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.

项目摘要 先天性肺部疾病是严重的疾病，过早地结束了许多人的生命， 在他们的一生中幸存的个人的负担。先天性膈疝（CDH）就是其中之一。 导致肺严重发育不全的疾病。虽然疝气可以 手术修复后，肺缺损是持久的，导致高死亡率和复杂的长期护理， 幸存者有许多遗传因素导致CDH，其中一些基因改变了肺部 发展独立于疝气的影响，显示其对正常的内在重要性。 肺的生长。通过对CDH患者的基因组和转录组学分析的综合， 确定了两个基因，KMT 2D和KDM 6A作为介导严重肺发育不全的强有力的候选者。KMT2D （赖氨酸甲基转移酶2D）和KDM 6A（赖氨酸脱甲基酶6A）是合作的表观遗传调节因子 其在CDH中可以突变和下调，并且已知对干细胞很重要。新 技术进步使我们能够从气管吸出物中分离肺上皮基底干细胞 CDH患者和对照组患者。我们将利用这些细胞来测试这些表观遗传功能的丧失是否 调节因子影响这些细胞自我更新和产生分化的肺上皮的能力， 找出它们控制的下游基因。我们的具体目标是：1）确定病原体的影响 KMT 2D/KDM 6A遗传变异和药物干预对患者分化/自我更新的影响 衍生自气管吸出物的衍生的肺基底干细胞，和2）映射H3 K4 me 1和H3 K27 me 3 在KMT 2D/KDM 6A突变或抑制后CDH和对照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， 可能与一整类肺部疾病有关。这些目标将帮助我们了解 控制肺发育的机制出错，使我们能够开发治疗方法来纠正这些问题。 问题