GENE THERAPY OF CORONARY ARTERY DISEASE

我们的研究旨在评估基因治疗是否 复制缺陷型重组腺病毒载体可用于 诱导血管生成并恢复缺血组织血液供应， 预防和治疗血管成形术后再狭窄。 为研究 血管生成，我们已经构建了携带cDNA的腺病毒载体， 用于不同的血管生成生长因子：VEGF 165、aFGF 1 -154、重组 通过添加信号肽修饰的分泌的aFGF 1 -154， FGF-4、bFGF、PD-ECGF的分泌。 携带cDNA的载体 VEGF和aFGF诱导内皮细胞增殖， 体外分化以及体内血管生成 含重组基底膜蛋白（Matrigel）的支架 对小鼠此外，初步结果表明，自体内皮细胞 用AdCMV. VEGF 165离体感染的细胞，随后注射到 髂动脉供应缺血肢体在兔模型， 后肢缺血诱导体内血管生成。 的建设 其余的矢量最近才完成，矢量是 正在评估其体外生物学效应。 为研究 我们构建了腺病毒载体， 可以通过以下步骤之一起作用：（1）选择性地增强 血管损伤部位的内皮细胞再生。（AdCMV. VEGF 165 和AdCMV.PDECGF）。(2)诱导血管平滑肌细胞死亡或生长 逮捕了（AdCMV.WAF-1、AdCMV.IGF1R-AS和AdCMV.MKP-1）。此外， 携带人野生型p53的cDNA的腺病毒载体已经被 从Vogelstein博士（约翰霍普金斯大学）获得。(3)抑制 血管平滑肌细胞从中膜向内膜迁移 TIMP-2）。 目前正在评估上述载体， 测定其体外生物学效应。 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Boyden小室中VSMC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.