Accelerating Wound Healing through RNAi

描述（由申请人提供）：老年人口和ii型糖尿病患者的增长是发展慢性伤口的高风险，这些伤口愈合缓慢。促进伤口愈合的有效治疗方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，这些事件部分是由伤口血管损伤引起的缺氧刺激的。这些事件包括血管生成、血管生成、巨噬细胞募集、细胞凋亡抑制、成纤维细胞和角化细胞的扩张和动员以进行再上皮化。在慢性伤口中，对缺氧的正常反应受到损害，许多这些细胞过程受到阻碍。在本项目中，我们建议使用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点。在I期研究中，我们发现寡核苷酸可以降低其中两个靶标（一种蛋白质和一种microRNA [miRNA]）的水平，增加促进血管生成的下游因子的水平，并增加角化细胞的流动性。其中一种抑制剂使用Somagenics专有的sshRNA（短合成发夹RNA）设计，该设计已被证明对治疗感染丙型肝炎病毒的嵌合小鼠非常有效。在第二阶段，我们将测试第三个靶点（也是一个miRNA）的调节水平的影响，并评估这些寡核苷酸在单独使用或在与伤口愈合相关的组织培养模型中联合使用时的生物学效应。然后，我们将测试这三种寡核苷酸在正常和受损（缺血性和糖尿病）小鼠伤口愈合条件下的体内功效。为了提高体内稳定性和消除任何不良的免疫刺激，所有三种寡核苷酸将被化学修饰。此外，还有几种不同的交付方法

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.