Understanding the impact of targeting the epigenetic regulation of lung stem cells in congenital lung disease

了解针对肺干细胞表观遗传调控对先天性肺病的影响

• 批准号: 10726411
• 负责人: Samuel Philip Rowbotham
• 金额: $ 16.7万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 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 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10726411
• 关键词: Affect; Basal Cell; Cause of Death; Cells; Complex; Congenital Abnormality; Congenital diaphragmatic hernia; DNA Sequence Alteration; Defect; Development; Disease; Engineering; Epigenetic Process; Epithelium; Genes; Genetic; Genetic Transcription; Genomics; Growth; Health; Hernia; Individual; Intervention; Life; Long-Term Care; Lung; Lung diseases; Lysine; Maps; Measures; Mediating; Methyltransferase; Mutate; Mutation; Newborn Infant; Operative Surgical Procedures; Organ; Pathogenicity; Patients; Pharmacologic Substance; Program Development; Regulator Genes; Structure; Survivors; Testing; Trachea; Variant; aspirate; cohort; design; developmental disease; epigenetic regulation; genetic variant; histone demethylase; histone methyltransferase; inhibitor; loss of function; lung development; mortality; new technology; premature; progenitor; recruit; repaired; self-renewal; stem cell self renewal; stem cells; therapy development; transcriptomics

Project Summary Congenital lung diseases are serious disorders which prematurely end the lives of many and place large health burdens on surviving individuals throughout their lives. Congenital Diaphragmatic Hernia (CDH) is one such disease which causes severe underdevelopment (hypoplasia) of the lung. Although the hernia can be surgically repaired, the lung defects are persistent, leading to high mortality and complicated long-term care for survivors. There are many genetic factors contributing to CDH, and some of these genes change lung development independently from the effects of the hernia, showing their intrinsic importance to the normal growth of the lung. Through a synthesis of genomic and transcriptomic analyses of CDH patients we have identified two genes, KMT2D and KDM6A as strong candidates to mediate the severe lung hypoplasia. KMT2D (Lysine Methyltransferase 2D) and KDM6A (Lysine Demethylase 6A) are co-operating epigenetic regulators which can be both mutated and downregulated in CDH and are known to be important to stem cells. New technological advances have allowed us to isolate lung epithelial basal stem cells from the tracheal aspirates of recruited CDH and control patients. We will utilize these cells to test if the loss of function of these epigenetic regulators affects the ability of these cells to self-renew and generate differentiated lung epithelium, and to identify the downstream genes which they control. Our specific aims are 1) Determine the effects of pathogenic KMT2D/KDM6A genetic variants and pharmaceutical interventions on the differentiation/self-renewal of patient- derived lung basal stem cells derived from tracheal aspirates, and 2) Map the H3K4me1 and H3K27me3 epigenetic changes in CDH and control basal stem cells following mutation or inhibition of KMT2D/KDM6A. With this approach we will be taking advantage of the genetic breakthroughs in CDH to make discoveries that are potentially relevant to a whole class of lung diseases. These aims will help us understand how the mechanisms governing lung development go awry and enable us to develop therapies to correct these problems.

项目概要 先天性肺病是一种严重的疾病，它会过早地结束许多人的生命并损害人们的健康 幸存者一生的负担。先天性膈疝（CDH）就是其中之一 导致肺部严重发育不良（发育不全）的疾病。虽然疝气可以 经过手术修复后，肺部缺陷持续存在，导致高死亡率和复杂的长期护理 幸存者。导致 CDH 的遗传因素有很多，其中一些基因会改变肺部 发育独立于疝气的影响，显示出其对正常生命的内在重要性 肺部的生长。通过对 CDH 患者的基因组和转录组分析进行综合分析，我们发现 确定了两个基因 KMT2D 和 KDM6A 作为介导严重肺发育不全的有力候选基因。 KMT2D （赖氨酸甲基转移酶 2D）和 KDM6A（赖氨酸脱甲基酶 6A）是协同作用的表观遗传调节因子 它可以在 CDH 中突变和下调，并且已知对干细胞很重要。新的 技术进步使我们能够从气管吸出物中分离出肺上皮基底干细胞 招募的 CDH 患者和对照患者。我们将利用这些细胞来测试这些表观遗传功能是否丧失 调节因子影响这些细胞自我更新和生成分化肺上皮的能力，并 识别它们控制的下游基因。我们的具体目标是 1) 确定致病因素的影响 KMT2D/KDM6A 基因变异和药物干预对患者分化/自我更新的影响 源自气管抽吸物的肺基底干细胞，以及 2) 绘制 H3K4me1 和 H3K27me3 的图谱 KMT2D/KDM6A 突变或抑制后 CDH 和控制基底干细胞的表观遗传变化。 通过这种方法，我们将利用 CDH 的遗传突破来做出以下发现： 可能与一整类肺部疾病相关。这些目标将帮助我们了解如何 控制肺部发育的机制出错，使我们能够开发出纠正这些机制的疗法 问题。