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

基本信息

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
项目类别：
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

项目摘要

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该领域一名富有成效且有影响力的独立调查员通过进行前沿的多学科研究来更好地了解颅面发育颅面发育和疾病的遗传、分子和细胞机制。我的目标研究的目的是通过阐明疾病机制并促进努力来改善人类健康制定治疗策略。该应用程序将提供通过以下方式实现这些目标的框架：协调的职业发展计划，利用多元化导师和顾问团队的专业知识，以及课程作业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颅面发育和机制的重要问题并将我的训练扩展到新的领域，这对于我向独立的过渡至关重要研究员生涯。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，以及人类颅面解剖学。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转型过程。此外，本次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园区，共同拥有所需的资源、核心设施和教职员工确保本职业发展计划的成功。该应用程序将解决重要但尚未研究的问题 A 型内皮素受体 (EDNRA) 下游细胞内信号转导事件沿着咽弓 1 (PA1) 的背腹 (D-V) 轴的图案域。 EDNRA 信号是下颌发育所必需的，并且在一种称为耳髁的人类颅面疾病中被破坏综合症（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 来模拟 PA1 不同结构域的不同 G 蛋白是未知的，并且极难研究。该提案将使用三个目标来检验 Gq/11 类专门模式化的假设 PA1 的中间结构域，Gq/11 通过诱导来调节模式基因表达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确定 Gq/11 对于中间域图案化是否必要且充分。目标2将使用多模式单细胞分析来定义顺式调控元件和由其控制的基因调控网络 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受指导的博士后转变为准备充分的独立研究员。