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分离和成熟肺泡壁的机制 开发出支气管肺发育不良（BPD）和开发潜在疗法的过早婴儿的疗法 用于成人肺部再生。我们和其他人已经开始定义PDGFRA MYO，LIPO和基质成纤维细胞（FB）的变化 牙槽化期间的功能， 稳态，再生和纤维化。我们的长期目标是阐明分子 FB功能的调节剂以及不同的FBS如何支持上皮细胞。我们的目的是确定分子机械 导致肺泡成纤维细胞的功能变化的anism，从而调节ECM组织和上皮差异 tion。转录组数据集的集成预测GATA6是PDGFRA-FB分化的调节剂。我们的初步 数据表明，围产期PDGFRA+ FBS中GATA6的条件失活导致基质损失和Lipo纤维中的增益 爆炸功能可与BPD和多氧动物模型中的发现相当。 pDGFRA+中的GATA6失活 FBS也是 导致碎屑碎片胶原蛋白和AT1细胞从基底层脱离，表现出迄今为止未识别的 肺泡FB在ECM组织中的作用。 GATA6的灭活导致Lipo Fb转录显着增加 TCF21因子表达和初步的体外数据表明GATA6抑制了TCF21。中心假设是 该GATA6和转录辅助因子通过抑制脂肪FB分化来调节肺泡FB的FB功能。这 这项研究的理由是对矩阵FB函数调节的新理解以及它们在功能上如何 不同阶段修改细胞外基质和FB上皮串扰。 AIM 1将测试GATA6的假设 抑制Lipo FB功能并促进肺泡FBS中的ECM沉积。这个目标将使用谱系跟踪来阻止 地雷将牙槽FB射向Lipo Fb。并使用体外细胞衍生的矩阵识别ECM修饰 - Lipo和基质FBS的tions。 AIM 2将测试GATA6在调节Lipo的转录网络中的假设 和矩阵FB功能。在此目标中，我们将使用GATA6和TCF21的体外增益和功能丧失，并识别和 验证转录网络和合作转录因子。 AIM 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将AT2直接定位为AT1分化。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 和肺泡小裂中的细胞细胞信号传导。拟议的研究在概念上是创新的，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。拟议的研究在科学上是创新的， 由于肺泡成纤维细胞的收缩功能已经进行了彻底的研究，但对它们的收缩功能知之甚少 矩阵组织功能或 它们在支持AT2增殖和AT1分化中的作用这一贡献将是 重要的是因为它解决了1）缺乏肺泡Fb的基质和信号传导功能的知识； 2）识别和 验证调节FB功能的GATA6周围的转录网络。