Accelerating Wound Healing through RNAi

描述(申请人提供)：日益增长的老年人口和患有II型糖尿病的人发展成慢性伤口的风险很高，愈合速度很慢。促进伤口愈合的有效疗法可以显著改善这些人的生活。正常的伤口愈合涉及一系列协调的事件，这些事件的部分刺激是由于伤口处血管系统的损伤而导致的缺氧。这些事件包括血管生成，血管生成，巨噬细胞募集，抑制细胞凋亡，以及纤维母细胞和角质形成细胞的扩张和动员，以实现再上皮化。在慢性伤口中，对缺氧的正常反应受到损害，其中许多细胞过程受到阻碍。在这个项目中，我们建议使用RNA干扰(RNAi)和反义方法相结合的方法来调节参与这些过程的三个关键靶点。在第一阶段，我们确定了可以降低其中两个靶点(一种蛋白质和一种微小RNA[miRNA])水平的寡核苷酸，增加促进血管生成的下游因子水平，并增加角质形成细胞的流动性。其中一种抑制剂使用Somagenics的专利sshRNA(短合成发夹RNA)设计，该设计已被证明在治疗感染丙型肝炎病毒的嵌合小鼠方面非常有效。在第二阶段，我们将测试第三个靶点(也是miRNA)的调节水平的效果，并评估这些寡核苷酸单独或联合用于与伤口愈合相关的组织培养模型时的生物学效应。然后，我们将测试这三种寡核苷酸在正常和受损(缺血和糖尿病)小鼠伤口愈合条件下的体内疗效。为了增强体内的稳定性和消除任何不必要的免疫刺激，所有三种寡核苷酸都将进行化学修饰。此外，几种不同的交付方法 将评估组织RNA在加速伤口闭合方面的有效性，以及对寡核苷酸靶标和参与伤口愈合的下游因素的影响。这种新颖的、三管齐下的方法来调节参与伤口愈合的因素，可能代表着一种治疗慢性伤口的有效策略，也可能是治疗急性伤口的有效策略。

DESCRIPTION (provided by applicant): The growing populations of the elderly and those with type-II diabetes are at high risk for developing chronic wounds that are slow to heal. Effective therapeutics to promote wound healing could significantly improve the lives of these people. Normal wound healing involves a coordinated cascade of events that are stimulated in part by the hypoxia that results from injury to the vasculature at the wound site. These events include angiogenesis, vasculogenesis, macrophage recruitment, inhibition of apoptosis, and the expansion and mobilization of fibroblasts and keratinocytes for re- epithelializaton. In chronic wounds, the normal response to hypoxia is impaired and many of these cellular processes are hindered. In this project, we have proposed to modulate three key targets involved these processes using a combination of RNA interference (RNAi) and antisense approaches. In Phase I we identified oligonucleotides that reduce levels of two of these targets (a protein and a microRNA [miRNA]), increase levels of downstream factors promoting angiogenesis, and increase the mobility of keratinocytes. One of these inhibitors uses Somagenics' proprietary sshRNA" (short synthetic hairpin RNA) design, which has been shown to be highly effective in treating chimeric mice infected with hepatitis C virus. In Phase II, we will test the effects of modulating levels of the third target, which is also a miRNA, and evaluate the biological effects of these oligonucleotides when used individually or in combinations in tissue culture models relevant to wound healing. We will then test the in vivo efficacy of these three oligonucleotides under normal as well as compromised (ischemic and diabetic) wound healing conditions in mice. To enhance in vivo stability and eliminate any undesirable immune stimulation, all three oligonucleotides will be chemically modified. In addition, several different methods for delivering RNA to tissue will be evaluated for efficacy in speeding wound closure and effect on the targets of the oligonucleotides as well as downstream factors involved in wound healing. This novel, three-pronged approach to modulating the factors involved in wound healing could represent an effective strategy for treatment of chronic wounds, and potentially also acute wounds.