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Targeting CEBPA To Restore Epithelial-Mesenchymal Homeostasis In Lung Fibrosis

靶向 CEBPA 恢复肺纤维化的上皮间质稳态

基本信息

批准号：
10815455
负责人：
Qi Tan
金额：
$ 47.03万
依托单位：
UNIVERSITY OF MINNESOTA
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：
2020-07-15 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10815455
关键词：
Apoptosis Attenuated BMP4 Bleomycin CEBPA gene CRISPR-mediated transcriptional activation Cell Communication Cell Proliferation Cells Chronic lung disease Cicatrix Coculture Techniques Cytoprotection Data Deposition Disease Disease Progression Epithelial Cells Epithelium Extracellular Matrix Fibroblasts Fibrosis Functional disorder Gene Expression Profiling Genes Genetic Transcription Goals Homeostasis Human Injury Knock-out Link Lung Mediating Mediator Mesenchymal Mesenchyme Molecular Mus Myofibroblast Organoids Outcome Pathologic Pathway interactions Patients Phenotype Process Publications Pulmonary Fibrosis Recombinant Proteins Resolution Role Signal Transduction Sorting Testing Therapeutic Therapeutic Intervention Transcriptional Regulation aged alveolar epithelium antagonist cell injury experimental study gain of function idiopathic pulmonary fibrosis in vivo insight loss of function lung development lung repair migration mouse model novel programs protein expression pulmonary function regenerative therapy repaired research study single-cell RNA sequencing targeted treatment transcriptome sequencing

项目摘要

PROJECT SUMMARY While tremendous progress has been made to understand mechanisms of epithelial cell injury and fibroblast activation in fibrosis, significant challenges remain in fully understanding the mechanisms of epithelial– mesenchymal crosstalk in normal lung homeostasis and during injury, repair and fibrosis. As in normal lung development, lung repair requires epithelial cells and mesenchymal cells to coordinate with each other to tune cell proliferation, migration, differentiation and apoptosis. Similarly, extracellular matrix (ECM) remodeling is mediated by a myriad of pro-fibrotic and anti-fibrotic factors that are precisely orchestrated by epithelium and mesenchyme during injury, repair and resolution. Therefore, aberrant epithelial mesenchymal interactions can lead to non-healing processes in lung repair with pathological scar formation due to myofibroblast accumulation and ECM deposition, ultimately contributing to pulmonary fibrosis. The long-term goal of this proposal is to restore epithelial-mesenchymal homeostasis in pulmonary fibrosis with transcriptional regulation of critical anti-fibrotic factors. Targeting mechanisms that augment the endogenous anti-fibrotic or fibrosis resolution signals during disease progression may serve as an attractive therapeutic strategy to alleviate lung fibrosis and restore lung function. Recently, single-cell and bulk RNA sequencing identified loss of normal epithelial cell identities and gain of abnormal indeterminate states of differentiation in IPF, with CEBPA identified as a key transcriptional regulator diminished in IPF that was common to multiple independent studies. Our central hypothesis is that CEBPA deficiency in lung epithelial cells promotes lung fibroblast activation and fibrosis, that it normally increases during repair to resolve fibrosis, and that it can be therapeutically restored using non-genome editing CRISPR gene activation to promote fibrosis resolution. This hypothesis will be tested with three specific aims: First, we will evaluate the role of Cebpa signaling from epithelial cells in maintaining lung epithelial-mesenchymal homeostasis and its protective role in pulmonary fibrosis. Second, we will determine whether increasing Cebpa expression with CRISPR gene activation is able to restore epithelial-mesenchymal homeostasis and attenuate fibroblast activation and fibrosis. We will develop and optimize an AAV-mediated approach to enhance Cebpa expression using non-genome editing CRISPR activation in the lung of aged mice that display non-resolving lung fibrosis after bleomycin injury. Lastly, We will test whether the anti-fibrotic effect of epithelial Cebpa expression is mediated by BMP4 and perform RNA-seq- based gene expression analysis of sorted epithelial cells from Cebpa gain and loss of function studies to identify additional novel candidate mediators of epithelial-mesenchymal homeostasis. Taken together, the proposed research studies will reveal critical anti-fibrotic targets for therapeutic interventions aimed at restoring epithelial-mesenchymal homeostasis and alleviating pulmonary fibrosis.

项目摘要虽然在了解上皮细胞损伤和成纤维细胞损伤的机制方面取得了巨大进展，激活的纤维化，重大的挑战仍然是在充分理解上皮细胞的机制，在正常肺稳态和损伤、修复和纤维化期间的间充质串扰。就像正常的肺一样在发育过程中，肺的修复需要上皮细胞和间充质细胞相互协调来调节，细胞增殖、迁移、分化和凋亡。类似地，细胞外基质（ECM）重塑是一种细胞外基质重塑。由无数的促纤维化和抗纤维化因子介导，这些因子由上皮细胞精确地协调，间充质在损伤，修复和决议。因此，异常的上皮间质相互作用可以导致肺修复中不愈合过程，具有由肌成纤维细胞引起的病理性瘢痕形成累积和ECM沉积，最终导致肺纤维化。长期目标是一项通过转录调控恢复肺纤维化中上皮-间质稳态的建议关键的抗纤维化因子增强内源性抗纤维化或纤维化的靶向机制在疾病进展期间的分辨率信号可以作为一种有吸引力的治疗策略来减轻肺损伤。纤维化和恢复肺功能。最近，单细胞和批量RNA测序确定了正常的缺失， CEBPA治疗IPF患者的上皮细胞特性和异常不确定分化状态的获得在IPF中被鉴定为一种关键的转录调节因子，在多个独立的问题研究我们的中心假设是CEBPA在肺上皮细胞中的缺乏促进肺成纤维细胞激活和纤维化，它通常在修复过程中增加以解决纤维化，并且它可以是使用非基因组编辑CRISPR基因激活以促进纤维化消退来治疗性恢复。这我们将通过三个具体目标来检验这一假设：首先，我们将评估Cebpa信号在以下方面的作用：上皮细胞在维持肺上皮-间质平衡中的作用及其对肺损伤的保护作用纤维化其次，我们将确定CRISPR基因激活是否能够增加Cebpa表达，以恢复上皮-间充质稳态并减弱成纤维细胞活化和纤维化。我们将开发并使用非基因组编辑CRISPR优化AAV介导的方法以增强Cebpa表达在博来霉素损伤后显示出未消退的肺纤维化的老年小鼠的肺中的活化。最后，我们将测试上皮Cebpa表达的抗纤维化作用是否由BMP 4介导，并进行RNA-seq- 对来自Cebpa获得和丧失功能研究的分选上皮细胞进行基于基因表达的分析，鉴定上皮-间充质稳态的其它新候选介质。综合起来，拟议的研究将揭示关键的抗纤维化目标的治疗干预，旨在恢复上皮-间质稳态和减轻肺纤维化。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Transcriptional analysis of lung fibroblasts identifies PIM1 signaling as a driver of aging-associated persistent fibrosis.

DOI：
10.1172/jci.insight.153672
发表时间：
2022-03-22
期刊：
JCI insight
影响因子：
8
作者：
Pham TX;Lee J;Guan J;Caporarello N;Meridew JA;Jones DL;Tan Q;Huang SK;Tschumperlin DJ;Ligresti G
通讯作者：
Ligresti G

Dopamine receptor signaling regulates fibrotic activation of retinal pigmented epithelial cells.

多巴胺受体信号传导调节视网膜色素上皮细胞的纤维化活化。