MFAP5 and Skin Scar Formation

项目总结 这项申请提出了一个定制的研究培训计划，旨在促进 申请人成为一名独立的调查员。该计划包括生物信息学和 实验室实验，以及量身定做的专业和职业发展机会。培训 该计划得到了UIC现有的当地和机构资源的支持。建议数 研究将检查控制疤痕形成的机制，疤痕形成是愈合反应的常见结果。在……里面 成年人，伤口修复的结果几乎总是由无序的细胞外组成的纤维疤痕 矩阵(ECM)。尽管瘢痕形成的调控很复杂，但一个关键特征是成纤维细胞(FB)的激活。 这会产生ECM和收缩力。疤痕形成和纤维化发生在许多组织中，并可能导致 受影响的器官系统严重受损。我们实验室最近的研究发现了一种新的FB 在伤口中的作用可能与疤痕形成有关。这些研究表明，创伤的FBS可以作为非 专业吞噬细胞和吞噬凋亡细胞。随着凋亡细胞的吞噬，FBS发展为纤维化 表型迁移增强，收缩能力增强(α-SMA表达)，胶原增加 综合。纤维吞噬细胞性FBS中显著上调的一个因素是微纤维相关 蛋白5(MFAP5或微原纤维相关糖蛋白2/MAGP2)。MFAP5影响微原纤维功能和 可以调节细胞信号通路。有趣的是，MFAP5与几个人的肝纤维化有关 疾病，包括一些癌症和纤维性疾病。然而，人们对它在伤口愈合中的作用知之甚少。 和疤痕的形成。因此，本研究的目的是为了更好地了解MFAP5的功能 在皮肤愈合和疤痕形成方面。我们假设MFAP5在愈合的皮肤伤口中表达上调 改变FB反应，促进疤痕形成。在本研究中，MFAP5在伤口愈合中的作用 将在Mfap5-/-小鼠身上进行研究，Mfap5-/-小鼠是一种成熟的Mfap5基因敲除小鼠系。MFAP5对大鼠血管内皮细胞生长的影响 Fb的表型将在体外进一步研究。这项提议将利用体内和体外伤口愈合 分析和先进的生物信息学技术。目标1将评估MFAP5的缺失对伤口的影响 愈合，包括瘢痕胶原含量和结构，伤口破裂强度，肌成纤维细胞激活， 血管生成和伤口闭合。单细胞RNA测序将用于鉴定Fb亚群 在伤口愈合过程中产生MFAP5。Aim 2将研究MFAP5如何影响Fb功能和基因 利用体外伤口愈合试验、批量RNA测序和功能途径进行表达 分析。总之，这些目标将使人们更好地理解MFAP5在皮肤愈合中的重要性 伤口和疤痕的形成。从拟议的研究中获得的信息可能会导致 治疗纤维化疾病的新疗法和/或预后生物标记物的发现。

PROJECT SUMMARY This application proposes a customized research training plan designed to promote the development of the applicant into an independent investigator. The plan includes advanced training in both bioinformatics and laboratory experimentation, along with tailored professional and career development opportunities. The training plan is supported by the outstanding availability of local and institutional resources at UIC. The proposed research will examine the mechanisms that control scar formation, a common result of the healing response. In adults, the outcome of wound repair is almost always a fibrous scar composed of disorganized extracellular matrix (ECM). Although regulation of scar formation is complex, a key feature is fibroblast (FB) activation, which generates ECM and contractile forces. Scarring and fibrosis occurs in many tissues and can cause significant impairments of the organ system affected. Recent studies in our lab have identified a novel FB function in wounds that may be linked to scar formation. These studies show that wound FBs can act as non- professional phagocytes and ingest apoptotic cells. Following apoptotic cell engulfment, FBs develop a fibrotic phenotype with enhanced migration, increased contractility (α-SMA expression), and increased collagen synthesis. One factor found to be significantly upregulated in fibrotic phagocytic FBs is microfibril-associated protein 5 (MFAP5 or microfibril-associated glycoprotein 2/MAGP2). MFAP5 influences microfibril function and can regulate cell signaling pathways. Interestingly, MFAP5 has been linked to fibrosis in several human diseases, including some cancers and fibrotic diseases. Still, little is known regarding its role in wound healing and scar formation. Therefore, the goal of this study is to gain a better understanding of the function of MFAP5 in skin healing and scar formation. We hypothesize that upregulation of MFAP5 in the healing skin wound modifies the FB response and promotes scar formation. In this study, the role of MFAP5 in wound healing will be investigated in Mfap5-/- mice, a well-established Mfap5 knock out mouse line. The effect of MFAP5 on FB phenotype will be further investigated in vitro. This proposal will utilize in vivo and in vitro wound healing assays and advanced bioinformatics techniques. Aim 1 will assess how the loss of MFAP5 affects wound healing, including scar collagen content and architecture, wound breaking strength, myofibroblast activation, angiogenesis, and wound closure. Single-cell RNA sequencing will be used to identify the FB subpopulation that produces MFAP5 during wound healing. Aim 2 will examine how MFAP5 influences FB function and gene expression by utilizing in vitro wound healing assays as well as bulk RNA-sequencing and functional pathway analysis. Together, the Aims will lead to a better understanding of the importance of MFAP5 in healing skin wounds and scar formation. Information gained from the proposed research may lead to the development of novel therapeutics and/or discovery of a prognostic biomarker for treatment of fibrotic diseases.