Molecular aspects of CRKL in heart development and human disease

CRKL 在心脏发育和人类疾病中的分子方面

基本信息

批准号：
9197022
负责人：
BERNICE E MORROW
金额：
$ 81万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE, INC
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-15 至 2020-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9197022
关键词：
22q11 Deletion Syndrome 22q11.2 Accounting Adaptor Signaling Protein Affect Anterior Aortic Valve Stenosis Back Base Pairing Biological Assay Boxing CRKL gene Candidate Disease Gene Cardiac Cell physiology Cells Cessation of life Chromosomes Clinical Code Complex Congenital Heart Defects DNA Data Defect Development DiGeorge Syndrome Disease Embryo Endocardium Etiology Evaluation Focal Adhesion Kinase 1 Genes Genetic Genetic Predisposition to Disease Genetic Transcription Heart Heart Abnormalities Heart Valves Human Human Genetics In Vitro Individual Intracardiac abnormalities Investigation MAP Kinase Gene Mediating Mesenchymal Molecular Mus Mutant Strains Mice Mutation Neural Crest Cell Outcome Pathway interactions Patients Persistent Truncus Arteriosus Pharyngeal Apparatus Phenotype Population Process Rare Diseases Receptor Protein-Tyrosine Kinases Regulatory Element Reporting Research Risk Sampling Septate Shprintzen syndrome Signal Pathway Signal Transduction System Testing Tetralogy of Fallot Tissues Untranslated RNA Validation Variant Vascular Endothelial Growth Factors Work base cardiogenesis cohort congenital heart disorder conotruncal anomaly face syndrome differential expression disease heterogeneity embryo culture exome sequencing gene function gene interaction genetic approach genome sequencing human disease in vitro Assay in vivo insight loss of function malformation middle age mouse model mutant novel paracrine proband programs transcription factor transcriptome sequencing whole genome

22q11 Deletion Syndrome 22q11.2 Accounting Adaptor Signaling Protein Affect Anterior Aortic Valve Stenosis Back Base Pairing Biological Assay Boxing CRKL gene Candidate Disease Gene Cardiac Cell physiology Cells Cessation of life Chromosomes Clinical Code Complex Congenital Heart Defects DNA Data Defect Development DiGeorge Syndrome Disease Embryo Endocardium Etiology Evaluation Focal Adhesion Kinase 1 Genes Genetic Genetic Predisposition to Disease Genetic Transcription Heart Heart Abnormalities Heart Valves Human Human Genetics In Vitro Individual Intracardiac abnormalities Investigation MAP Kinase Gene Mediating Mesenchymal Molecular Mus Mutant Strains Mice Mutation Neural Crest Cell Outcome Pathway interactions Patients Persistent Truncus Arteriosus Pharyngeal Apparatus Phenotype Population Process Rare Diseases Receptor Protein-Tyrosine Kinases Regulatory Element Reporting Research Risk Sampling Septate Shprintzen syndrome Signal Pathway Signal Transduction System Testing Tetralogy of Fallot Tissues Untranslated RNA Validation Variant Vascular Endothelial Growth Factors Work base cardiogenesis cohort congenital heart disorder conotruncal anomaly face syndrome differential expression disease heterogeneity embryo culture exome sequencing gene function gene interaction genetic approach genome sequencing human disease in vitro Assay in vivo insight loss of function malformation middle age mouse model mutant novel paracrine proband programs transcription factor transcriptome sequencing whole genome

