GENE THERAPY OF CORONARY ARTERY DISEASE

我们的研究旨在评估基因治疗是否与 复制缺陷的重组腺病毒载体可用于 诱导血管生成，恢复缺血组织的血液供应，并 预防和治疗血管成形术后再狭窄。为了研究关于 血管生成，我们构建了携带cDNA腺病毒载体 针对不同的血管生成生长因子：VEGF165、aFGF1-154、重组 通过添加信号肽修饰分泌的aFGF1-154 分泌成纤维细胞生长因子-4、碱性成纤维细胞生长因子、PD-ECGF。携带cDNA的载体 血管内皮细胞生长因子和碱性成纤维细胞生长因子诱导内皮细胞增殖和 共注射时的体外分化和体内血管生成 与重组基底膜蛋白(Matrigel)结合 在老鼠身上。此外，初步结果显示，自体内皮细胞 AdCMV.VEGF165体外感染细胞，随后注射入 兔肢体缺血模型中的髂动脉供血 后肢缺血在体内诱导血管生成。该项目的建设 其余的向量最近才完成，向量是 在体外对其生物效应进行评估。为了研究关于 血管成形术后的再狭窄我们已经构建了腺病毒载体 可通过以下步骤之一起作用：(1)选择性地增强 血管损伤处内皮细胞再生。(AdCMV.VEGF165 和AdCMV.PDECGF)。(2)诱导血管平滑肌细胞死亡或生长 (AdCMV.WAF-1、AdCMV.IGF1R-AS和AdCMV.MKP-1)。此外，还有一个 携带人野生型P53基因的腺病毒载体已成功构建 从约翰·霍普金斯大学的Vogelstein博士那里获得。(3)抑制 血管平滑肌细胞从中膜向内膜迁移 (AdCMV.TIMP-2)。上述向量目前正在评估中 体外测定其生物学效应。初步结果显示， AdCMV.P53抑制血管平滑肌细胞增殖 而AdCMV.TIMP-2则抑制VSMC在Boyden小室的侵袭。

Our studies are aimed at evaluating whether gene therapy with replication-deficient, recombinant adenovirus vectors can be used to induce angiogenesis and restore blood supply to ischemic tissues and to prevent and treat restenosis after angioplasty. For the studies on angiogenesis, we have constructed adenoviral vectors which carry the cDNA for different angiogenic growth factors: VEGF165, aFGF1-154, recombinant secreted aFGF1-154 modified by the addition of the signal peptide for secretion from FGF-4, bFGF, PD-ECGF. The vectors which carry the cDNAs for VEGF and for aFGF induce endothelial cell proliferation and differentiation in vitro as well as angiogenesis in vivo when coinjected sucutaneously with reconstituted basement membrane proteins (Matrigel) in mice. Further, preliminary results show that autologous endothelial cells infected ex vivo with AdCMV.VEGF165 and subsequently injected into the iliac artery supplying the ischemic limb in a rabbit model of hindlimb ischemia induce angiogenesis in vivo. The construction of the remaining vectors has been completed only recently and the vectors are being evaluated for their biologic effects in vitro. For the studies on restenosis after angioplasty we have constructed adenoviral vectors which may act through one of the following steps: (1) selectively enhance endothelial cell regrowth at the site of vascular injury. (AdCMV.VEGF165 and AdCMV.PDECGF).(2) induce vascular smooth muscle cell death or growth arrest.(AdCMV.WAF-1, AdCMV.IGF1R-AS and AdCMV.MKP-1). In addition an adenovirus vector which carries the cDNA for human wild type p53 has been obtained from Dr. Vogelstein (Johns Hopkins University). (3) inhibit vascular smooth muscle cell migration from the media to the intima (AdCMV.TIMP-2). The above vectors are presently being evaluated to determine their biologic effects in vitro. Preliminary results show that AdCMV.p53 inhibits vascular smooth muscle cells (VSMC) proliferation while AdCMV.TIMP-2 inhibits VSMC invasion in the Boyden chamber.