Mechanoresponsive Engrailed-1-negative fibroblasts activate Engrailed-1 to promote fibrosis in wound healing

机械反应性 Engrailed-1 阴性成纤维细胞激活 Engrailed-1 以促进伤口愈合中的纤维化

基本信息

批准号：
10550197
负责人：
MICHAEL T LONGAKER
金额：
$ 31.76万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-01 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10550197
关键词：
3-Dimensional Adult Algorithms Anatomic Surface Behavior Categories Cells Cellular Assay Characteristics Chemicals Chromatin Cicatrix Clinical Collagen Connective Tissue Cues Cultured Cells Cutaneous Data Dermal Dermis Development Dipeptidyl-Peptidase IV Dorsal Elements Engraftment Environment Extracellular Matrix Fiber Fibroblasts Fibrosis Fluorescence-Activated Cell Sorting Focal Adhesion Kinase 1 Genetic Transcription Goals Growth Hair follicle structure High-Throughput Nucleotide Sequencing Histologic Histology Hydrogels Immunohistochemistry In Vitro Individual Injury Maps Measures Mechanical Stress Mechanics Mediating Medical Microscopy Molecular Morbidity - disease rate Mus Outcome Papillary Pathway interactions Population Pregnancy Process Production Protein Inhibition Proteins Reporting Role Signal Pathway Signal Transduction Site Skin Societies Specimen Subcutaneous Tissue Surface Tamoxifen Time Tissues Transgenic Mice Transposase United States Verteporfin Visual Wild Type Mouse Work analog cost digital epigenomic profiling epigenomics experimental study fetal functional loss healing in vivo in vivo Model inhibitor machine learning algorithm mechanical force mechanical signal mechanotransduction mortality mouse model novel novel therapeutics postnatal prevent reconstruction regenerative response single-cell RNA sequencing skin regeneration skin wound therapeutic development therapeutic target tissue culture tool transcriptomic profiling transcriptomics translational goal treatment strategy wound wound bed wound environment wound healing

项目摘要

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 specific cell populations responsible for scarring. In 2015, our group reported that Engrailed-1 (En-1) lineage-positive fibroblasts (EPFs) are responsible for the vast majority of dorsal scar production in postnatal mice. In early fetal gestation, mice heal scarlessly via skin regeneration, an ideal outcome mediated by En-1 lineage-negative fibroblasts (ENFs; the predominant fetal fibroblast). However, it has not been established if ENFs contribute to postnatal wound healing. In this proposal, we explore for the first time the postnatal conversion of ENFs to pro-fibrotic EPFs (postnatally-derived EPFs; pEPFs) within the wound environment. First, histology, immunohistochemistry, and wounding in a novel transgenic mouse model will be used to study the conversion of ENFs to pEPFs during wound healing. By examining the behavior of ENF subpopulations (derived from papillary dermis, reticular dermis, and hypodermis) in the wound environment and confirming our findings in a tamoxifen-inducible mouse model of En-1 activation, we will precisely define the ENF population that gives rise to pro-fibrotic pEPFs. Second, we will establish the specific wound environment cues that drive ENF-to-EPF transition. Given that mechanical forces are known to modulate both scar burden and fibroblast activity, we will use in vitro and in vivo models to examine the effects of mechanical environment on En-1 activation. We will further use transcriptomic and epigenomic profiling to explore the role of mechanotransduction signaling in ENF-to-EPF transition and pEPF function. Third, having established a mechanotransduction mechanism underlying En-1 activation in wound ENFs, we will inhibit mechanotransduction signaling with the goal of blocking ENF-to-EPF transition. Specifically, we will assess whether blocking mechanotransduction results in ENF-mediated wound healing with reduced fibrosis. Our ultimate translational goal is to develop therapeutics that target fibrogenic fibroblasts 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 cellular origin of dermal scarring fibroblasts.

7. 项目总结/摘要成人皮肤通过形成纤维化疤痕组织来愈合，这可能导致毁灭性的毁容、生长限制和永久性功能丧失。尽管临床选择众多，但目前尚无治疗策略成功地阻止或逆转这种纤维化过程，而疤痕及其后遗症使美国付出了超过每年200亿美元。新疗法的开发进展受到了严重阻碍缺乏对造成疤痕的特定细胞群的了解。 2015年，我们小组报告称 Engrailed-1 (En-1) 谱系阳性成纤维细胞 (EPF) 是造成绝大多数背侧疤痕的原因产后小鼠的生产。在胎儿妊娠早期，小鼠通过皮肤再生实现无疤痕愈合，这是一个理想的结果由 En-1 谱系阴性成纤维细胞（ENF；主要胎儿成纤维细胞）介导。然而，它并没有被确定 ENF 是否有助于产后伤口愈合。在本提案中，我们首次探讨了出生后伤口内 ENF 转化为促纤维化 EPF（出生后衍生的 EPF；pEPF）环境。首先，将研究新型转基因小鼠模型中的组织学、免疫组织化学和损伤。用于研究伤口愈合过程中 ENF 向 pEPF 的转化。通过检查 ENF 的行为伤口环境中的亚群（源自乳头状真皮、网状真皮和皮下组织）为了证实我们在他莫昔芬诱导的 En-1 激活小鼠模型中的发现，我们将精确定义 ENF 产生促纤维化 pEPF 的人群。其次，我们将建立特定的伤口环境线索推动 ENF 到 EPF 的转变。鉴于已知机械力可以调节疤痕负担和成纤维细胞活性，我们将使用体外和体内模型来检查机械环境对成纤维细胞活性的影响 En-1激活。我们将进一步利用转录组和表观基因组分析来探索 ENF 到 EPF 转换和 pEPF 功能中的机械转导信号。三、建立了伤口 ENF 中 En-1 激活的机械转导机制，我们将抑制力传导信号传导的目的是阻止 ENF 到 EPF 的转变。具体来说，我们将评估阻断机械传导是否会导致 ENF 介导的伤口愈合并减少纤维化。我们的最终的转化目标是开发针对纤维化成纤维细胞以促进再生的治疗方法康复。总的来说，拟议的工作将显着增强我们对关键分子和皮肤疤痕的细胞决定因素，为新型抗疤痕疗法的开发提供信息，并摆脱揭示真皮疤痕成纤维细胞的细胞起源。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Bioprinted Hydrogels for Fibrosis and Wound Healing: Treatment and Modeling.

DOI：
10.3390/gels9010019
发表时间：
2022-12-27
期刊：
Gels (Basel, Switzerland)
影响因子：
0
作者：
通讯作者：

Desmoplastic stromal signatures predict patient outcomes in pancreatic ductal adenocarcinoma.

DOI：
10.1016/j.xcrm.2023.101248
发表时间：
2023-11-21
期刊：
CELL REPORTS MEDICINE
影响因子：
14.3
作者：
Mascharak, Shamik;Guo, Jason L.;Foster, Deshka S.;Khan, Anum;Davitt, Michael F.;Nguyen, Alan T.;Burcham, Austin R.;Chinta, Malini S.;Guardino, Nicholas J.;Griffin, Michelle;Lopez, David M.;Miller, Elisabeth;Januszyk, Michael;Raghavan, Shyam S.;Longacre, Teri A.;Delitto, Daniel J.;Norton, Jeffrey A.;Longaker, Michael T.
通讯作者：
Longaker, Michael T.