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项目摘要

ABSTRACT One of the greatest challenges in human genetics is to understand the genetic basis of complex disorders, such as congenital heart disease (CHD). Studies of rare diseases with known genetic etiologies such as the 22q11.2 deletion syndrome (DiGeorge/velo-cardio-facial syndrome), can pinpoint disease genes and mechanisms. Approximately 65% of 22q11DS patients have a congenital heart defect, mostly affecting the cardiac outflow tract (OFT). These range from mild to severe and include persistent truncus arteriosus and tetralogy of Fallot, at the severe end. Although TBX1, encoding a T-box transcription factor has been the strongest candidate gene for CHD on chromosome 22q11.2, patients with similar heart defects, but non- overlapping deletions within this region have been identified. In these patients with atypical deletions, CRKL, encoding a cytoplasmic adaptor to receptor tyrosine kinase signaling is deleted, but not TBX1. This implicates CRKL as a true disease gene. Although there have been extensive studies of TBX1, CRKL is understudied with respect to heart development. The neural crest cells are a critical cell population in the pharyngeal apparatus needed for proper alignment of the cardiac OFT. We propose to explore neural crest cell functions of Crkl by evaluating new upstream and downstream pathways in Aim 1. We will then delineate the subset that is critical for OFT development as mediated by Tbx1 and Crkl. Crkl-/- mice die at E16.5, earlier than can be expected based upon only having OFT defects. Our pathological investigation has found that Crkl-/- embryos have OFT valve defects in addition to alignment malformations. OFT valve anomalies are highly relevant to 22q11DS, as 9% of patients have these defects. This is a newly recognized defect for which the molecular mechanisms are not known. Based upon this, and the presence of similar defects in endocardial-specific null mutant embryos, we hypothesize that Crkl is required in this tissue to form derivative OFT valves via VEGF- MAPK and additional signaling pathways. Endocardial functions will be tested in Aim 2. This work is also clinically important because middle-aged 22q11DS patients have now been reported to have sudden unexplained death, unconnected to the well-known intracardiac malformations. This might be related to the presence of aortic valve stenosis, which accounts for 6.6% of 22q11.2 deletion patients. To connect the mouse work in Aims 1 and 2 back to the human patients, we will evaluate existing whole exome sequence (WES) from the current largest cohort that we are aware, of >1,000, 22q11DS patients for mutations in CRKL interaction network genes in Aim 3. Exploratory studies of non-coding sequences in the candidate genes will be performed. Validation will be done using more DNA samples from 500 22q11DS subjects and from emerging WES data on non-syndromic CHD patients. The synergy of mouse and human studies will offer rapid discoveries for a deep understanding of the complex genetic etiology of CHD.

抽象的人类遗传学中最大的挑战之一是了解复杂疾病的遗传基础，例如先天性心脏病（CHD）。研究罕见疾病具有已知遗传病因，例如 22Q11.2缺失综合征（Digeorge/Velo-Cardio-Facial综合征），可以查明疾病基因和机制。大约22q11ds患者中约有65％患有先天性心脏缺陷，主要影响心脏流出区（通常）。这些范围从轻度到重度，包括持续的动脉truncus anteriosus和 Fallot的四部曲，处于严重的末端。尽管TBX1，编码T-box转录因子已经是染色体22q11.2的CHD的最强候选基因，心脏缺陷相似但非 - 已经确定了该区域内的重叠缺失。在这些非典型删除患者中，CRKL，删除了对受体酪氨酸激酶信号传导的细胞质适配器的编码，但未删除TBX1。这意味着 CRKL作为真正的疾病基因。尽管对TBX1进行了广泛的研究，但CRKL已研究关于心脏发展。神经rest细胞是咽部的关键细胞种群适当对齐心脏OFT所需的设备。我们建议探索神经rest细胞功能通过评估AIM 1中的新上游和下游途径的CRKL。然后，我们将描绘该子集对于由TBX1和CRKL介导的FAST开发至关重要。 crkl - / - 小鼠死于E16.5，早于预期仅基于经常缺陷。我们的病理研究发现CRKL - / - 胚胎除了对齐畸形外，还有经常存在阀门缺陷。经常阀异常与 22q11ds，因为9％的患者患有这些缺陷。这是一个新认识的缺陷机制尚不清楚。基于此，以及内膜特异性null中的相似缺陷突变的胚胎，我们假设该组织中需要CRKL才能通过VEGF-形成衍生物OFT阀 MAPK和其他信号通路。心内膜功能将在AIM 2中进行测试。这项工作也是临床上很重要，因为现在据报道中年22q11ds患者无法解释的死亡，与众所周知的心脏内畸形无关。这可能与主动脉瓣狭窄的存在，占22q11.2缺失患者的6.6％。连接鼠标在目标1和2回到人类患者中，我们将评估现有的整个外显子组序列（WES）我们知道的当前最大的队列，> 1,000，22q11ds患者用于CRKL相互作用的突变 AIM 3中的网络基因。候选基因中非编码序列的探索性研究将是执行。将使用来自500 22q11ds受试者的更多DNA样品和新兴的DNA样品进行验证 WES数据上的非综合症冠心病患者。老鼠和人类研究的协同作用将迅速深入了解CHD复杂遗传病因的发现。