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Targeting vascular dysfunction to promote lung repair and fibrosis resolution

针对血管功能障碍促进肺修复和纤维化消退

基本信息

批准号：
10444342
负责人：
Giovanni Ligresti
金额：
$ 71.71万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-03-05 至 2026-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10444342
关键词：
ATAC-seq Ablation Address Aging Attenuated Binding Biochemical Bleomycin Blood Vessels Blood capillaries Capillary Endothelial Cell Cell Death ChIP-seq Chromatin Chronic Coculture Techniques Collagen Coupled Data Disease Disease Progression Endothelial Cells Endothelium Enhancers Epigenetic Process Exhibits Fibroblasts Fibrosis Foundations Functional disorder Gene Silencing Genes Genetic Genetic Transcription Goals Grant Histologic Histones Homeostasis Human Hypoxia Immunohistochemistry Impairment In Vitro Inflammation Inflammatory Inflammatory Response Knowledge Laboratories Link Lung Mediating Mediator of activation protein Methods Molecular Molecular Abnormality Mus Natural Immunity Pathway interactions Patients Pharmacology Phase Phenotype Play Production Pulmonary Fibrosis Reporter Resolution Role STAT1 gene Signal Transduction System Testing Therapeutic Transgenic Mice Vascular Diseases Vascular remodeling aged angiogenesis base cell age cell regeneration conditional knockout deep sequencing defined contribution design endothelial dysfunction endothelial stem cell enhancer binding protein experimental study gene function gene repair genetic signature genome-wide human model idiopathic pulmonary fibrosis in vivo Model inhibitor injured lung injury lung repair multiple omics novel overexpression paracrine progenitor repaired response to injury senescence single-cell RNA sequencing transcription factor transcriptome sequencing vascular abnormality vascular endothelial dysfunction

项目摘要

ABSTRACT Idiopathic Pulmonary Fibrosis (IPF) is a chronic, fatal disease of aging with limited therapeutic options. Despite IPF lungs feature vascular abnormalities, including capillary rarefaction and vascular leak, the impact of vascular endothelial dysfunction in the progression of this disease has remained unexplored. This proposal is designed to fill this knowledge gap. Multi-omics analysis of endothelial cells (ECs) from young and aged mouse lungs performed in our laboratory implicated the transcription factor ERG as an orchestrator of pulmonary vascular repair and inflammation, whose homeostatic function is impaired during fibrosis with aging. Genetic ablation of endothelial ERG in young mice led to increased inflammation, vascular rarefaction, and perpetuated lung fibrosis following bleomycin challenge mirroring the aged lung phenotype. ERG silencing in human lung ECs in vitro led to the increased secretion of fibrogenic mediators that promoted IPF-derived lung fibroblast activation. Whole lung scRNA-seq combined with FACS analysis revealed reduced number of lung progenitor ECs, known as general capillary (gCap) ECs, in ERG deficient mice compared to WT mice; this alteration was also observed in lungs derived from IPF patients. Pharmacologic inhibition of the enhancer-binding protein and epigenetic regulator BRD4 reversed inflammatory responses in ERG-silenced human lung ECs in vitro and restored gCap EC identity in IPF lung explants ex vivo. Moreover, IPF and bleomycin-induced mouse lung fibrosis were associated with overexpression of genes that regulate necroptosis, a form of programmed inflammatory cell death implicated in aging and fibrosis. In addition, inhibition of necroptosis with the specific inhibitor Necrostatin- 1 attenuated inflammation induced by ERG silencing in human lung ECs. Based on these findings we hypothesize that aging-associated epigenetic remodeling impairs ERG transcription in injured lung gCap ECs leading to dysregulated inflammation, capillary rarefaction, and persistent lung fibrosis. Aim 1 of this proposal will characterize the influence of ERG on lung gCap EC regeneration and fibrosis. Aim 2 will define the role of epigenetic mechanisms in lung endothelial ERG chromatin interaction, transcription, and vascular aging. Aim 3 will establish the contribution of endothelial necroptosis to lung vascular abnormalities and fibrosis in aged mice with compromised ERG functions. We will test our hypothesis using in vitro, ex vivo and in vivo models including an organotypic model of human IPF. gCap-enriched ECs and fibroblasts will be isolated from human IPF lungs. Endothelial fibrogenic activity will be evaluated in an endothelial-fibroblast co-culture systems. Conditional knockout and lineage tracing approaches will be used to investigate the role of ERG in endothelial homeostasis and lung fibrosis. Fibrosis, vascular leak, capillary rarefaction, and hypoxia will be evaluated with molecular, biochemical, and histological methods. Additional methods include immunohistochemistry, ChIP-seq analysis, and pharmacologic inhibition of selected targets. This proposal will define endothelial alterations that are critical in the progression of lung fibrosis toward our long-term goal of identifying novel targets for the treatment of IPF.

摘要