Etiology and pathogenesis of lethal lung developmental disorders in neonates

新生儿致命性肺发育障碍的病因和发病机制

• 批准号: 10660107
• 负责人: PAWEL STANKIEWICZ
• 金额: $ 79.34万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-15 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10660107
• 关键词: Alleles; Alveolar; Alveolar capillary dysplasia with misalignment of pulmonary veins; Appearance; Autopsy; Binding Sites; Biological Assay; Biopsy; Cell Line; ChIP-seq; Childhood; Chromosome 16; Classification; Code; Complex; Congenital alveolar dysplasia; Copy Number Polymorphism; DNA Binding; Data; Development; Developmental Gene; Diagnosis; Disease; Distant; Dysplasia; Electrophoretic Mobility Shift Assay; Embryo; Endothelium; Enhancers; Epithelium; Etiology; FGF10 gene; FGFR2 gene; FOXF1 gene; Family; Genes; Genetic; Genetic Counseling; Heart Abnormalities; Heterozygote; Histologic; Histones; Human; Human Cell Line; Human Genetics; Infant; Integral Membrane Protein; Life; Lung; Maps; Mediating; Mediator; Mesenchymal; Mesenchyme; Modification; Mus; Mutate; Nucleic Acid Regulatory Sequences; Paracrine Communication; Pathogenesis; Pathogenicity; Patients; Penetrance; Phase; Phenotype; Play; Process; Prognosis; Prognostic Marker; Promoter Regions; RNA Interference; Regulatory Element; Reporting; Research; Resolution; Role; SHH gene; Signal Pathway; Signal Transduction; Signaling Molecule; Single Nucleotide Polymorphism; Specimen; Techniques; Technology; Translating; Untranslated RNA; Variant; developmental disease; diagnostic biomarker; disease phenotype; fetal; genetic variant; genome editing; hypoxia neonatorum; in utero; insight; interest; lung development; malformation; mouse model; neonate; novel; novel diagnostics; prevent; promoter; pulmonary arterial hypertension; pulmonary hypoplasia; therapeutic target; transcription factor; transcriptome sequencing

Project Summary Lethal lung developmental disorders (LLDDs) are rarely diagnosed but devastating pulmonary hypoplasias (PHs), presenting with progressive neonatal hypoxia and severe pulmonary arterial hypertension (PAH). Based on histopathological appearance, LLDDs have been traditionally classified as alveolar capillary dysplasia with misalignment of pulmonary veins (ACDMPV), acinar dysplasia (AcDys), congenital alveolar dysplasia (CAD), and other unspecified primary PHs. We found that heterozygous single nucleotide variants (SNVs) in the mesenchymal transcription factor (TF) FOXF1 gene or copy-number variant (CNV) deletions involving FOXF1 or its lung-specific enhancer located ~ 300 kb upstream are responsible for ACDMPV in 80-90% of patients. We reported that this enhancer also up-regulates in cis lncRNA FENDRR mapping nearby FOXF1. Interestingly, unlike SNVs, CNV deletions arise almost exclusively on the maternal chromosome 16. Recently, we and others demonstrated the causative role for variants in another mesenchymal TF, TBX4, and a paracrine signaling molecule FGF10 in greater than 60% of infants with AcDys, CAD, and other primary PHs, indicating the significance also of TBX4-FGF10 signaling in pathogenesis of LLDDs. Importantly, FOXF1 and TBX4 variants have been associated also with more common idiopathic or familial childhood PAH. Interestingly, we found a statistically significant enrichment of non-coding SNVs in the FOXF1 and TBX4 enhancers in patients with variable presentation of LLDD. Moreover, our ChIP-seq and RNA-seq studies have implied interactions between the SHH-FOXF1 and TBX4-FGF10 signaling pathways, involving little-known lung-specific endothelial transmembrane protein TMEM100. We hypothesize that (i) non-coding SNVs within the regulatory regions of lung developmental genes can dramatically modify (alleviate or exacerbate) LLDD and PAH phenotypes, (ii) an interplay between the coding and non-coding variants can explain the complex compound inheritance observed in families with LLDDs and PAH, and (iii) interaction of SHH-FOXF1 and TBX4-FGF10 signaling pathways, involving TMEM100, is required for proper human lung development. Using human lung specimens and cell lines and mouse models, we will identify and analyze non-coding regulatory elements of FOXF1 in patients with ACDMPV and/or PAH (Aim 1) and those of TBX4 and FGF10 in patients with AcDys, CAD, other PHs, and/or PAH (Aim 2). In Aim 3, we will decipher the crosstalk between SHH-FOXF1 and TBX4-FGF10 epithelial- mesenchymal signaling, involving TMEM100, to untangle the complex compound inheritance in families with LLDDs and PAH. Our studies will elucidate the genetics of lung development in humans and how its perturbations translate to phenotypic variability of LLDDs and PAH. We will identify new genetic variants, allowing for more precise diagnosis and prognosis of these disorders, facilitating more informative genetic counseling, and providing targets for development of potential in utero treatments for LLDDs and PAH. Our data will also help to better understand incomplete penetrance and variable expressivity phenomena in human genetics in general.

