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

研究异三聚体 G 蛋白在颅面发育和耳髁综合征中的作用

• 批准号: 10573017
• 负责人: Stanley Michinobu Kanai
• 金额: $ 13.31万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10573017
• 关键词: ATAC-seq; Address; Adult; Affect; Alleles; Anatomy; Area; Automobile Driving; Biology; Branchial arch structure; Career Mobility; Cartilage; Cell Culture Techniques; Cells; Cephalic; ChIP-seq; Chick; Chromatin; Chromatin Remodeling Factor; Colorado; Congenital Abnormality; Connective Tissue; Core Facility; DNA Binding; Data; Defect; Development; Development Plans; Disease; Disease model; Dominant-Negative Mutation; Dorsal; Drug Targeting; Ear; Endothelin Receptor; Endothelin-1; Ensure; Environment; Event; Face; Faculty; First Pharyngeal Arch; Future; G-Protein-Coupled Receptors; GTP-Binding Proteins; Gene Expression; Gene Expression Profile; Genes; Genetic; Goals; Health; Heterotrimeric GTP-Binding Proteins; Human; Immediate-Early Genes; Individual; Interdisciplinary Study; Jaw; Knock-out; Ligands; Link; Mediating; Medical; Mentors; Modeling; Molecular; Molecular Analysis; Morphogenesis; Mus; National Research Service Awards; Neural Crest Cell; Occupations; Pathway interactions; Pattern; Pattern Formation; Phenotype; Phospholipase C; Postdoctoral Fellow; Process; Productivity; Proteins; Publishing; RNA; Receptor Signaling; Regulation; Regulator Genes; Regulatory Element; Reporter; Research; Research Personnel; Research Training; Resources; Role; Scanning; Shapes; Signal Pathway; Signal Transduction; Structure; Surveys; Syndrome; System; Testing; Therapeutic; Training; Transcription Factor AP-1; Transgenic Model; Transgenic Organisms; Translating; Universities; Variant; X-Ray Computed Tomography; XCL1 gene; Zebrafish; bone; career; career development; comparative; craniofacial; craniofacial development; craniofacial disorder; gene network; gene regulatory network; genetic approach; genome-wide; improved; in vitro Assay; in vivo; insight; knockout animal; member; multimodality; novel; pharmacologic; pharyngeal patterning; programs; receptor; recruit; single cell analysis; single cell sequencing; skeletal; success; symposium; training opportunity

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.

我的长期职业目标是成为一名富有成效和有影响力的独立调查员， 颅面发育，通过进行尖端的，多学科的研究，以更好地了解 颅面发育和疾病的遗传、分子和细胞机制。我的目标是 研究的目的是通过阐明疾病机制和促进努力， 制定治疗策略。这个应用程序将提供一个框架， 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。这些因素交织成 我的研究策略解决颅面发育和机制的重要问题， 紊乱，并将我的培训扩展到新的领域，这对我向独立的过渡至关重要。 研究员生涯这包括成为斑马鱼基因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，以及人类颅面解剖学。我 由Clouthier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过渡过程。此外，这项培训将在 颅面生物学系和科罗拉多大学的一流研究环境 安舒茨医学院，共同拥有所需的资源，核心设施和教师， 确保这一职业发展计划的成功。这个应用程序将解决重要的，但未研究 A型内皮素受体（EDNRA）下游的细胞内信号传导事件， 沿咽弓1（PA 1）的背腹（D-V）轴沿着形成图案的区域。EDNRA信号是 这是下颌发育所必需的，在人类颅面疾病耳髁中被破坏 综合征（ARCND）。EDNRA可以通过所有四类G蛋白发出信号，但如何动态使用 通过EDNRA分析不同的G蛋白以使PA 1的不同结构域形成图案是未知的，并且极难研究。 这个提议将使用三个目标来测试Gq/11类专门模式化的假设。 Gq/11通过诱导PA 1中间结构域表达， 全基因组范围的变化，染色质可及性。Aim 1将使用遗传和转基因方法， 确定Gq/11对于中间域图案化是否是必要的和足够的。目标2将使用 多模式单细胞分析，以确定顺式调控元件和基因调控网络控制 Gq/11。目标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，这些信息将用于更好地了解ARCND的人类病例。 总的来说，这个全面的研究培训和职业发展计划将确保我的成功 从一个有指导的博士后过渡到一个准备充分的独立研究员。