Discovery and pathogenic characterization of novel monogenic causes of bladder dysfunction.

项目总结 标题：膀胱功能障碍新的单基因病因的发现和致病特征 先天性肾脏和尿路异常(CAKUT)是慢性肾脏病最常见的原因 儿童时期的肾脏疾病。已确定40多种CAKUT的单基因病因，另有150种 基因已被描述为导致尿路畸形是一个整体的综合征 组件。然而，这些基因中的大多数结合成了调节上尿路的途径。 发育，如后肾间充质转化和输尿管芽形成。其发病机制及发病机制 下尿路疾病的遗传调控仍然知之甚少。最近，通过使用完整的 外显子组测序，我发现烟碱型乙酰胆碱受体α3-亚基突变为 继发性CAKUT引起膀胱功能障碍的新原因。这是仅有的8个已知单基因之一 先天性膀胱功能障碍的原因，独特的是，是一种调节自主神经系统的基因 功能。我假设可以确定下尿路疾病的其他单基因原因。 通过使用整个外显子组测序，另外，这些基因将结合在一起 调节尿路上皮信号、逼尿肌收缩和膀胱神经的通路。 拟议研究的目的有两个：(1)同时应用候选基因和无偏分析 利用外显子组测序数据寻找膀胱功能障碍新的单基因病因的策略 以及(2)使用完整外显子组和完整转录组测序的综合基因组方法 一个患有CAKUT的国际儿童队列。上述拟议研究的完成将提供 对控制膀胱发育和功能的病理机制的进一步洞察，并最终， 可能为膀胱功能障碍儿童带来新的治疗策略。此外，验证 尿转录组测序方法有助于克服目前在整个外显子组中的局限性 剪接改变基因变异的鉴定和解释中的测序。 这个K08应用程序包含一个全面的职业发展计划，不仅确保进步 并成功地进行了拟议的研究，但也促进了我向独立的 调查员。它包括与我的导师和咨询委员会、研究和职业生涯的定期会议 发展研讨会、基因组学和转录组学课程，以及参加国家和 国际会议。这项研究将在波士顿丰富的学术环境中进行 儿童医院和哈佛医学院为进一步培训和培训提供了理想的环境 职业发展。

PROJECT SUMMARY Title: Discovery and pathogenic characterization of novel monogenic causes of bladder dysfunction Congenital anomalies of the kidney and urinary tract (CAKUT) are the most common cause of chronic kidney disease in childhood. Over 40 monogenic etiologies of CAKUT have been identified, and another 150 genes have been described to cause syndromes in which urinary tract malformations are an integral component. However, the majority of these genes coalesce into pathways that regulate upper urinary tract development, such as metanephric mesenchyme transition and ureteric bud formation. The pathogenesis and genetic regulation of lower urinary tract disease remain poorly understood. Recently, through the use of whole exome sequencing, I identified mutations in the α3-subunit of the nicotinic acetylcholine receptor, CHRNA3, as a novel cause of bladder dysfunction with secondary CAKUT. This is one of just eight known monogenic causes of congenital bladder dysfunction, and, uniquely, is a gene that regulates autonomic nervous system function. I hypothesize that additional monogenic causes of lower urinary tract disease can be identified through the use of whole exome sequencing, and additionally, that these genes will coalesce upon pathways that regulate urothelial signaling, detrusor contraction, and bladder innervation. The aims of the proposed research are twofold: (1) to apply both a candidate gene and unbiased analysis strategy to whole exome sequencing data in order to discover novel monogenic causes of bladder dysfunction and (2) to apply an integrated genomic approach using whole exome and whole transcriptome sequencing to an international cohort of children with CAKUT. Accomplishment of the proposed research above will provide additional insight into the pathomechanisms that govern bladder development and function, and, ultimately, could lead to novel therapeutic strategies for children with bladder dysfunction. In addition, validation of a method for urinary transcriptome sequencing can help to overcome current limitations in whole exome sequencing in the identification and interpretation of splice-altering genetic variants. This K08 application encompasses a comprehensive career development plan to not only ensure progress and success in carrying out the proposed research, but also to facilitate my transition to an independent investigator. It encompasses regular meetings with my mentor and advisory committee, research and career development seminars, courses in genomics and transcriptomics, and participation at national and international conferences. The research will be conducted in the rich academic environment at Boston Children's Hospital and Harvard Medical School, which provides an ideal environment for further training and career development.