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 拟议的研究的总体目标是发现FOXF1介导的分子机制 多个肺泡内皮和上皮细胞亚型的临时和空间配位。 FOXF1（叉子） Box F1）是转录因子，在肺血管发育，体内平衡和维修中起关键作用 受伤后。 FOXF1中的杂合缺失或点突变与肺泡毛细血管发育不良有关 随着肺静脉（ACD/MPV）的未对准，一种致命的肺发育障碍，无法治愈。虽然 FOXF1突变小鼠模型是为了研究ACD的创建，基因特征存在某些变化 EC亚型和在小鼠中发展肺泡化的时机，并限制了其在 了解ACD人类病因。那尚不清楚FOXF1如何调节下游基因网络 驱动肺泡内皮细胞（EC）亚型的形成并保持正常的ec上皮细胞（EPIC） 人类肺单元期间的串扰。我们的初步研究表明，FOXF1突变导致不同 两种新鉴定的肺泡EC亚型的细胞种群和转录组变化（ACAP）和 一般毛细管（GCAP），基于对照和ACD人类肺组织中的单核RNA测序。更多的 有趣的是，尽管FOXF1在上皮谱系中没有表达，但肺泡1（AT1）的史诗人口是 在FOXF1中，显着降低和与损害相关的瞬态上皮祖细胞（DATP）增加了 突变的人肺。配体受体结合分析显示，ACAP中TGFβ信号传导破坏（TGFB2）-at1 （TGFBR2/3）相互作用。因此，我们假设FOXF1调节ACAP和GCAP中的不同途径 ACAP通过ACAP分泌细胞，而FOXF1依赖性TGFβ2分泌对于维持AT1身份和功能至关重要。我们 提出以以下特定目的检验该假设的建议：目标1：确定FOXF1在调节中的作用 肺泡EC亚型的分化和功能。目标2：确定ACAP细胞对维护的影响 AT1身份和功能。 AIM 3：使用IPSC衍生的Organoid探索FOXF1介导的早期肺发育 系统。对于AIM 3，我们从对照和ACD诱导的 多能干细胞（IPSC）研究FOXF1基因调节和跨动态ECEPIC相互作用 多个肺发育阶段。这三个目标的完成将为颗粒状的机械洞察力提供 由FOXF1在人肺中驱动的细胞间信号通路的空间和临时模式 肺单元术，并提供新的治疗靶标，以重新建立各种正常的肺泡毛细血管界面 肺发育障碍。