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：
9978320
负责人：
MICHAEL A PACK
金额：
$ 8.1万
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 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9978320
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.

项目摘要食道闭锁（EA）是食道最常见的先天性疾病，全球发病率大约有3,500名活产中的1个。 EA可以作为孤立的发现或与综合症或结合非综合发育异常。剩下的未治疗，ea是一种致命的疾病阻塞食道。幸运的是，几乎所有婴儿都可以通过手术治愈，但是多数患者由于食管运动受损而导致严重的胃食管反流和吞咽困难。虽然 EA的原因尚不清楚，遗传成分是根据其在复杂发育中的出现而预测的 TAL综合征及其在单卵双胞胎与双卵双胞胎中的发病率更高。没有单个基因突变是最终证明会导致EA。我们通过工程A开发了第一个孤立EA的动物模型我们以前显示的平滑肌肌球蛋白重链基因（MYH11）中的突变改变了肌球蛋白规定。在斑马鱼中，相同的突变会导致肠的浸润性膨胀。在小鼠中 NANT表型是EA，尽管检测到了浸润性的肠道病变。突变破坏了光滑肌球蛋白调节的改变导致肌肉收缩力。这表明MYH11和其他平滑肌调节基因可能会引起EA及其相关的食管运动障碍。在EA表型之前，来自MyH11突变体的食管的初步转录研究研究指示显示了与神经系统和肌肉发育相关的基因的调节改变。这表明食管神经肌肉功能的缺陷可能是与EA相关的固有缺陷独立于闭锁。支持这一点，免疫染色显示食管神经纤维密度改变，食管神经节的数量和大小减少，并改变了突变食管突变的肌肉发育该提案的目的是通过检查这些基因表达变化来扩展这些发现在较旧的MyH11突变体中，通过量化神经元密度和神经元亚型的变化，并通过进行食管神经元原发性培养物中的体外功能分析。