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且有影响力的独立调查员 颅面发展，通过进行尖端的多学科研究，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 研究是通过阐明疾病机制来改善人类健康，并为努力做出贡献 制定治疗策略。该应用程序将提供一个框架，以实现这些目标 协调的职业发展计划，利用多元化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。这些因素被编织成 我的研究策略解决了颅面发展和机制中的重要问题 疾病并将我的培训扩展到我向独立过渡至关重要的新领域 研究人员职业。这包括成为斑马鱼基因组的遗传和转基因模型的专家 诸如单细胞ATAC-SEQ和CHIP-SEQ以及人类颅面解剖结构等广泛的测序方法。我的 导师团队，由博士领导。 Clouthier和Nichols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，该培训将在 颅面生物学系和科罗拉多大学的出色研究环境 Anschutz Medical Campus，共同拥有资源，核心设施和教职员工需要 确保该职业发展计划的成功。该应用程序将解决重要但未研究 内皮素受体A型（EDNRA）下游的细胞内信号传导中的事件（EDNRA） 咽弓1（PA1）沿背腹侧（D-V）轴的图案域。 Ednra信号是 下颌发育的必需品，并在称为Auriculocondylar的人类颅面疾病中被破坏 综合征（ARCND）。 Ednra可以通过所有四类G蛋白发出信号，尽管如何动态使用 埃德拉（Ednra）与PA1的不同域的不同G蛋白是未知的，并且非常难以研究。 该提案将使用三个目标来检验以下假设：GQ/11类专门模板 PA1的中间结构域，该GQ/11通过诱导调节基因表达 全基因组对染色质的可及性的变化。 AIM 1将使用遗传和转基因方法 确定GQ/11是否需要且足以用于中间域模式。 AIM 2将使用 多模式的单细胞分析以定义由控制的顺式调节元件和基因调节网络控制 GQ/11。 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 信号。此外，这些信息将用于更好地了解人类的ARCND病例。 总的来说，这项全面的研究培训和职业发展计划将确保我的成功 从指导的博士后过渡到准备充分的独立研究人员。