Role of alveolar fibroblasts in extracellular matrix organization and alveolar type 1 cell differentiation

肺泡成纤维细胞在细胞外基质组织和肺泡1型细胞分化中的作用

• 批准号: 10731854
• 负责人: Anne-Karina Theresia Perl
• 金额: $ 80.01万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10731854
• 关键词: Address; Adhesions; Adult; Alveolar; Alveolus; Animal Model; Basal Cell; Basal lamina; Binding; Biological Assay; Bronchopulmonary Dysplasia; Cell Communication; Cell Differentiation process; Cell Proliferation; Cells; ChIP-seq; Characteristics; Chromatin; Chronic lung disease; Collagen; Data; Data Set; Deposition; Development; Dysplasia; EZH2 gene; Engraftment; Epigenetic Process; Epithelial Cells; Epithelium; Extracellular Matrix; Fibroblasts; Fibrosis; Future; GATA6 transcription factor; Gene Expression; Genes; Genetic; Genetic Transcription; Genomics; Goals; Homeostasis; Human; Hyperoxia; Hyperplasia; Impairment; In Vitro; Infant; Knowledge; Lipids; Lung; Lung diseases; Membrane; Mission; Modeling; Modification; Molecular; Mus; Natural regeneration; Nature; Nuclear; Organoids; Paracrine Communication; Patients; Perinatal; Population; Premature Infant; Proteomics; Public Health; Pulmonary Emphysema; Regulation; Regulatory Element; Replacement Therapy; Reporting; Repression; Reproducibility; Research; Role; Signal Transduction; Site; Specific qualifier value; Structure; Testing; Tissues; Transcriptional Regulation; United States National Institutes of Health; WNT Signaling Pathway; alveolar epithelium; cofactor; comparative; experimental study; fibroblast growth factor 18; fibrotic lung disease; gain of function; idiopathic pulmonary fibrosis; in vitro Model; induced pluripotent stem cell; inhibitor; innovation; insight; intercellular communication; loss of function; lung injury; lung regeneration; overexpression; paracrine; perinatal period; permissiveness; pharmacologic; postnatal; promoter; regenerative therapy; repaired; restoration; self-renewal; small hairpin RNA; therapy development; tool; transcription factor; transcriptomics; transdifferentiation

ABSTRACT: An in-depth understanding of the mechanisms that regulate alveolar septation and septal wall maturation will be required to develop therapies for premature infants with Bronchopulmonary Dysplasia (BPD) and for developing potential therapies for adult lung regeneration. We and others have begun to define changes in PDGFRa myo, lipo and matrix fibroblast (FB) function during alveolarization, homeostasis, regeneration, and fibrosis. Our long-term goal is elucidation of molecular regulators of FB functions and how diverse FBs support epithelial cells. Our objective herein is to identify molecular mech- anisms that result in functional changes in alveolar fibroblasts that regulate ECM organization and epithelial differentia- tion. Integration of transcriptomic datasets predicted GATA6 as a regulator of PDGFRa-FB differentiation. Our preliminary data show that conditional inactivation of GATA6 in perinatal PDGFRa+ FBs resulted in loss of matrix and gain in lipo fibro- blast function comparable to findings in BPD and animal models of hyperoxia. GATA6 inactivation in PDGFRa+ FBs also resulted in fragmented collagen and detachment of AT1 cells from the basal lamina demonstrating a thus far unidentified role of alveolar FBs in ECM organization. Inactivation of GATA6 resulted in significant increase of the lipo FB transcription factor TCF21 expression and preliminary in vitro data suggest suppression of TCF21 by GATA6. The central hypothesis is that GATA6 and transcriptional cofactors regulate FB function in alveolar FBs by suppressing lipo FB differentiation. The rationale for this research is a new understanding of the regulation of matrix FB function and how these functionally different stages modify extracellular matrix and the FB-epithelial crosstalk. Aim 1 will test the hypothesis that GATA6 suppresses lipo FB function and promotes ECM deposition in the alveolar FBs. This aim will use lineage tracing to deter- mine trans-differentiation of the alveolar FB to a lipo FB. And use in vitro cell derived matrices to identify ECM modifica- tions by lipo and matrix FBs. Aim 2 will test the hypothesis that GATA6 is in a transcriptional network that regulates lipo and matrix FB function. In this aim we will use in vitro gain and loss of function of GATA6 and TCF21 and identify and validate transcriptional networks and partnering transcription factors. Aim 3 will test the hypothesis that GATA6 regulates ECM synthesis and paracrine signaling in PDGFRa+ FBs that direct AT2 to AT1 differentiation. The proposed studies in this application will identify the transcriptional regulation of matrix FB function and their role in extracellular matrix deposition and cell-cell signaling in the alveolar niche. The proposed research is conceptually innovative, because we ask questions regarding the nature of fibroblast plasticity and functional switches. The proposed research is scientifically innovative, because the contractile function of alveolar fibroblasts, has been thoroughly investigated, but little is known about their matrix organizing function or their role in supporting AT2 proliferation and AT1 differentiation This contribution will be significant because it addresses 1) lack of knowledge of matrix and signaling function of the alveolar FB; 2) identifies and validates the transcriptional network around GATA6 that regulates FB function.

