MFAP5 and Skin Scar Formation

MFAP5 和皮肤疤痕形成

• 批准号: 10605379
• 负责人: Chen Han
• 金额: $ 5.27万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-02 至 2027-05-01
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10605379
• 关键词: Adult; Affect; Apoptotic; Architecture; Area; Bioinformatics; Biological Assay; Cells; Chemoresistance; Cicatrix; Collagen; Complex; Contracture; Data; Development; Disease; Esthetics; Extracellular Matrix; Fibroblasts; Fibrosis; Gene Expression; Genes; Genomics; Goals; Hemostatic function; Human; Hypertrophic Cicatrix; Impairment; In Vitro; Inflammation; Inflammatory; Ingestion; Institution; Keloid; Knockout Mice; Lead; Link; Liver Cirrhosis; MFAP1 gene; Malignant Neoplasms; Mediator; Microfibrils; Mus; Myocardial Infarction; Myofibroblast; Ontology; Organ; Outcome; Pain; Pathogenesis; Pathologic; Pathway Analysis; Pathway interactions; Patients; Phagocytes; Phase; Phenotype; Process; Production; Prognostic Marker; Proliferating; Proteins; Pulmonary Fibrosis; RNA; Regulation; Research; Research Personnel; Research Training; Resources; Role; Signal Pathway; Signal Transduction; Skin; Skin wound healing; Source; Surface; Systemic Scleroderma; Techniques; Tissues; Training; Transfection; Up-Regulation; Width; Work; angiogenesis; body system; career development; cell motility; comparison control; design; differential expression; experimental study; extracellular; fibrillin; healing; human disease; idiopathic pulmonary fibrosis; in vivo; laboratory experiment; migration; novel; novel therapeutics; overexpression; recruit; repair function; response; single-cell RNA sequencing; skin fibrosis; skin regeneration; targeted biomarker; therapeutic biomarker; tissue injury; tissue repair; transcriptome; transcriptome sequencing; vector; wound; wound closure; wound healing

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.

项目概要 该申请提出了一个定制的研究培训计划，旨在促进该领域的发展 申请人成为独立调查员。该计划包括生物信息学和 实验室实验，以及量身定制的专业和职业发展机会。培训内容 该计划得到了伊利诺伊大学芝加哥分校当地和机构资源的支持。拟议的 研究将检查控制疤痕形成的机制，这是愈合反应的常见结果。在 成年人，伤口修复的结果几乎总是由杂乱的细胞外物质组成的纤维疤痕 矩阵（ECM）。尽管疤痕形成的调节很复杂，但一个关键特征是成纤维细胞 (FB) 激活， 它产生 ECM 和收缩力。许多组织中都会出现疤痕和纤维化，并可能导致 受影响的器官系统明显受损。我们实验室最近的研究发现了一种新型 FB 在可能与疤痕形成有关的伤口中发挥作用。这些研究表明，伤口 FB 可以充当非 专业吞噬细胞并吞噬凋亡细胞。凋亡细胞吞噬后，FBs 形成纤维化 具有增强迁移、增加收缩性（α-SMA 表达）和增加胶原蛋白的表型 合成。发现纤维化吞噬 FB 中显着上调的一个因素是微纤维相关的 蛋白 5（MFAP5 或微原纤维相关糖蛋白 2/MAGP2）。 MFAP5 影响微纤维功能 可以调节细胞信号通路。有趣的是，MFAP5 与多种人类的纤维化有关 疾病，包括一些癌症和纤维化疾病。尽管如此，人们对其在伤口愈合中的作用仍知之甚少 和疤痕形成。因此，本研究的目的是更好地了解 MFAP5 的功能 皮肤愈合和疤痕形成。我们假设 MFAP5 在皮肤伤口愈合过程中上调 改变 FB 反应并促进疤痕形成。在这项研究中，MFAP5 在伤口愈合中的作用 将在 Mfap5-/- 小鼠（一种成熟的 Mfap5 敲除小鼠系）中进行研究。 MFAP5对的影响 FB表型将在体外进一步研究。该提案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 的丢失如何影响伤口 愈合，包括疤痕胶原蛋白含量和结构、伤口断裂强度、肌成纤维细胞活化、 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。目标 2 将研究 MFAP5 如何影响 FB 功能和基因 利用体外伤口愈合测定以及批量 RNA 测序和功能途径进行表达 分析。这些目标将共同促进人们更好地了解 MFAP5 在治愈皮肤方面的重要性 伤口和疤痕形成。从拟议的研究中获得的信息可能会导致开发 用于治疗纤维化疾病的新疗法和/或预后生物标志物的发现。