Therapeutic angiogenesis to treat ischemic disorders

描述（由申请人提供） 治疗性血管生成是一个发展迅速的程序，具有很高的承诺 用于治疗外周肢体缺血和心肌缺血疾病， 这可能会影响超过50%的合并糖尿病的患者。 然而，这些患者中的许多人已被排除在临床试验之外。 治疗性血管生成由于递送导致的安全性问题 和表达不受调节的血管生成因子， 对远端组织产生不利影响的溢出，包括例如 视网膜，导致视网膜病变加重。在本申请中，我们建议测试 一种递送血管生成生长因子的新方法， - 靶缺血组织内的物理和暂时因素，以及 提供一种更安全的治疗方法，可用于糖尿病和癌症 并发缺血性疾病的患者。 刺激缺血肢体侧支血管生成与缺血侧支循环的关系 通过用VEGF和FGF蛋白和基因治疗心肌， 在动物模型和患者中证实发生。肌内注射 或冠状动脉内输注裸DNA和腺病毒DNA，已被证明， 诱导血管生成、刺激侧支循环和改善组织 功能这种方法用于治疗缺血性心脏病的治疗潜力 疾病看起来令人鼓舞。在迄今为止的所有研究中，促血管生成基因 都是在强病毒启动子的指导下进行的。这些 使用启动子是因为它们是强大的并产生高水平表达。 主要缺点是：（1）没有规章制度， 所述因子由注射剂量、蛋白质的半衰期和 清除率;这排除了一个更永久的运载工具，如 可能更有用的腺相关病毒（AAV）。(2)大量 使用质粒DNA或病毒，存在很大的溢出风险 促血管生成因子可能有害的其他组织。溢出效应 已被证明用于蛋白质以及质粒和病毒载体。到 最小化这些副作用，我们开发了组织特异性启动子， 受到缺氧和沉默因子的强烈调节。我们在此提议， 测试组成型启动子可以被这些高度保守的启动子替代的假设。 具有改善的治疗益处的AAV载体中的受调控启动子， 最大限度地减少副作用的风险。

DESCRIPTION (provided by applicant) Therapeutic angiogenesis is a rapidly developing procedure with high promise for the treatment of peripheral limb ischemia and myocardial ischemia, diseases that may affect more than 50 percent of patients with coexisting diabetes. However many of these patients have been excluded from clinical trails of therapeutic angiogenesis because of safety issues resulting from the delivery and expression of unregulated angiogenic growth factors that may result in spillover with adverse effects on distal tissues including for example the retina, causing exacerbated retinopathy. In this application we propose to test a novel method of delivering angiogenic growth factors that will contain the factors physically and temporarily within the target ischemic tissue, and provide a safer treatment that can be used on diabetic as well as cancer patients with complicating ischemic disorders. Stimulation of collateral vessel production in ischemic limbs and the myocardium by treatments with VEGF and FGF proteins and genes has been demonstrated to occur in animal models and patients. Intramuscular injections or intracoronary infusions of naked and adenoviral DNA, have been shown to induce angiogenesis, stimulate collateral circulation, and improve tissue function. The therapeutic potential of this approach for treating ischemic disorders looks encouraging. In all of the studies so far, pro-angiogenic genes have been delivered under the direction of strong viral promoters. These promoters are used because they are powerful and produce high level expression. The principal disadvantages are: (1) There is no regulation, and production of the factors is determined by the injection dose, half life of the protein, and rate of clearance; this precludes a more permanent delivery vehicle such as the potentially more useful adeno associated virus (AAV). (2) Large amounts of plasmid DNA or virus are administered and there is a strong risk of spill-over to other tissues where pro-angiogenic factors may be harmful. Spill-over has been demonstrated for proteins as well as both plasmid and viral vectors. To minimize these side effects we have developed tissue-specific promoters that are strongly regulated by hypoxia and silencer elements. We propose here to test the hypothesis that constitutive promoters can be replaced by these highly regulated promoters in AAV vectors with improved therapeutic benefit and minimized risk of side effects.