Defining the role of mechanoresponsive adipocyte-to-fibroblast transition in wound fibrosis.

定义机械反应性脂肪细胞向成纤维细胞转变在伤口纤维化中的作用。

• 批准号: 10654464
• 负责人: MICHAEL T LONGAKER
• 金额: $ 34.25万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2028-02-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10654464
• 关键词: Adipocytes; Adipose tissue; Adopted; Adult; Anatomy; Cells; Cicatrix; Clinical; Communication; Cutaneous; Data; Deposition; Dermal; Development; Dipeptidyl-Peptidase IV; Extracellular Matrix; Fibroblasts; Fibrosis; Flow Cytometry; Focal Adhesion Kinase 1; Genes; Genetic; Growth; Histologic; Histology; Human; Hypertrophic Cicatrix; Immunohistochemistry; In Situ; In Vitro; Injury; Knockout Mice; Machine Learning; Mechanics; Medical; Modeling; Molecular; Morbidity - disease rate; Mus; Myofibroblast; Natural regeneration; Pathway interactions; Phase; Phenotype; Piezo 1 ion channel; Piezo 2 ion channel; Population; Process; Proteomics; Reporting; Role; Signal Pathway; Signal Transduction; Skin; Societies; Surface; System; Time; Tissues; Transgenic Organisms; United States; Work; Wound models; Xenograft Model; adipocyte differentiation; cell type; cost; data integration; epigenomics; functional loss; healing; image processing; in vivo; mechanical drive; mechanical force; mechanical signal; mechanotransduction; molecular dynamics; molecular phenotype; mortality; mouse model; multimodal data; multiple omics; new therapeutic target; novel; novel therapeutics; prevent; profibrotic fibroblast; psychosocial; regenerative; response; skin fibrosis; skin regeneration; skin wound; skin xenograft; small molecule; subcutaneous; therapeutic target; transcriptomics; translational goal; treatment strategy; wound; wound care; wound environment; wound healing

7. Project Summary/Abstract Adult human skin heals by developing fibrotic scar tissue, which can result in devastating disfigurement, growth restriction, and permanent functional loss. Despite a plethora of clinical options, no current treatment strategies successfully prevent or reverse this fibrotic process, and scars and their sequelae cost the United States over $20 billion every year. Progress towards the development of new therapies has been significantly hindered by a lack of understanding of the cell populations responsible for scarring and their molecular dynamics. Studies in recent years have reported that adipocytes in wounds are capable of transitioning into fibroblasts (and vice versa); however, the extent to which adipocyte-to-fibroblast transition contributes to wound fibrosis (scarring), and whether this process can be targeted to prevent scarring, remain unknown. In this proposal, we explore for the first time the role of tissue mechanics in conversion of dermal adipocytes to scarring fibroblasts within the wound environment. First, employing genetic lineage tracing, we will use histology, immunohistochemistry, and flow cytometry to study adipocyte-to-fibroblast transition and to interrogate the molecular phenotype of adipocyte lineage-derived fibroblasts within wounds. Second, we will use a Rainbow mouse model to interrogate clonal dynamics of adipocyte-to-fibroblast transition in wounds, and will apply an integrated multi-omic analysis, with single-cell transcriptomic (scRNA-seq) and epigenomic (scATAC-seq), spatial transcriptomic (Visium) and proteomic (CODEX), and quantitative extracellular matrix (ECM) ultrastructural analyses, in order to robustly define the molecular drivers and pathways involved in adipocyte-to-fibroblast conversion during scarring. Third, as our preliminary data strongly support a mechanotransduction mechanism underlying adipocyte-to-fibroblast transition during wound healing, we will inhibit mechanical signaling in adipocytes using both small molecule and transgenic approaches in order to block adipocyte-to-fibroblast transition. We will apply a similar multi-omic analysis to elucidate the molecular dynamics that differentiate wound adipocyte dynamics in the context of intact versus blocked mechanical signaling and determine how inhibiting mechanically driven adipocyte-to-fibroblast conversion may reduce fibrosis and yield wound regeneration. Our ultimate translational goal is to develop therapeutics that target fibrogenic wound cell dynamics to promote regenerative healing. Collectively, the proposed work will significantly enhance our understanding of the key molecular and cellular determinants of cutaneous scarring, inform the development of novel anti-scarring therapies, and shed light on the contributions of adipose tissue to wound fibrosis.

7。项目摘要/摘要 成人人的皮肤通过发展纤维化疤痕组织来治愈，这可能导致毁灭性，生长 限制和永久功能损失。尽管有很多临床选择，但没有当前的治疗策略 成功防止或扭转这一纤维化过程，疤痕及其后遗症使美国付出了代价 每年为200亿美元。朝着开发新疗法的进展受到了极大的阻碍 缺乏对负责疤痕及其分子动力学的细胞群的了解。研究 近年来报道说，伤口中的脂肪细胞能够过渡到成纤维细胞（和恶化 versa）;但是，脂肪细胞到成纤维细胞过渡的程度有助于伤口纤维化（疤痕）， 以及是否可以针对此过程以防止疤痕，仍然未知。在此提案中，我们探索 组织力学在真皮脂肪细胞转化中的作用第一次 伤口环境。首先，采用遗传谱系追踪，我们将使用组织学，免疫组织化学和 流式细胞仪研究脂肪细胞至纤维细胞过渡并询问脂肪细胞的分子表型 谱系衍生的成纤维细胞在伤口内。其次，我们将使用彩虹鼠标模型来询问克隆 伤口中脂肪细胞到纤维细胞过渡的动力学，并将应用综合的多摩尼克分析 单细胞转录组（SCRNA-SEQ）和表观基因组（SCATAC-SEQ），空间转录组（visium）和 蛋白质组学（法典）和定量细胞外基质（ECM）超微结构分析，以便于 定义疤痕过程中脂肪细胞到纤维细胞转换涉及的分子驱动因素和途径。第三， 因为我们的初步数据强烈支持脂肪细胞至纤维细胞的机械转移机制 在伤口愈合期间的过渡，我们将使用小分子和 转基因方法为了阻止脂肪细胞至纤维细胞过渡。我们将应用类似的多摩尼克 分析以阐明在完整背景下区分伤口脂肪细胞动力学的分子动力学 与阻塞的机械信号传导，并确定如何抑制机械驱动的脂肪细胞至纤维细胞 转化可以减少纤维化并产生伤口再生。我们的最终翻译目标是发展 靶向纤维化伤口细胞动力学以促进再生愈合的疗法。集体， 拟议的工作将显着增强我们对关键分子和细胞决定因素的理解 皮肤疤痕，告知新型反施加疗法的发展，并阐明贡献 脂肪组织的纤维化伤。