MicroRNA mechanism and therapeutics of impaired wound healing

描述（由申请人提供）：伤口愈合受损引起的慢性伤口，特别是糖尿病患者和老年人，仍然是生物医学领域未解决的主要挑战。这突出了研究受损伤口愈合新机制和探索现有治疗方案的重要性和紧迫性。MicroRNAs （miRNAs）是一类具有巨大生物学功能的新型非编码rna。该项目的目的是确定miRNA机制和治疗损伤的伤口愈合。利用基于RCR的阵列和深度测序分析以及qRT-PCR，我们发现了一组在从糖尿病和老年个体和小鼠中分离的骨髓（BM）来源的内皮祖细胞（EPCs）中异常表达的mirna。其中let-7家族成员、let-7b、let-7c、let-7d、let-7d*显著增加。本研究假设Let-7家族成员通过靶基因在糖尿病和老年人EPCs细胞功能受损和伤口愈合受损中发挥重要作用。通过let-7家族调节剂对mirna重编程的“智能”EPCs可能代表了一种新的有效治疗慢性伤口的方法。我们将通过三个具体目标来验证我们的假设：目的1将确定let-7家族成员对体外EPCs迁移、增殖和凋亡的影响。目的2将确定let-7重编程的自体EPCs对糖尿病和老年小鼠体内伤口血管生成和伤口愈合的影响。目的3将确定let-7介导的生物效应的分子机制。let-7家族通过EPCs基因重编程在难治性创伤中的生物学作用是一个尚未探索的研究领域。该研究将利用最新的细胞、分子和全动物方法来解决未解决的主要生物医学挑战：慢性伤口。通过mirna在翻译水平上揭示伤口损伤愈合的新分子机制和治疗方法，可能为慢性伤口研究开辟新的科学范式。

DESCRIPTION (provided by applicant): Chronic wounds induced by the impaired wound healing especially in diabetic patients and aged individuals are still a major unsolved biomedical challenge. This has highlighted the importance and urgency of studying the novel mechanisms of the impaired wound healing and exploring now therapeutic options. MicroRNAs (miRNAs) have emerged as a novel class of noncoding RNAs with tremendous biological functions. The goals of this project are to determine the miRNA mechanism and therapeutics of impaired wound healing. Using RCR based array and deep-sequencing analysis as well as qRT-PCR, we identified a group of miRNAs that are aberrantly expressed in bone marrow (BM)-derived endothelial progenitor cells (EPCs) isolated from diabetic and aged individuals and mice. Among them, let-7 family members, let-7b, let-7c, let-7d, and let-7d* are significantly increased. The hypothesis of this proposal is that Let-7 family members play important roles in the impaired cellular functions of EPCs and impaired wound healing in diabetes and aged individuals via their target genes. miRNA-reprogrammed "smart" EPCs via modulators of let-7 family may represent a novel effective therapy for chronic wounds. We will test our hypothesis via three specific aims: Aim 1 will determine the effects of let-7 family members on the migration, proliferation and apoptosis of EPCs in vitro. Aim 2 will determine the effects of let-7-reprogrammed autologous EPCs on wound angiogenesis and wound healing in diabetic and aged mice in vivo. Aim 3 will determine the molecular mechanisms of let-7-mediated biological effects. The biological role of let-7 family on refractory wounds via gene reprogramming in EPCs is an unexplored research field. The proposed study will utilize the latest cellular, molecular, and whole animal approaches to target the major unsolved biomedical challenge: chronic wounds. Uncovering the novel molecular mechanisms and therapeutics of the impaired wound healing at translational level via miRNAs may open a new scientific paradigm in chronic wounds. PUBLIC HEALTH RELEVANCE: The biological role of let-7 family on refractory wound via gene reprogramming in endothelial progenitor cells (EPCs) is an unexplored research field. The goal of this project is to test our hypothesis: Let-7 family members play important roles in impaired cellular functions of EPCs and impaired wound healing via their target genes.