Local Angiogenic Gene Therapy/VEGF/Treat Diabetic Ulcers

描述（由申请人提供）本提案的目标是确定 如果用ADV-VEGF进行局部血管生成基因治疗可以加速 实验性糖尿病小鼠溃疡。每年，超过90，000个肢体和手指 在美国，84%的糖尿病患者接受截肢手术， 这些都是由一个脚溃疡。与之相关的5年死亡率 截肢率在39%到68%之间。预计年 90亿美元的成本不包括PAM，痛苦和发病率， 糖尿病足溃疡伤口血管生成减少，此外， 神经病变和压力，是延迟愈合的主要致病因素， 这些伤口。目前，没有直接的血管生成疗法可用于 治疗糖尿病足溃疡。VEGF刺激血管生成的能力是很好的， 确立了习由于血管生成在伤口愈合中保持关键作用， 糖尿病伤口中VEGF实验性降低，局部施用VEGF 可以单独或作为组合的一部分提供有效的治疗 治疗糖尿病患者的伤口。局部腺病毒介导基因 治疗提供了持续递送靶点的理论优势 具有最小毒性的分子。尽管VEGF165基因治疗已经被 有效逆转肢体外周缺血、心肌缺血， 糖尿病神经病变，它还没有在神经性糖尿病患者中进行彻底的测试， 溃疡，已知其具有减少的血管生成。我们假设 ADV-VEGF的局部伤口注射将导致VEGF的持续释放， 从而刺激血管生成并加速实验性愈合， 糖尿病小鼠伤口，同时表现出最小的全身副作用。我们 将在6周龄雌性糖尿病小鼠模型中使用ADV-VEGF， 具有可接受的代谢控制，以确定ADV-VEGF的最小剂量 这将导致时间在统计上显著减少到100%， 关闭实验性糖尿病溃疡。我们将衡量潜在的系统性方面 效果（例如，视网膜病变和肝毒性），并确定是否增加 血清中存在VEGF水平。终末器官毒性将通过血清测定 化学、全血细胞计数和肠的组织学分析， 视网膜、肺、肾、脾、胸腺、脑、肝和心脏。我们还将 检查伤口是否有局部毒性PCR将用于评估 腺病毒载体的潜在全身分布。 与变化相比，血管生成反应的特异性定量 将评价胶原沉积和伤口愈合上皮形成。 此外，我们将分析伤口液体以确定时间 VEGF在糖尿病溃疡中的表达。

DESCRIPTION (provided by applicant) The goal of this proposal is to determine if local angiogenic gene therapy with ADV-VEGF accelerates healing in experimental diabetic mouse ulcers. Yearly, over 90,000 limb and digit amputations are performed on diabetic patients in the United States, 84 percent of these are proceeded by a foot ulcer. The 5-year mortality associated with these amputations is between 39 percent and 68 percent. The estimated annual cost of $9 billion doesn't include the pam, suffering, and morbidity of diabetic foot ulcers. Decreased angiogenesis in the wound, in addition to neuropathy and pressure, is a major causative factor for the delayed healing of these wounds. Currently, there is no directly angiogenic therapy available to treat diabetic foot ulcers. VEGF's ability to stimulate angiogenesis is well established. Since angiogenesis maintains a critical role in wound healing and VEGF is experimentally decreased in diabetic wounds, locally administered VEGF may provide an effective treatment, either alone or as part of combination therapy, for patients with diabetic wounds. Local adenoviral mediated gene therapy provides the theoretical advantage of sustained delivery of the target molecule with minimal toxicity. Although VEGF165 gene therapy has been effective in reversing peripheral ischemia of the limbs, cardiac ischemia, and diabetic neuropathy, it has not been thoroughly tested in neuropathic diabetic ulcers, which are known to have decreased angiogenesis. We hypothesize that local wound injection of ADV-VEGF will result in a sustained release of VEGF, thereby stimulating angiogenesis and accelerating healing in experimental diabetic mouse wounds, while demonstrating minimal systemic side effects. We will use ADV-VEGF in the female diabetic mouse model with 6-week mice, which have acceptable metabolic controls, to determine the minimal dose of ADV-VEGF that will result in a statistically significant decrease in time to 100 percent closure m experimental diabetic ulcers. We will measure potential systemic side effects (e.g., retinopathy and liver toxicity) and determine if increased levels of VEGF occur in the serum. End organ toxicity will be assayed by serum chemistries, complete blood count, and histologic analysis of the intestine, retina, lungs, kidney, spleen, thymus, brain, liver and heart. We will also check for signs of local wound toxicity. PCR will be used to assess the potential systemic distribution of the adenoviral vector. Specific quantification of the angiogenic response as compared to the changes in collagen deposition and wound healing epithelialization will be evaluated. In addition, we will analyze the wound fluid in order to determine the temporal expression of VEGF in a diabetic ulcer.