Identifying Gli-Regulated Targets during Tracheal Development and Disease

识别气管发育和疾病过程中 Gli 调节的靶点

• 批准号: 10195097
• 负责人: Talia S Nasr
• 金额: $ 4.57万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-20 至 2022-04-19
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10195097
• 关键词: Address; Bioinformatics; Biological Process; Cadherins; Cartilage; Cell Adhesion; Cells; Cellular biology; Childhood; Chondrogenesis; Clinical Treatment; Data; Data Set; Defect; Development; Diagnosis; Disease; Embryo; Epithelial; Epithelium; Equilibrium; Erinaceidae; Esophagus; Etiology; Exhibits; Extracellular Matrix; FOXF1 gene; Failure; Fetal Development; GLI3 gene; Gene Expression Profiling; Genes; Genetic; Genetic Predisposition to Disease; Genomics; Goals; Human; Impairment; In Situ; In Situ Hybridization; In Vitro; Lead; Learning; Lesion; Literature; Mediating; Mesenchymal; Mesenchymal Differentiation; Mesenchyme; Metalloproteases; Modeling; Molecular; Morphogenesis; Mus; Mutation; Online Mendelian Inheritance In Man; Pallister-Hall syndrome; Pathway interactions; Patients; Perinatal; Physicians; Prenatal Diagnosis; Primitive foregut structure; Process; Proteins; Publishing; Research; Review Literature; Scientist; Series; Signal Transduction; Smooth Muscle; Smooth Muscle Myocytes; Testing; Therapeutic; Tissues; Trachea; Tracheal Atresia; Tracheoesophageal Fistula; Training; Transgenic Organisms; Tube; WNT Signaling Pathway; Xenopus; base; cartilage development; causal variant; clinical Diagnosis; data mining; exome sequencing; experimental study; fetal; human tissue; induced pluripotent stem cell; loss of function; mouse genetics; mouse model; mutant; novel therapeutics; overexpression; programs; respiratory; smoothened signaling pathway; transcription factor; transcriptome sequencing; treatment strategy; virtual

Project Summary/Abstract This goal of this project is to identify the mechanisms by which Hedgehog/Gli signaling regulates tracheal development and how disruptions in this process can lead to perinatal tracheal defects. Errors in tracheal development may range from tracheal atresia and tracheoesophageal (TE) clefts, where the fetal foregut tube fails to separate properly, to tracheomalacia due to impaired tracheal chondrogenesis. The etiology of tracheal defects is poorly understood and new diagnoses and therapies are needed to address these relatively common yet potentially lethal defects. Human patient exome sequencing as well as murine mutant models have begun to identify a few developmental pathways that regulate tracheal development, including the Hedgehog/Gli pathway. For example, in murine models a total loss of Gli2 and Gli3 results in a complete failure of tracheal development, while some compound heterozygous Gli2;Gli3 mutants exhibit a TE cleft with tracheomalacia. In addition, our preliminary data suggests that transgenic expression of a Gli3 mutant that mimics the genetic lesion found in human Pallister-Hall syndrome patients also leads to impaired TE separation and virtually absent tracheal cartilage. The specific aims of this project are to identify Gli target genes in the developing trachea, to determine how Gli regulates TE separation and tracheal cartilage differentiation, and to model the mechanism of human tracheal defects. Preliminary RNA-seq studies of control and mutant foreguts have begun to define Gli targets during TE development. I will test the hypothesis that these candidate Gli target genes are misexpressed and/or mislocalized during TE development in Gli mutants. Our preliminary further data suggest that Wnt and BMP pathways act downstream of HH/Gli, and we will use a combination of mouse genetics and fetal foregut cultures to test the hypothesis that defective Wnt and BMP signaling is responsible for the defects in tracheal chondrogenesis. The results of these experiments will inform the basis of tracheal defects in human patients and as well as guide differentiation of tracheal iPSCs towards new therapeutic possibilities. This project will also provide valuable training in current clinical diagnosis and treatment strategies for these patients. !

项目摘要/摘要 这个项目的目标是确定Hedgehog/Gli信号调节气管的机制 以及这一过程中的中断如何导致围产期气管缺陷。气管中的错误 发育的范围可能包括气管闭锁和气管食道(TE)裂，在这些地方胎儿的前肠管 不能正确分离，导致气管软化，因为气管软骨生成受损。气管的病因学 人们对缺陷知之甚少，需要新的诊断和治疗方法来解决这些相对常见的问题 然而，潜在的致命缺陷。人类患者外显子组测序和小鼠突变模型已经开始 确定一些调节气管发育的发育途径，包括Hedgehog/Gli 路径。例如，在小鼠模型中，Gli2和Gli3的完全缺失会导致气管完全衰竭 一些复合杂合型Gli2；Gli3突变体表现为TE裂伴气管软化。在……里面 此外，我们的初步数据表明，模拟基因的Gli3突变体的转基因表达 在人类Pallister-Hall综合征患者中发现的损害也会导致TE分离受损和 气管软骨缺失。 这个项目的具体目标是在发育中的气管中识别Gli靶基因，以确定Gli如何 调节TE的分离和气管软骨的分化，并模拟人类气管的机制 缺陷。对对照和突变前景的初步rna-seq研究已经开始确定Gli靶标 TE开发。我将测试这些候选Gli靶基因被错误表达和/或 Gli突变体在TE发育过程中定位错误。我们的初步进一步数据表明，WNT和 BMP途径在HH/Gli下游发挥作用，我们将结合使用小鼠遗传学和胎儿前瞻 培养以验证WNT和BMP信号缺陷导致气管缺陷的假说 软骨生成。这些实验的结果将为人类患者的气管缺陷提供依据。 以及引导气管IPSCs分化为新的治疗可能性。这个项目将 还为这些患者提供当前临床诊断和治疗策略方面的有价值的培训。 好了！