Accelerating Wound Healing through RNAi

描述（由申请人提供）：老年人和II型糖尿病患者人数不断增加，慢性伤口愈合缓慢的风险很高。促进伤口愈合的有效疗法可以显着改善这些人的生活。正常的伤口愈合涉及一系列协调的事件，这些事件部分地由伤口部位的脉管系统损伤引起的缺氧刺激。这些事件包括血管生成、血管发生、巨噬细胞募集、细胞凋亡抑制以及成纤维细胞和角质形成细胞的扩增和动员以用于再上皮化。在慢性伤口中，对缺氧的正常反应受损，许多这些细胞过程受阻。在这个项目中，我们提出了使用RNA干扰（RNAi）和反义方法的组合来调节涉及这些过程的三个关键靶标。在第一阶段，我们确定了寡核苷酸，降低这些目标中的两个（蛋白质和microRNA [miRNA]）的水平，增加促进血管生成的下游因子的水平，并增加角质形成细胞的流动性。其中一种抑制剂使用Somagenics专有的"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，我们将测试第三个目标，这也是一个miRNA的调节水平的影响，并评估这些寡核苷酸的生物学效应时，单独使用或组合在组织培养模型相关的伤口愈合。然后，我们将测试这三种寡核苷酸在小鼠中在正常以及受损（缺血性和糖尿病）伤口愈合条件下的体内功效。为了增强体内稳定性并消除任何不期望的免疫刺激，所有三种寡核苷酸将被化学修饰。此外，还提供了几种不同的方法， 将评价RNA对组织加速伤口闭合的功效和对寡核苷酸靶标以及伤口愈合中涉及的下游因素的影响。这种新的，三管齐下的方法来调节伤口愈合中涉及的因素，可以代表治疗慢性伤口的有效策略，也可能是急性伤口。

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.