Interstitial resident fibroblasts direct alveolar epithelial differentiation

间质驻留成纤维细胞直接肺泡上皮分化

• 批准号: 9235745
• 负责人: Anne-Karina Theresia Perl
• 金额: $ 47.58万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-01 至 2020-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9235745
• 关键词: Age; Alveolar; Binding; Binding Sites; Cell Communication; Cell Differentiation process; ChIP-seq; Chronic lung disease; Computer Analysis; Confocal Microscopy; Data; Deposition; Development; EZH2 gene; Elastin; Epigenetic Process; Epithelial; Epithelial Cells; Epithelium; Exhibits; Fibroblasts; Fibrosis; Flow Cytometry; Gene Expression; Genes; Genetic Transcription; Goals; Hamman-Rich syndrome; Histology; Histones; Homeostasis; Human; Impairment; In Vitro; Knowledge; Link; Lung; Lung diseases; Magnetic Resonance Imaging; Mesenchymal; Mesenchymal Stem Cells; Messenger RNA; Methyltransferase; Modification; Molecular; Mus; Myofibroblast; Natural regeneration; Nature; Organoids; Outcome; Pathway interactions; Pharmacology; Phenotype; Platelet-Derived Growth Factor alpha Receptor; Population; Pulmonary Fibrosis; Regulation; Regulator Genes; Research; Research Project Grants; Role; Signal Transduction; Smooth Muscle Actin Staining Method; Surface; Testing; Transgenic Mice; age related; aged; base; experimental study; genetic signature; histone methylation; improved; in vivo; inhibitor/antagonist; insight; interstitial; loss of function; lung development; lung injury; lung regeneration; lung repair; morphometry; novel therapeutics; paracrine; progenitor; programs; promoter; regenerative therapy; repaired; response; tissue repair; transcription factor; transcriptome sequencing

PROJECT SUMMARY: While tremendous progress has been made to understand the roles of epithelial pro- genitors in tissue repair and fibrotic mechanisms, little is known about the origin, nature of interaction and gene regulatory programs of fibroblast progenitors. Pulmonary research has been challenged by the remarkable complexity and diversity of fibroblast populations and mesenchymal stem cell populations. Our long-term goal is to understand the complexity of fibroblast populations and to identify their regulatory role on the epithelium during alveolarization and epithelial repair. Our primary objective is to identify key regulators of fibroblast dif- ferentiation in the lung. The present application is based on our preliminary data that identifies GATA6 as a key transcription factor in matrix iReFs, which are indispensable for alveolarization. Our central hypothesis is that GATA6 regulates gene expression in iReFs that sub sequentially induce matrix iReF specification and para- crine interactions with AECs during development and repair. The rationale for the proposed research is that understanding the regulation of fibroblast subpopulations, and their distinct roles in epithelial-mesenchymal cell interactions, will provide new answers for fundamental questions regarding lung homeostasis and repair. This hypothesis will be tested with three specific aims: 1) that matrix iReFs instruct AEC1 differentiation. 2) that GATA6 transcriptionally regulates a set of matrix fibroblast signature genes and 3) that H3K27me silencing marks regulate GATA6 and matrix gene expression in aged iReFs. In the first aim, alveolosphere cultures will be used to interrogate the epithelial-mesenchymal crosstalk between specific iRef subpopulations and wild type alveolar epithelial cells. In the second aim, gain and loss of function studies of GATA6 will be used to es- tablish a functional link between fibroblast phenotype and alveolarization. In the third aim, H3K27me3 histone marks will be assessed by ChIP-PCR analysis. In vivo and in vitro studies using EZH2 inhibitors will determine a role of EZH2 in suppressing matrix fibroblast signature genes. These studies will provide a better under- standing of the impact of fibroblast diversity on epithelial-mesenchymal crosstalk during alveolarization and alveolar regeneration. This contribution will be significant because it will advance the knowledge about genes and pathways involved in alveolar regeneration, give new insights into the cause of decreased regrowth with age and provide the basis for new therapies to overcome current limitations of regeneration in the human lung. Our understanding of the role of lung fibroblasts in lung injury and regeneration lags far behind our understand- ing of the role of epithelial cells. New information about interstitial lung fibroblasts will be invaluable for defining mechanisms of development, normal lung repair, and lung disease.

项目摘要：虽然在理解上皮亲细胞的作用方面已经取得了巨大进展， 组织修复和纤维化机制中的生殖细胞，但对于其起源、相互作用的性质和基因知之甚少 成纤维细胞祖细胞的调节程序。肺部研究受到了显着的挑战 成纤维细胞群和间充质干细胞群的复杂性和多样性。我们的长期目标 是为了了解成纤维细胞群的复杂性并确定它们对上皮细胞的调节作用 在肺泡化和上皮修复期间。我们的主要目标是确定成纤维细胞分化的关键调节因子 在肺中进行发酵。本申请基于我们将 GATA6 识别为密钥的初步数据 基质 iReF 中的转录因子，对于肺泡化是必不可少的。我们的中心假设是 GATA6 调节 iReF 中的基因表达，从而依次诱导基质 iReF 规范和对位 在发育和修复过程中，crine 与 AEC 相互作用。拟议研究的理由是 了解成纤维细胞亚群的调节及其在上皮间质细胞中的独特作用 相互作用，将为有关肺稳态和修复的基本问题提供新的答案。 该假设将通过三个具体目标进行测试：1）矩阵 iReF 指导 AEC1 分化。 2） GATA6 转录调节一组基质成纤维细胞特征基因，3) H3K27me 沉默 标记调节老化 iReF 中的 GATA6 和基质基因表达。在第一个目标中，肺泡圈培养物将 用于询问特定 iRef 亚群和野生亚群之间的上皮-间质串扰 型肺泡上皮细胞。在第二个目标中，GATA6 功能的获得和丧失研究将用于 es- 建立成纤维细胞表型和肺泡化之间的功能联系。第三个目标，H3K27me3组蛋白 标记将通过 ChIP-PCR 分析进行评估。使用 EZH2 抑制剂的体内和体外研究将确定 EZH2 在抑制基质成纤维细胞特征基因中的作用。这些研究将提供更好的基础 成纤维细胞多样性对肺泡化过程中上皮间质串扰的影响的立场和 肺泡再生。这一贡献将是意义重大的，因为它将推进有关基因的知识 以及参与肺泡再生的途径，为再生减少的原因提供了新的见解 年龄并为新疗法提供基础，以克服目前人肺再生的局限性。 我们对肺成纤维细胞在肺损伤和再生中的作用的理解远远落后于我们的理解—— 上皮细胞的作用。关于间质性肺成纤维细胞的新信息对于定义肺间质成纤维细胞非常有价值 发育机制、正常肺修复和肺部疾病。