Investigating the Role of Heterotrimeric G Proteins in Craniofacial Development and Auriculocondylar Syndrome

我的长期职业目标是成为一名富有成效和影响力的独立调查员 通过进行前沿、多学科的研究，更好地了解 颅面发育和疾病的遗传、分子和细胞机制。我的目标是 研究是通过阐明疾病机制并为以下努力做出贡献来改善人类健康 制定治疗策略。此应用程序将提供框架，以通过 协调的职业发展计划，利用不同的导师和顾问团队的专业知识，以及 课程和会议提供的补充培训机会。这些因素交织在一起 我的研究策略解决了颅面发育中的重要问题和机制 混乱并将我的培训扩展到新的领域，这些领域对于我过渡到独立的 研究人员的职业。这包括成为斑马鱼遗传和转基因模型的专家，基因组- 广泛的测序方法，如单细胞ATAC-SEQ和CHIP-SEQ，以及人类颅面解剖。我的 由Clouthier博士和Nichols博士领导的导师团队将在整个培训计划中提供宝贵的指导， 包括学术求职和职业转型的过程。此外，这项培训将在IN进行。 科罗拉多大学颅面生物学系一流的研究环境 Anschutz医学院，它们共同拥有所需的资源、核心设施和教职员工 确保这一职业发展计划的成功。本申请将解决尚未研究的重要问题 内皮素受体A型(Ednra)下游的细胞内信号转导事件 沿咽弓1(PA1)背腹(D-V)轴的图案域。Ednra信令是 是下颌骨发育所必需的，在一种名为耳髁的人类头面部疾病中被破坏 综合征(ARCND)。Ednra可以通过所有四类G蛋白发出信号，尽管如何动态地使用 不同的G蛋白通过Ednra形成PA1的不同结构域是未知的，也是非常困难的研究。 这项提议将使用三个目标来测试GQ/11类独占地 PA1中间结构域，GQ/11通过诱导 染色质可及性的全基因组变化。目标1将使用遗传和转基因方法来 确定GQ/11对于中间域图案化是否必要且充分。AIM 2将使用 多模式单细胞分析确定顺式调控元件和基因调控网络 AIM 3将使用ARCND的斑马鱼模型来了解疾病等位基因如何影响Ednra-GQ/11 发信号。此外，这些信息将被用来更好地了解人类ARCND病例。 总体而言，这项全面的研究、培训和职业发展计划将确保我成功 从有指导的博士后过渡到准备充分的独立研究人员。

My long-term career goal is to become a productive and impactful independent investigator in the field of craniofacial development, by conducting cutting-edge, multidisciplinary research to better understand the genetic, molecular, and cellular mechanisms of craniofacial development and disorders. The goal of my research is to improve human health by elucidating disease mechanisms and contributing towards efforts to develop therapeutic strategies. This application will provide the framework to achieve these goals through a coordinated career development plan that utilizes the expertise of a diverse team of mentors and advisors, and the supplemental training opportunities provided by coursework and conferences. These factors are woven into my research strategy addressing important questions in craniofacial development and mechanisms of disorders and expands my training into new areas that are essential for my transition to an independent researcher career. This includes becoming an expert in genetic and transgenic models of zebrafish, genome- wide sequencing approaches like single cell ATAC-seq and ChIP-seq, and human craniofacial anatomy. My mentor team, led by Drs. Clouthier and Nichols, will provide valuable guidance throughout the training plan, including the academic job search and career transition process. Moreover, this training will be carried out at in a stellar research environment in the Department of Craniofacial Biology, and the University of Colorado Anschutz Medical Campus, that collectively have the resources, core facilities, and faculty members needed to ensure the success of this career development plan. This application will address important yet unstudied events in intracellular signaling downstream of the Endothelin receptor type A (EDNRA) that establishes patterning domains along the dorsal-ventral (D-V) axis of pharyngeal arch 1 (PA1). EDNRA signaling is required for lower jaw development and is disrupted in a human craniofacial disorder called Auriculocondylar syndrome (ARCND). EDNRA can signal through all four classes of G proteins, though how dynamic use of different G proteins by EDNRA to pattern different domains of PA1 is unknown and extremely difficult to study. This proposal will use three aims to test the hypothesis that the Gq/11 class exclusively patterns the intermediate domain of PA1, and that Gq/11 regulates patterning gene expression by inducing genome-wide changes to chromatin accessibility. Aim 1 will use genetic and transgenic approaches to determine whether Gq/11 is necessary and sufficient for intermediate domain patterning. Aim 2 will use multimodal single-cell analysis to define cis-regulatory elements and gene regulatory networks controlled by Gq/11. Aim 3 will use zebrafish models of ARCND to understand how disease alleles impact EDNRA-Gq/11 signaling. Furthermore, this information will be used to better understand human cases of ARCND. Collectively, this comprehensive research training and career development plan will ensure my successful transition from a mentored postdoc to a well-prepared independent researcher.