Elucidating the FOXF1 gene regulatory network in human alveologenesis

阐明人类肺泡发生中的 FOXF1 基因调控网络

• 批准号: 10558865
• 负责人: Mingxia Gu
• 金额: $ 66.04万
• 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: 2023-02-15 至 2028-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10558865
• 关键词: 3-Dimensional; ATAC-seq; Affect; Alveolar; Alveolar capillary dysplasia with misalignment of pulmonary veins; Benchmarking; Blood Vessels; Blood capillaries; Cell Communication; Cell Nucleus; Cells; Coculture Techniques; Data; Data Set; Deletion Mutation; Development; Disease; Distal; Endoderm; Endothelial Cells; Endothelium; Epithelial Cells; Epithelium; Etiology; Event; FOXF1 gene; Gene Expression; Gene Expression Regulation; Genes; Goals; Heterozygote; Homeostasis; Human; In Vitro; Injury; Lead; Ligands; Lung; Maps; Mediating; Mesoderm; Modeling; Molecular; Mus; Mutation; Organoids; PTEN gene; PTK2 gene; Pathway interactions; Patients; Pattern; Perfusion; Phase; Play; Point Mutation; Population; RNA; Regulation; Role; Sampling; Signal Pathway; Signal Transduction; Site; Structure of parenchyma of lung; System; TGFB2 gene; TGFBR2 gene; Testing; Transforming Growth Factor Beta 2; Transforming Growth Factor beta; Variant; Vascularization; cell type; developmental disease; endothelial stem cell; forkhead protein; gene network; gene regulatory network; genetic signature; genomic locus; improved; induced pluripotent stem cell; injured; insight; lung development; mutant; mutant mouse model; new therapeutic target; novel; postnatal; postnatal human; prenatal; prevent; progenitor; public health relevance; receptor; receptor binding; repaired; single nucleus RNA-sequencing; transcription factor; transcriptomics

PROJECT SUMMARY The overarching goal of the proposed study is to uncover the molecular mechanism underlying FOXF1-mediated temporal and spatial coordination of multiple alveolar endothelial and epithelial cell subtypes. FOXF1 (Forkhead Box F1) is a transcription factor that plays a pivotal role in lung vascular development, homeostasis, and repair after injury. Heterozygous deletions or point mutations in FOXF1 are associated with alveolar-capillary dysplasia with misalignment of pulmonary veins (ACD/MPV), a lethal lung developmental disorder with no cure. Although the Foxf1 mutant mouse model has been created to study ACD, there are certain variations in gene signatures of EC subtypes and developmental timing of the alveolarization in mouse vs. human, limiting its application in understanding ACD human etiology. Thus, it remains unclear how FOXF1 regulates downstream gene networks to drive the formation of alveolar endothelial cell (EC) subtypes and maintain normal EC-epithelial cell (EpiC) crosstalk during alveologenesis in human. Our preliminary studies showed that FOXF1 mutations lead to distinct cell population and transcriptomic changes in two newly identified alveolar EC subtypes- aerocyte (aCap) and general capillary (gCap), based on single nuclei RNA-sequencing in control and ACD human lung tissues. More interestingly, although FOXF1 does not express in epithelial lineage, alveolar type 1 (AT1) EpiC population was significantly reduced, and damage-associated transient epithelial progenitors (DATPs) were increased in FOXF1 mutant human lung. Ligand-receptor binding analysis revealed disrupted TGFβ signaling in aCap (TGFB2)-AT1 (TGFBR2/3) interaction. Therefore, we hypothesized that FOXF1 regulates distinct pathways in aCap and gCap cells, and FOXF1-dependent TGFβ2 secretion by aCap is critical in maintaining AT1 identity and function. We propose to test this hypothesis with the following specific aims: Aim 1: Determine the role of FOXF1 in regulating differentiation and function of alveolar EC subtypes. Aim 2: Determine the impact of aCap cells on maintaining AT1 identity and function. Aim 3: Uncover FOXF1 mediated early lung development using iPSC derived organoid systems. For Aim 3, we established 3D vascularized alveolar organoid models from control and ACD induced pluripotent stem cells (iPSCs), to study FOXF1 gene regulation and dynamic EC-EpiC interactions across multiple lung developmental stages. Completion of the three aims will provide granular mechanistic insights into the spatial and temporal patterning of intercellular signaling pathways driven by FOXF1 in human lung alveologenesis, and provide new therapeutic targets to re-establish normal alveolar-capillary interface in various lung developmental disorders.

项目摘要 这项研究的首要目标是揭示FOXF 1介导的细胞凋亡的分子机制。 多种肺泡内皮和上皮细胞亚型的时间和空间协调。FOXF 1（叉头 Box F1）是一种转录因子，在肺血管发育、稳态和修复中起关键作用 伤后FOXF 1杂合缺失或点突变与肺泡毛细血管发育不良相关 肺静脉错位（ACD/MPV），一种致命的肺部发育障碍，无法治愈。虽然 已经建立了Foxf 1突变小鼠模型来研究ACD，在基因标记中存在某些变异 EC亚型和肺泡化的发育时间在小鼠与人类中的差异，限制了其在 了解ACD的人类病因因此，FOXF 1如何调控下游基因网络仍不清楚 驱动肺泡内皮细胞（EC）亚型的形成，维持正常的EC-上皮细胞（EpiC） 人类肺泡形成过程中的串扰。我们的初步研究表明FOXF 1突变导致不同的 两种新发现的肺泡EC亚型--aCap和aCap的细胞群和转录组学变化 通用毛细管（gCap），基于对照和ACD人肺组织中的单核RNA测序。更 有趣的是，虽然FOXF 1在上皮细胞系中不表达，但肺泡1型（AT 1）EpiC群体在上皮细胞系中表达， 在FOXF 1中，损伤相关的瞬时上皮祖细胞（DATPs）显著减少， 变异人肺配体-受体结合分析显示aCap（TGFB 2）-AT 1中TGFβ信号传导被破坏 （TGFBR 2/3）相互作用。因此，我们假设FOXF 1在aCap和gCap中调节不同的途径， 细胞，并且aCap的FOXF 1依赖性TGFβ2分泌在维持AT 1身份和功能中至关重要。我们 我建议用以下具体目标来检验这一假设：目标1：确定FOXF 1在调节 肺泡EC亚型的分化和功能。目的2：确定aCap细胞对维持 AT 1身份和功能。目的3：使用iPSC衍生的类器官揭示FOXF 1介导的早期肺发育 系统.目的3：建立对照组和ACD诱导组的血管化肺泡类器官模型 多能干细胞（iPSC），研究FOXF 1基因调控和动态EC-EpiC相互作用， 多个肺发育阶段。这三个目标的完成将为以下方面提供精细的机制见解： FOXF 1在人肺中驱动的细胞间信号传导途径的时空模式 肺泡发生，并提供新的治疗目标，以重建正常的肺泡毛细血管界面，在各种 肺发育障碍