Transcriptional clues to esophageal atresia pathogenesis

食管闭锁发病机制的转录线索

基本信息

批准号：
10192781
负责人：
MICHAEL A PACK
金额：
$ 8.13万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10192781
关键词：
Affect Animal Model Architecture Chromosome abnormality Complex Congenital Disorders Data Defect Deglutition Deglutition Disorders Development Disease Distal Dizygotic Twins Eating Embryo Engineering Enteral Esophageal Atresia Esophageal Diseases Esophageal motility disorders Esophagus Esophagus motility Etiology Fishes Functional disorder Funding Ganglia Gastroesophageal reflux disease Gene Expression Gene Expression Regulation Gene Mutation Genes Genetic Genetic Transcription Goals Human Impairment In Vitro Incidence Infant Interstitial Cell of Cajal Intestines Left Length Lesion Live Birth MYH11 gene Malignant neoplasm of esophagus Modeling Morbidity - disease rate Morphology Mus Muscle Muscle Contraction Muscle Development Mutagenesis Mutant Strains Mice Mutation Myosin ATPase Neonatal Nerve Nerve Fibers Nervous System Physiology Nervous system structure Neuroglia Neurons Neurotransmitters Newborn Infant Operative Surgical Procedures Oral Pathogenesis Pathway interactions Patients Phenotype Play Primary Cell Cultures Primitive foregut structure Regulation Regulator Genes Risk Factors Role Siblings Signaling Protein Smooth Muscle Striated Muscles Stromal Cells Supporting Cell Symptoms Syndrome Tissue Grafts Wild Type Mouse Zebrafish base cell motility cell type cohort density differential expression drinking experimental study genetic variant insight macrophage mouse model mutant nerve supply neuromuscular function novel prevent pup response transcriptome sequencing

项目摘要

Project Summary Esophageal atresia (EA) is the most common congenital disorder of the esophagus with a worldwide incidence of about 1 in 3,500 live births. EA can occur as an isolated ﬁnding, or in combination with either syndromic or non-syndromic developmental anomalies. Left untreated, EA is a fatal disorder because the atretic segment obstructs the esophagus. Fortunately, it can be cured surgically in nearly all infants, however the majority suffer from severe gastroesophageal reﬂux and dysphagia as a result of impaired esophageal motility. Although the cause of EA is not known, a genetic component is predicted based on its occurrence in complex developmen- tal syndromes and its higher incidence in monozygotic vs. dizygotic twins. No single gene mutations have been conclusively shown to cause EA. We have developed the ﬁrst animal model of isolated EA by engineering a mutation in the smooth muscle myosin heavy chain gene (Myh11) that we have previously shown alters myosin regulation. In zebraﬁsh, the identical mutation causes invasive expansion of the intestine. In mice the predomi- nant phenotype is EA, although invasive-like intestinal lesions are detected. The mutation disrupts smooth muscle contractility as a result of altered myosin regulation. This suggests that genetic variants in MYH11 and other smooth muscle regulatory genes could cause both EA and its associated esophageal motility disorders. Preliminary transcriptional proﬁling studies of esophagi from Myh11 mutants before the EA phenotype is de- tected showed altered regulation of genes involved in nervous system and striated muscle development. This suggest that defects in esophageal neuromuscular function may be an intrinsic defect associated with EA that arises independently of atresia. Supporting this, immunostainings show altered esophageal nerve ﬁber density, reduced number and size of esophageal ganglia and altered striated muscle development in mutant esophagi The goal of this proposal is to expand these ﬁndings by examining these and other gene expression changes in older Myh11 mutants, by quantifying changes in neuronal density and neuronal subtypes, and by conducting in vitro functional analyses in primary cultures of esophageal neurons.

项目概要食管闭锁（EA）是最常见的食管先天性疾病，在世界范围内都有发病率大约每 3,500 名活产中就有 1 人发生 EA，它可能是一种孤立的发现，也可能与任何一种综合征或其他症状同时发生。如果不治疗，EA 是一种致命的疾病，因为闭锁段。幸运的是，几乎所有婴儿都可以通过手术治愈，但大多数婴儿都患有这种疾病。由于食管蠕动受损而导致严重的胃食管反流和吞咽困难。 EA 的原因尚不清楚，根据其在复杂发育过程中的发生情况可以预测遗传成分 tal 综合征及其在同卵双胞胎与异卵双胞胎中的较高发病率尚未发现单基因突变。最终证明会导致 EA 我们通过设计我们之前已经证明平滑肌肌球蛋白重链基因 (Myh11) 的突变会改变肌球蛋白在斑马鱼中，相同的突变会导致小鼠肠道的侵入性扩张。尽管检测到侵袭性肠道病变，但 Nant 表型为 EA。肌球蛋白调节改变导致肌肉收缩力下降，这表明 MYH11 和 MYH11 的基因变异。其他平滑肌调节基因可能导致 EA 及其相关的食管运动障碍。在 EA 表型脱除之前对 Myh11 突变体食管的初步转录谱研究受保护的结果表明，涉及神经系统和横纹肌发育的基因的调节发生了改变。表明食管神经肌肉功能缺陷可能是与 EA 相关的内在缺陷免疫染色显示食管神经纤维密度改变，支持这一点。突变食管中食管神经节和横纹肌发育的数量和大小减少该提案的目标是通过检查这些和其他基因表达变化来扩展这些发现在较老的 Myh11 突变体中，通过量化神经密度和神经亚型的变化，并通过进行食管神经元原代培养物的体外功能分析。