的冠状动脉球囊成形术的位点处引入SOCS 3转基因将防止内膜增生的发展。目标二：我们的假设预测，在动脉粥样硬化冠状动脉裸金属支架植入部位引入SOCS 3转基因将预防支架内再狭窄，这将上级单独的药物洗脱支架的效果。在这两个目标中，将使用AAV 9介导的基因转移在高血压和动脉粥样硬化小型猪的冠状动脉中的介入手术部位的VSMC中过表达SOCS 3。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。目标3：我们的假设是，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SOCS 3启动子的高甲基化和STAT 3和NF-kB在冠状动脉平滑肌细胞中的转录活性增强而引起的，并且通过基因治疗恢复SOCS 3导致活性NF-kB和STAT 3的下调，诱导VSMCs的凋亡和生长抑制。我们将确定CpG岛中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新生内膜病变和支架内狭窄的SMC中SOCS 3基因的转录沉默和活性NF-κ B和STAT 3的上调相关，无论这是否被SOCS 3基因治疗抑制。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SOCS 3基因递送的临床1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.