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、前景广阔的过程。 用于治疗外周肢体缺血和心肌缺血等疾病 这可能会影响超过50%的并存糖尿病患者。 然而，这些患者中的许多人都被排除在 治疗性血管生成，因为分娩引起的安全问题 和未受调控的血管生成生长因子的表达可能导致 对远端组织有不利影响的溢出效应，例如 视网膜，导致视网膜病变加重。在此应用程序中，我们建议测试 一种新的输送血管生成生长因子的方法，该方法将包含 在目标缺血组织内的物理和临时因素，以及 提供一种更安全的治疗方法，可用于糖尿病和癌症 合并缺血性疾病的患者。 对缺血肢体侧支血管生成的刺激作用及其机制 经血管内皮生长因子和成纤维细胞生长因子蛋白和基因治疗的心肌 已证实发生在动物模型和患者中。肌肉注射 或冠状动脉内注入裸体和腺病毒DNA，已被证明 诱导血管生成，刺激侧支循环，改善组织 功能。该方法治疗脑缺血的治疗潜力 失调症看起来令人鼓舞。在到目前为止的所有研究中，促血管生成基因 都是在强大的病毒启动子的指导下交付的。这些 使用启动子是因为它们很强大，能产生高水平的表达。 主要缺点是：(1)没有规章制度，生产 这些因子由注射剂量、蛋白质的半衰期和 通过率；这排除了更持久的运输工具，如 潜在更有用的腺相关病毒(AAV)。(二)数额大的 注射了质粒DNA或病毒，有很大的溢出风险 对其他促血管生成因子可能有害的组织。溢油已经 已经证实对蛋白质以及质粒和病毒载体都有作用。至 将这些副作用降至最低我们已经开发了组织特异性启动子 受到低氧和消音器元素的强烈调控。我们在这里提议 测试构成启动子可以由这些高度替代的假设 AAV载体中受调控的启动子具有改善的治疗效益和 将副作用的风险降至最低。

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.