Damage-Associated Molecular Patterns Driving Fibrosis Progression in Scleroderma

驱动硬皮病纤维化进展的损伤相关分子模式

基本信息

批准号：
10328406
负责人：
John Varga
金额：
$ 46.33万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-01-13 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10328406
关键词：
3-Dimensional Address Affect Automobile Driving Bacterial Artificial Chromosomes Biological Markers Biology Biopsy Cells Characteristics Chronic Clinical Cutaneous sclerosis Disease Disease Progression Disease model Enzymes Experimental Models Extracellular Matrix Failure Fibroblasts Fibrosis Genetic Glycoproteins Goals Human Immune Immunologic Receptors Impairment In Vitro Individual Inflammation Laboratories Ligands Light Link Longitudinal cohort Lung Maps Measures Mediating Mediator of activation protein Modeling Molecular Mus Myeloid Cells Myofibroblast Natural Immunity Organ Organoids Pathogenesis Pathogenicity Pathway interactions Patients Pattern Pattern recognition receptor Phenocopy Play Population Process Production Progressive Disease Proteomics Publishing Regulation Role Sampling Scleroderma Severity of illness Signal Pathway Signal Transduction Skin Source Specificity Stable Disease Stimulus Stromal Cells Systemic Scleroderma TLR4 gene Technology Tenascin Testing Tissues Transforming Growth Factor beta Transgenic Mice Ubiquitin Variant Work antifibrotic treatment cell injury cell type clinically significant druggable target effective therapy genomic locus guided inquiry human disease in vivo inhibitor/antagonist insight multidisciplinary novel novel marker novel therapeutics programs response risk variant single cell sequencing single-cell RNA sequencing skin fibrosis therapeutic target transcriptomics

项目摘要

June 3 2018 ABSTRACT Synchronous fibrosis in multiple organs is the defining hallmark of systemic sclerosis (SSc), but its pathogenesis remains poorly understood, and there is an urgent need to discover “druggable” targets. The pattern recognition receptor Toll-like receptor 4 (TLR4), a vital mediator of innate immunity, is expressed on both immune and stromal cells, and can be activated by endogenous ligands called “damage-associated molecular patterns” (DAMPs). Published and preliminary work from our laboratories show that TLR4 and its endogenous ligand tenascin-C are likely to play important roles in multi-organ fibrosis in SSc. Notably, tenascin-C itself elicits core fibrotic responses including ECM production and matrix stiffening in fibroblasts and 3D human skin organoid models. TLR4 activity is regulated by the ubiquitin-editing enzyme A20, which is a major risk gene for SSc. However, the mechanism linking SSc-associated A20 variants and pathogenesis are unknown. We demonstrated that A20 expression is reduced in SSc skin biopsies. Surprisingly, we found that A20 inhibited core fibrotic responses, and mice that are haploinsufficient for A20 showed markedly aggravated non- inflammatory skin fibrosis. We hypothesize that persistence of fibrosis in SSc could be explained by activated TLR4 signaling that is triggered by tenascin-C and other DAMPs, and chronically sustained by impaired A20 function. The cell types with increased profibrotic TLR4 pathway activity, and the ensemble of profibrotic DAMPs and specific domains, remain unknown. Moreover, the clinical correlates of reduced A20 in SSc, cell type-specific regulation and anti-fibrotic activity of A20, and the mechanisms linking reduced A20 and SSc pathogenesis, have never been investigated. To address these critical gaps, Aim 1 will determine cell type- and stimulus-specific roles and mechanisms of DAMP-TLR4 signaling in two separate models of experimentally-induced multi-organ fibrosis; map key tenascin-C domains and identify additional DAMPs as potential SSc biomarkers and therapeutic targets; Aim 2 will define the clinical correlates of A20 expression in a longitudinal cohort of SSc patients; and define distinct functions of A20 in the fibrotic process using novel A20-deficient and A20 humanized BAC transgenic mice. Aim 3 will determine the cellular sources and function of TLR4 signaling pathway activity in skin and lung from SSc patients and controls. Employing human disease samples from the established Northwestern and Yale Scleroderma Programs, combined with in vitro and in vivo disease models and state-of-the-art technologies including comprehensive matrisome analysis and unbiased single cell RNA sequencing of multiple tissue, our investigative team is poised to generate notable advances in understanding TLR4 signaling in SSc. The information in turn will guide discovery of novel biomarkers and therapies.

2018年6月3日抽象的多个器官的同步纤维化是系统性硬化症的定义标志（SSC），但它发病机理仍然很少了解，并且迫切需要发现“可吸毒”靶标。模式识别受体Toll样受体4（TLR4）是先天免疫的重要介体，是在免疫细胞和基质细胞上同时表达，可以通过称为的内源配体激活 “损伤相关的分子模式”（湿）。从我们的实验室表明，TLR4及其内源性配体Tenascin-C可能在 SSC中的多器官纤维化。值得注意的是，Tenascin-C本身引发了包括ECM在内的核心纤维化反应成纤维细胞和3D人类皮肤手机体模型中的生产和基质僵硬。 TLR4活性是由泛素编辑酶A20调节，这是SSC的主要风险基因。但是，连接SSC相关A20变体和发病机理的机制尚不清楚。我们证明了 SSC皮肤活检中的A20表达降低。令人惊讶的是，我们发现A20抑制了核心纤维化反应和对A20的单倍弹性的小鼠显着汇总的非 - 炎症性皮肤纤维化。我们假设SSC中纤维化的持久性可以通过激活的TLR4信号传导由Tenascin-C和其他潮湿触发，并长期持续通过受损的A20功能。纤维化TLR4途径活性增加的细胞类型，并且纤维化潮湿和特定域的集合仍然未知。而且，临床 SSC中A20降低的相关性，细胞类型特异性调节和A20的抗纤维化活性以及连接减少A20和SSC发病机理的机制从未进行过研究。解决这些关键的差距，AIM 1将确定细胞类型和刺激特异性角色和机制在实验诱导的多器官纤维化的两个单独模型中，潮湿-TLR4信号传导；地图关键的Tenascin-C结构域并确定其他潮湿是潜在的SSC生物标志物和治疗目标； AIM 2将定义SSC纵向队列中A20表达的临床相关性患者;并使用新型A20缺陷和A20定义A20在纤维化过程中的不同功能人源化的BAC转基因小鼠。 AIM 3将确定TLR4的细胞源和功能 SSC患者和对照组的皮肤和肺中的信号通路活动。采用人类疾病来自已建立的西北和耶鲁硬皮病计划的样品，并结合体外以及体内疾病模型和最先进的技术，包括综合基质组分析和无偏的单细胞RNA测序多组织，我们的调查团队中毒在理解SSC中TLR4信号传导方面产生显着的进步。信息依次将指导新型生物标志物和疗法的发现。