Prdm3/16 Regulate Chromatin Accessibility to Determine Alveolar Maturation

Prdm3/16 调节染色质可及性以确定肺泡成熟度

基本信息

批准号：
10636860
负责人：
DANIEL, T (MD) Todd Swarr
金额：
$ 70.5万
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：
2022
资助国家：
美国
起止时间：
2022-07-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10636860
关键词：
ATAC-seq Acute Lung Injury Acute Respiratory Distress Syndrome Adult Alveolar Alveolus Architecture Binding Biochemical Biological Assay Birth Bronchopulmonary Dysplasia Cell Differentiation process Cell Lineage Cell Maturation Cell physiology Cells ChIP-seq Chromatin Chronic lung disease Co-Immunoprecipitations Complex Complex Mixtures Data Development Developmental Biology Disease EVI1 gene Embryo Endothelium Enhancers Epigenetic Process Epithelial Cells Epithelium Family Fetal Lung Fingers Foundations Future Gases Gene Expression Gene Expression Regulation Genes Genetic Materials Genetic Transcription Genome Genomics Health Histones Homeostasis Impairment In Vitro Inflammation Influenza A virus Injury Label Laboratories Life Lipids Liquid Chromatography Luciferases Lung Lung diseases Mediating Molecular Morphogenesis Mus Natural regeneration Neonatal Respiratory Distress Organoids Pathogenesis Perinatal Phospholipids Physiological Pregnancy Premature Infant Preparation Production Proliferating Proline-Rich Domain Promoter Regions Proteins Proteomics Pulmonary Surfactants Pulmonary alveolar structure Regulation Regulatory Element Reporter Respiratory Failure Role Signal Transduction Site Specific qualifier value Structure Surface Surface Tension Testing Transcript Transgenic Mice Viral Zinc Fingers airway epithelium alveolar epithelium alveolar homeostasis alveolar lamellar body cell type cofactor developmental genetics epigenome fetal gene regulatory network genomic locus histone methyltransferase improved in vitro Assay in vivo injury and repair lung development lung injury lung maturation lung repair member postnatal prenatal prevent progenitor protein expression protein protein interaction pulmonary function recruit repaired respiratory distress syndrome stem cells surfactant surfactant deficiency tandem mass spectrometry transcription factor transcriptome sequencing

项目摘要

PROJECT SUMMARY PRDMs regulate chromatin accessibility to determine alveolar maturation. Pulmonary surfactant is a complex mixture of lipids and proteins produced by AT2 (Alveolar Type 2 Cells) which is secreted into alveolar spaces to reduce surface tension and prevent alveolar collapse during the ventilatory cycle. Lack of pulmonary surfactant underlies the pathogenesis of neonatal respiratory distress syndrome (RDS) in preterm infants and contributes to the pathogenesis of acute lung injury (ARDS) in adults. The fetal lung undergoes dramatic changes in architecture, cell differentiation and gene expression in preparation for birth, increasing surfactant lipid and protein production required for postnatal lung function. While our laboratory and others have identified genes, transcriptional regulators, and gene regulatory networks controlling surfactant homeostasis, how chromatin accessibility is modulated to enable gene transcription necessary for AT1 and AT2 cell maturation and function remains relatively unexplained. Lacking are data regarding the molecular mechanisms controlling chromatin accessibility required for the activation of critical transcription factors and their targets before birth and during alveolar repair. The present application is based on our recent identification of the primary roles of PRDM3 and PRDM16 in regulation of genes critical for alveolar maturation, remodeling, and gene expression required for postnatal survival. PRDM3/16 are zinc finger, proline-rich domain-containing transcription factors with histone modifying activity regulating chromatin accessibility throughout the genome. We have shown that deletion of Prdm3/16 in fetal respiratory epithelial cells caused respiratory failure at birth, dramatically impairing surfactant protein production. We will use ATAC-Seq, Cut and Run, and RNA-Seq to identify gene loci regulated by PRDM3/16 to infer mechanisms by which PRDM proteins interact with AT2 specific genes before birth. In this application, we will test the hypothesis that PRDM3 and PRDM16 interact in transcriptional complexes regulating recruitment of histones and chromatin accessibility controlling AT2 cell differentiation and function in late gestation and during regeneration following viral-induced lung injury in adult mice. We will identify and test the functions of binding partners and transcriptional complexes mediating PRDM3/16 activity. The gene regulatory networks, physiologic and biochemical roles of PRDM3/16 on AT2 cells, alveolar structure and function will be identified in vivo and in vitro. The proposed studies represent conceptual advances regarding the molecular control of alveolar epithelial cell differentiation and function and will provide a framework for the development of future strategies to modify epigenetic landscapes controlling AT2/AT1 cell function in health and disease. Rev: 9-3-21

项目摘要 PRDMS调节染色质的可及性，以确定肺泡成熟。肺表面活性剂是由AT2（肺泡2型细胞）产生的脂质和蛋白质的复合混合物，该混合物分泌成牙槽降低表面张力并防止肺泡塌陷的空间。缺乏肺部表面活性剂是早产儿（RDS）新生儿呼吸窘迫综合征（RDS）的发病机理的基础有助于成人急性肺损伤（ARDS）的发病机理。胎儿肺发生了巨大变化在体系结构中，细胞分化和基因表达在准备出生时，表面活性剂脂质增加和产后肺功能所需的蛋白质产生。尽管我们的实验室和其他人都确定了基因，但转录调节剂和控制表面活性剂稳态的基因调节网络如何染色质可访问性调节以启用AT1和AT2细胞成熟所需的基因转录和功能仍然相对无法解释。缺乏有关控制染色质的分子机制的数据激活关键转录因子及其出生之前和期间的目标所需的可及性牙槽修复。本申请基于我们最近对PRDM3和 PRDM16在调节基因对肺泡成熟，重塑和基因表达至关重要的基因中所需的基因表达至关重要产后生存。 PRDM3/16是锌指，富含组蛋白的含有脯氨酸的转录因子修改整个基因组中调节染色质可及性的活性。我们已经证明了删除的胎儿呼吸性上皮细胞中的PRDM3/16引起出生时呼吸衰竭，急剧损害表面活性剂蛋白质产生。我们将使用atac-seq，剪切和运行，以及RNA-seq来识别受其调节的基因基因座 PRDM3/16推断PRDM蛋白在出生前与AT2特定基因相互作用的机制。在这个应用，我们将测试PRDM3和PRDM16在转录复合物中相互作用的假设调节组蛋白和染色质可及性的募集，以控制AT2细胞分化和病毒诱导成年小鼠肺损伤后妊娠和再生期间的功能。我们将识别和测试结合伴侣和转录复合物的功能，介导PRDM3/16 活动。 PRDM3/16在AT2细胞上的基因调节网络，生理和生化作用，肺泡结构和功能将在体内和体外鉴定。拟议的研究代表概念进步关于肺泡上皮细胞分化和功能的分子控制，并将提供一个框架为了制定未来的策略，以修改控制AT2/AT1细胞功能的表观遗传景观健康与疾病。 Rev：9-3-21