项目概要 致命性肺发育障碍 (LLDD) 很少被诊断出来，但却是毁灭性的肺发育不全 （PH），表现为进行性新生儿缺氧和严重肺动脉高压（PAH）。基于 根据组织病理学表现，LLDD 传统上被归类为肺泡毛细血管发育不良 肺静脉错位 (ACDMPV)、腺泡发育不良 (AcDys)、先天性肺泡发育不良 (CAD)、 和其他未指定的主要 PH。我们发现杂合单核苷酸变异（SNV） 间充质转录因子 (TF) FOXF1 基因或涉及 FOXF1 的拷贝数变异 (CNV) 缺失 或其位于上游约 300 kb 的肺特异性增强子负责 80-90% 患者的 ACDMPV。我们 据报道，该增强子还在 FOXF1 附近的顺式 lncRNA FENDRR 中上调。有趣的是， 与 SNV 不同，CNV 缺失几乎完全出现在母体 16 号染色体上。最近，我们和其他人 证明了另一种间充质 TF TBX4 和旁分泌信号传导中的变异的致病作用 超过 60% 患有 AcDys、CAD 和其他原发性 PH 的婴儿体内存在 FGF10 分子，这表明 TBX4-FGF10 信号传导在 LLDD 发病机制中也具有重要意义。重要的是，FOXF1 和 TBX4 变体 也与更常见的特发性或家族性儿童 PAH 相关。有趣的是，我们发现了一个 患有以下疾病的患者中 FOXF1 和 TBX4 增强子中非编码 SNV 的富集具有统计学意义 LLDD 的变量呈现。此外，我们的 ChIP-seq 和 RNA-seq 研究暗示了两者之间的相互作用 SHH-FOXF1 和 TBX4-FGF10 信号通路，涉及鲜为人知的肺特异性内皮细胞 跨膜蛋白TMEM100。我们假设 (i) 监管区域内的非编码 SNV 肺发育基因可以显着改变（减轻或加剧）LLDD 和 PAH 表型，(ii) 编码和非编码变体之间的相互作用可以解释观察到的复杂复合遗传 在患有 LLDD 和 PAH 的家族中，以及 (iii) SHH-FOXF1 和 TBX4-FGF10 信号通路的相互作用， 涉及 TMEM100，是人类肺部正常发育所必需的。使用人肺标本和细胞系 和小鼠模型，我们将鉴定并分析患有以下疾病的患者中 FOXF1 的非编码调控元件： ACDMPV 和/或 PAH（目标 1）以及 AcDys、CAD、其他 PH 和/或患者中的 TBX4 和 FGF10 PAH（目标 2）。在目标 3 中，我们将破译 SHH-FOXF1 和 TBX4-FGF10 上皮细胞之间的串扰 涉及 TMEM100 的间充质信号传导，以解开家族中复杂的复合遗传 LLDD 和 PAH。我们的研究将阐明人类肺部发育的遗传学及其扰动 转化为 LLDD 和 PAH 的表型变异。我们将识别新的基因变异，从而允许更多 这些疾病的精确诊断和预后，促进提供更多信息的遗传咨询，以及 为 LLDD 和 PAH 子宫内治疗潜力的开发提供目标。我们的数据也将有助于 更好地理解人类遗传学中的不完全外显率和可变表达现象。