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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要拟议研究的总体目标是揭示 FOXF1 介导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。 Box F1) 是一种转录因子，在肺血管发育、稳态和修复中发挥关键作用 FOXF1 杂合缺失或点突变与肺泡毛细血管发育不良有关。肺静脉错位 (ACD/MPV)，这是一种无法治愈的致命性肺部发育障碍。 Foxf1突变小鼠模型是为了研究ACD而创建的，基因特征存在一定的变异小鼠与人类的 EC 亚型和肺泡化的发育时间的差异，限制了其在因此，FOXF1 如何调节下游基因网络仍不清楚。驱动肺泡内皮细胞 (EC) 亚型的形成并维持正常的 EC 上皮细胞 (EpiC) 我们的初步研究表明，FOXF1 突变会导致不同的人类肺泡生成过程中的串扰。两种新发现的肺泡 EC 亚型——有氧细胞 (aCap) 和普通毛细血管 (gCap)，基于对照和 ACD 人肺组织中的单核 RNA 测序。有趣的是，虽然 FOXF1 在上皮谱系中不表达，但肺泡 1 型 (AT1) EpiC 群体 FOXF1 中损伤相关的瞬时上皮祖细胞 (DATP) 减少，而损伤相关的显着瞬时上皮祖细胞 (DATP) 增加突变的人肺配体-受体结合分析显示 aCap (TGFB2)-AT1 中的 TGFβ 信号传导被破坏。 (TGFBR2/3) 相互作用因此，我们阻碍了 FOXF1 调节 aCap 和 gCap 中的不同途径。细胞中，aCap 分泌的 FOXF1 依赖性 TGFβ2 对于维持 AT1 的身份和功能至关重要。建议通过以下具体目标来检验这一假设：目标 1：确定 FOXF1 在调节中的作用目标 2：确定 aCap 细胞对肺泡 EC 亚型的分化和功能的影响。 AT1 的身份和功能：使用 iPSC 衍生的类器官揭示 FOXF1 介导的早期肺发育。对于目标 3，我们根据对照和 ACD 诱导建立了 3D 血管化肺泡类器官模型。多能干细胞 (iPSC)，用于研究 FOXF1 基因调控和动态 EC-EpiC 相互作用多个肺部发育阶段的完成将为我们提供详细的机制见解。人肺中 FOXF1 驱动的细胞间信号通路的时空模式肺泡发生，并提供新的治疗靶点，以在各种情况下重建正常的肺泡-毛细血管界面肺部发育障碍。