抽象的： 需要深入了解调节肺泡间隔和间隔壁成熟的机制 为患有支气管肺发育不良（BPD）的早产儿开发治疗方法并开发潜在的治疗方法 用于成人肺再生。我们和其他人已经开始定义 PDGFRa 肌、脂肪和基质成纤维细胞 (FB) 的变化 肺泡化过程中的功能， 稳态、再生和纤维化。我们的长期目标是阐明分子 FB 功能的调节因子以及不同的 FB 如何支持上皮细胞。我们的目标是确定分子机制 导致肺泡成纤维细胞功能改变的现象，调节 ECM 组织和上皮分化 。转录组数据集的整合预测 GATA6 作为 PDGFRa-FB 分化的调节剂。我们的初步 数据显示，围产期 PDGFRa+ FB 中 GATA6 的条件失活导致基质损失和脂肪纤维增加。 原始细胞功能与 BPD 和高氧动物模型中的发现相当。 PDGFRa+ 中 GATA6 失活 FB还有 导致胶原碎片和 AT1 细胞从基底层脱离，这表明迄今为止尚未鉴定出 肺泡 FB 在 ECM 组织中的作用。 GATA6 失活导致 lipo FB 转录显着增加 TCF21 因子表达和初步体外数据表明 GATA6 抑制 TCF21。中心假设是 GATA6 和转录辅助因子通过抑制脂肪 FB 分化来调节肺泡 FB 中的 FB 功能。这 这项研究的基本原理是对基质 FB 功能的调节以及这些功能如何发挥作用有了新的认识 不同阶段会改变细胞外基质和 FB-上皮串扰。目标 1 将检验 GATA6 的假设 抑制脂质 FB 功能并促进 ECM 在肺泡 FB 中沉积。这一目标将使用谱系追踪来阻止 我的肺泡FB转分化为脂FB。并使用体外细胞衍生基质来鉴定 ECM 修饰- 脂质和基质 FB 的作用。目标 2 将检验 GATA6 位于调节脂质的转录网络中的假设 和矩阵FB函数。为此，我们将利用 GATA6 和 TCF21 的体外功能获得和丧失，并识别和 验证转录网络和合作转录因子。目标 3 将检验 GATA6 调节的假设 PDGFRa+ FB 中的 ECM 合成和旁分泌信号传导指导 AT2 向 AT1 分化。本研究中提出的研究 应用将确定基质 FB 功能的转录调控及其在细胞外基质沉积中的作用 以及肺泡微环境中的细胞间信号传导。拟议的研究在概念上具有创新性，因为我们提出问题 关于成纤维细胞可塑性和功能开关的性质。所提出的研究具有科学创新性， 因为肺泡成纤维细胞的收缩功能已被彻底研究，但对其的了解甚少 矩阵组织函数或 它们在支持 AT2 增殖和 AT1 分化中的作用这一贡献将是 意义重大，因为它解决了 1) 缺乏对肺泡 FB 矩阵和信号功能的了解； 2) 识别和 验证围绕 GATA6 调节 FB 功能的转录网络。