GENE THERAPY TO INDUCE THERAPEUTIC ANGIOGENESIS

诱导治疗性血管生成的基因治疗

• 批准号: 2565760
• 负责人: M C CAPOGROSSI
• 金额: --
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2565760
• 关键词: Adenoviridae; angiogenesis; fibroblast growth factor; gene therapy; growth factor; ischemia; laboratory mouse; laboratory rabbit; myocardial infarction; nonhuman therapy evaluation; transfection; vascular endothelial growth factors

The objective of these studies is to determine whether replication-deficient adenovirus vectors which carry the cDNA for angiogenic growth factors may provide a novel approach to induce angiogenesis and enhance collateral blood flow to ischemic tissues in clinically relevant animal models of myocardial and hindlimb ischemia/infarction. We have constructed adenovirus vectors which carry the cDNA for vascular endothelial growth factor (VEGF),for acidic fibroblast growth factor (aFGF) and for a recombinant, secreted form of aFGF. In vitro studies have shown that these vectors enhance endothelial cell proliferation and differentiation into capillary-like structures. In addition, when these vectors were resuspended in reconstituted basement membrane proteins (Matrigel) and the Matrigel was injected subcutaneously in mice, the viral vectors diffused out of the gel and induced angiogenesis in vivo. Subsequently, the therapeutic potential of the vectors coding for VEGF and for secreted aFGF was addressed in rabbit models of hindlimb ischemia and myocardial infarction, respectively. In one study severe hindlimb ischemia was induced by dissecting the femoral artery. Approximately 5 weeks after this surgical procedure autologous endothelial cells were infected ex vivo with the vector coding for VEGF and were injected in the iliac artery supplying the ischemic territory. The cells lodged in the capillaries of the ischemic hindlimb an produced VEGF which led to an improvement in collateral blood flow. In another study the adenovirus vector coding for secreted aFGF was injected along the proximal circumflex coronary artery. Approximately 2-3 weeks later the coronary artery was ligated near the site where the adenovirus vector had been injected and the area at risk for myocardial infarction was assessed with Monastral blue. The group injected with the vector coding for secreted aFGF exhibited a 50% decrease in the area at risk for myocardial infarction vs. both infected and uninfected controls.

这些研究的目的是确定是否 复制缺陷型腺病毒载体，其携带用于 血管生成生长因子可能提供一种新的方法， 血管生成和增强侧支血流到缺血组织， 心肌和后肢的临床相关动物模型 缺血/梗塞。 我们已经构建了携带 血管内皮生长因子（VEGF）cDNA，酸性 成纤维细胞生长因子（aFGF）和重组分泌形式的 aFGF。 体外研究表明，这些载体增强了内皮细胞 细胞增殖和分化成毛细血管样结构。 在 此外，当这些载体重悬于复溶基底中时， 膜蛋白（Matrigel），并将Matrigel注射到 在小鼠皮下，病毒载体扩散出凝胶， 在体内诱导血管生成。 随后， 编码VEGF和分泌型aFGF的载体在 兔后肢缺血和心肌梗死模型。 在一项研究中，严重的后肢缺血是通过解剖 股动脉 手术后大约5周 自体内皮细胞用编码 血管内皮生长因子，并注射在髂动脉供应缺血 领土 这些细胞滞留在缺血后肢的毛细血管中， 产生VEGF，导致侧支血流的改善。 在 另一项研究是编码分泌型aFGF的腺病毒载体， 沿着近端回旋冠状动脉注射。 约2-3 几周后，冠状动脉被结扎的部位附近， 腺病毒载体已经注射，心肌梗死的风险区域 用单星蓝评估梗死。 注射了 编码分泌型aFGF的载体显示，在该区域减少了50% 与感染和未感染的对照相比，心肌梗死的风险。