TNF-INHIBITOR GENE THERAPY FOR NEOINTIMAL HYPERPLASIA

肿瘤坏死因子抑制剂基因治疗新生内膜增生

• 批准号: 6537782
• 负责人: KARSTEN PEPPEL
• 金额: $ 38.5万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-06-01 至 2005-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6537782
• 关键词: blood vessel transplantation; cell proliferation; cytokine receptors; dietary lipid; gene therapy; growth inhibitors; hyperplasia; laboratory rabbit; mitogen activated protein kinase; nonhuman therapy evaluation; nutrition related tag; pathogenic diet; restenosis; tumor necrosis factor alpha; vascular endothelium; vascular smooth muscle

Percutaneous coronary intervention (PCI) and coronary artery bypass graft surgery (CABG) are each performed on approximately 500,000 patients annually in the US. Both procedures are complicated by activation, migration and proliferation of cells in the vascular wall, leading to neointimal hyperplasia (NH). A preponderance of evidence suggests that the majority of these cells are of vascular smooth muscle origin. NH affects about 30 % of all PCI patients and necessitates reintervention in some 30 % of all CABG cases within two years. The annual cost needed to treat PCI restenosis alone exceeds $2 billion. PCI and saphenous vein CABG surgery both lead to significant injury of the underlying vessel wall, causing endothelial dysfunction and adherence of platelets, neutrophils and circulating cells of the monocyte / macrophage lineage. Locally released cytokines, such as tumor necrosis factor (TNF), synergize with growth factors, including platelet derived growth factor (PDGF) and basic fibroblast growth factor (bFGF), to activate SMCs in the injured vessel segment. In vitro TNF is a strong mitogen for SMCs and can act in concert with PDGF-BB, to stimulate cell growth. This work proposes to examine the contribution of TNF to the activation of SMCs, alone and in combination with various growth factors. To this end mitogen activated protein kinase (MAPK) activation, DNA synthesis, proliferation, chemotaxis and apoptosis will be examined in TNF treated and untreated SMCs. In addition, this proposal seeks to test the hypothesis that inhibition of TNF can decrease the degree of SMC activation in vitro and can reduce the size of neointima formation in vivo in an experimental animal model. To inhibit TNF, a chimeric TNF receptor extracellular domain - murine immunoglobulin heavy chain construct (TNF-I) was generated and inserted into a recombinant adenovirus (rec. AD-TNF-I). This virus will be used to infect rabbit jugular veins ex vivo that will be grafted across the rabbit's carotid artery in a well characterized model of neointima hyperplasia. Four weeks after bypass grafting the degree of neointima formation in rec.AD-TNF-I and rec.AD-control infected veins will be measured histologically. This will be done both in rabbits receiving a normal diet and also in animals given a high fat diet to examine the contribution of hyperlipidemia towards the development of neointimal hyperplasia in this setting. Reduction of vein graft neointimal hyperplasia by inhibition of TNF may have significant therapeutic implications for human vascular diseases.

在美国，每年约有50万患者接受经皮冠状动脉介入治疗（PCI）和冠状动脉搭桥手术（CABG）。这两个过程都因血管壁细胞的活化、迁移和增殖而复杂化，导致新生内膜增生（NH）。大量证据表明，这些细胞大部分来自血管平滑肌。NH影响约30%的PCI患者，约30%的CABG患者在两年内需要再次干预。每年仅治疗PCI再狭窄所需的费用就超过20亿美元。PCI和隐静脉冠脉搭桥手术都会导致血管壁的严重损伤，导致内皮功能障碍和血小板、中性粒细胞和单核/巨噬细胞循环细胞的粘附。局部释放的细胞因子，如肿瘤坏死因子（TNF），与包括血小板衍生生长因子（PDGF）和碱性成纤维细胞生长因子（bFGF）在内的生长因子协同作用，激活受损血管段的SMCs。在体外，TNF是SMCs的强丝裂原，可以与PDGF-BB协同作用，刺激细胞生长。这项工作旨在研究TNF单独或与各种生长因子联合对SMCs激活的贡献。为此，有丝分裂原活化蛋白激酶（MAPK）的激活、DNA合成、增殖、趋化性和凋亡将在TNF治疗和未治疗的SMCs中进行检测。此外，本提案试图在实验动物模型中验证TNF的抑制可以在体外降低SMC的激活程度，并可以在体内减少新内膜形成的大小。为了抑制肿瘤坏死因子，合成了嵌合肿瘤坏死因子受体细胞外结构域-小鼠免疫球蛋白重链构建体（TNF- i），并将其插入重组腺病毒（AD-TNF-I）中。这种病毒将在体外感染兔颈静脉，并将其移植到兔颈动脉上，形成具有良好特征的新内膜增生模型。旁路移植术4周后，用组织学方法测定recad - tnf - i和recad -control感染静脉的新生内膜形成程度。这将在接受正常饮食的兔子和给予高脂肪饮食的动物中进行，以检查在这种情况下高脂血症对新内膜增生的发展的贡献。通过抑制肿瘤坏死因子减少静脉移植物新生内膜增生可能对人类血管疾病具有重要的治疗意义。