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.

项目摘要 致死性肺发育障碍(LLDD)很少被诊断出来，但具有破坏性的肺发育不良。 (PHS)，表现为进行性新生儿缺氧和严重的肺动脉高压(PAH)。基座 在组织病理学上，LLDDS传统上被归类为肺泡毛细血管异常增殖症 肺静脉错位(ACDMPV)、腺泡发育不良(AcDys)、先天性肺泡发育不良(CAD)、 和其他未指明的主要小灵通。我们发现杂合性单核苷酸变异(SNV)在 间质转录因子FOXF1基因或拷贝数变异(CNV)缺失涉及FOXF1 或其位于上游约300kb的肺特异性增强子在80-90%的患者中对ACDMPV起作用。我们 报道称，该增强子还上调了FOXF1附近的顺式lncRNA Fendrr图谱。有趣的是， 与SNV不同，CNV的缺失几乎完全发生在母亲的16号染色体上。最近，我们和其他人 证明了另一种间充质转铁蛋白、TBX4和旁分泌信号中的变异所起的作用 在患有AcDys、CAD和其他原发PHS的婴儿中，超过60%的婴儿存在FGF10分子，表明 Tbx4-FGF10信号在LLDDS发病机制中的意义。重要的是，FOXF1和TBX4的变种 也与更常见的特发性或家族性儿童PAH有关。有趣的是，我们发现了一个 慢性粒细胞白血病患者FOXF1和TBX4增强子中非编码SNV的显著丰富 LLDD的可变表示形式。此外，我们的CHIP-SEQ和RNA-SEQ研究暗示了 SHH-FOXF1和TBX4-FGF10信号通路，涉及鲜为人知的肺内皮细胞 跨膜蛋白TMEM100。我们假设(I)监管区域内的非编码SNV 肺发育基因可以显著改变(减轻或加重)LLDD和PAH的表型，(Ii)和 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LLDD和PAH家族中，以及(Iii)SHH-FOXF1和TBX4-FGF10信号通路的相互作用， 涉及TMEM100，是人类肺正常发育所必需的。使用人类肺标本和细胞系 和小鼠模型，我们将识别和分析FOXF1在慢性阻塞性肺疾病患者中的非编码调控元件。 ACDMPV和/或PAH(Aim 1)以及Tbx4和FGF10在AcDys、CAD、其他PHS和/或 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- 间充质信号，涉及TMEM100，以解开复杂的化合物遗传 LLDDS和PAH。我们的研究将阐明人类肺发育的遗传学以及它的扰动如何 转化为低密度脂蛋白和多环芳烃的表型变异。我们将识别新的基因变异，允许更多 对这些疾病的准确诊断和预后，促进更多信息丰富的遗传咨询，以及 为开发LLDDS和PAH的宫内治疗潜力提供目标。我们的数据还将有助于 更好地理解人类遗传学中的不完全外显和表达能力变化现象。