Identifying the role of dynamic ECM-derived forces in zebrafish semicircular canal morphogenesis

识别动态 ECM 衍生力在斑马鱼半规管形态发生中的作用

基本信息

批准号：
10547914
负责人：
Akankshi Munjal
金额：
$ 24.9万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-06-01 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10547914
关键词：
Ablation Acceleration Address Alagille Syndrome Atlases Award Biological Models Biophysics Biosensor CHARGE syndrome CRISPR/Cas technology Cells Cellular biology Collaborations Complex Congenital Abnormality Data Set Deposition Development Developmental Biology Disease Ear Diseases Embryo Epithelial Equilibrium Etiology Event Extracellular Matrix Feedback Fingers Generations Genes Genetic Genetic Transcription Glycosaminoglycans Goals Grant Growth HAS3 gene Head Heparan Sulfate Proteoglycan Heparitin Sulfate Hyaluronan Hydrogels Image Image Analysis Interdisciplinary Study Jaw Laboratories Labyrinth Lasers Liquid substance Literature Maps Mechanics Mediating Mentors Mentorship Microfluidics Modeling Molecular Morphogenesis Organ Otic Vesicle Pathway interactions Pattern Phase Physics Polymers Postdoctoral Fellow Process Production Property Proteoglycan Publications Publishing Quantitative Microscopy Regenerative Medicine Regulation Reporter Research Resolution Role Semicircular canal structure Sequence Analysis Shapes Signal Transduction Simple Epithelium Specific qualifier value Swelling Syndrome Technology Testing Theoretical model Tissues Training Transgenic Organisms Translational Research Velocimetries Vertebrates Water Work Writing Xenopus Zebrafish automated image analysis base biophysical model biophysical properties biophysical tools career career development experience fundamental research gene synthesis high resolution imaging imaging genetics improved in vivo inner ear diseases insight mechanical force mechanical properties meetings morphogens mutant next generation novel particle pressure programs quantitative imaging response single cell sequencing single-cell RNA sequencing skills student mentoring tool transcription factor transcriptome versican zebrafish development

项目摘要

PROJECT SUMMARY The form of an organ defines its function. How simple epithelial tissues give rise to the complex forms of mature organs is a fundamental question in developmental biology with important implications in disease and regenerative medicine. A beautiful example is the remodeling of a simple embryonic epithelium to form the three semicircular canals (SCC) of the inner ear. Their intricate form is required for sensing balance and acceleration and is conserved across all jawed vertebrates. SCCs can develop abnormally in congenital syndromes such as LAMM, CHARGE and Alagille syndrome, for unknown reasons. The mechanisms underlying healthy development are poorly understood. My preliminary findings indicate that local synthesis of a hyaluronan-rich extracellular matrix (ECM) is important in SCC development. In the proposed research, I will determine how this ECM helps generate mechanical forces in SCC morphogenesis in zebrafish (Aim1). I will use next-generation single-cell sequencing to identify genes involved in the patterning of an active ECM (Aim2) and to identify and characterize the role of other ECM components involved in SCC development (Aim3). The novel molecular- mechanical principles revealed from this study will be widely applicable to other morphogenetic events involving an active ECM and topological epithelial remodeling. Moreover, as these processes are disrupted in many congenital diseases and ear disorders, my findings will improve our understanding of these diseases. The proposed research draws on 1) my training in quantitative cell biology in a development context, 2) the new skills acquired in my post-doctoral research, including zebrafish research and imaging of the developing organ at single-cell resolution, and 3) my K99 training plan to implement automated image analysis, in vivo biophysical measurements, modeling, and single-cell sequencing and analysis. To fulfil my aims, I will receive mentorship from Drs. Sean Megason, Tim Mitchison, L. Mahadevan and Allon Klein, whose combined expertise in zebrafish development, gene-editing, biophysics, quantitative imaging, modeling and single-cell RNA sequencing will give me the training I need to commence my independent research program. To facilitate my career development during the mentored K99 phase, I will take relevant courses and maintain collaborations for both fundamental and translational research, present my work at meetings, publish my research, write grants, and gain further experience in management and mentoring students to formulate my own mentoring style. I will greatly benefit from the experience, track record and regular feedback from my co-mentors to start my own laboratory. My long-term career goal is to head an inter-disciplinary research program to investigate the mechanical and molecular basis of morphogenesis and to use the insights I develop to advance translational research. I’ve made significant progress towards this goal in the form of research experience, successful collaborations and publications. The K99 award will give me access to the additional training and expertise I need to successfully transition to independence.

项目摘要器官的形式定义了其功能。简单的上皮组织产生了成熟的复杂形式器官是发育生物学的基本问题，对疾病和再生医学。一个美丽的例子是简单的胚胎上皮的重塑以形成三个内耳的半圆形管（SCC）。它们的复杂形式是灵敏度平衡和加速度所必需的并在所有颌脊椎动物中保守。 SCC可以在先天性综合征中发展绝对出于未知原因，Lamm，Charge和Alagille综合征。健康的基础机制发展知之甚少。我的初步发现表明，透明质酸富含透明质酸的局部合成细胞外基质（ECM）在SCC开发中很重要。在拟议的研究中，我将确定如何 ECM有助于在斑马鱼中的SCC形态发生中产生机械力（AIM1）。我将使用下一代单细胞测序以识别参与活性ECM（AIM2）模式的基因，并识别和识别和表征与SCC开发有关的其他ECM组件的作用（AIM3）。新颖的分子 - 这项研究揭示的机械原理将广泛适用于其他涉及的形态发生事件活跃的ECM和拓扑上皮重塑。而且，由于这些过程在许多过程中被破坏先天性疾病和耳部疾病，我的发现将改善我们对这些疾病的理解。拟议的研究借鉴了1）我在开发环境中对定量细胞生物学的培训，2）新的在我的博士后研究中获得的技能，包括斑马鱼研究和发展器官的成像在单细胞分辨率和3）我的K99培训计划以实施自动图像分析，体内生物物理测量，建模和单细胞测序和分析。为了实现我的目标，我将获得Mentalship 来自博士。 Sean Megason，Tim Mitchison，L。Mahadevan和Allon Klein，其在斑马鱼中的专业知识开发，基因编辑，生物物理学，定量成像，建模和单细胞RNA测序将提供我需要培训我的独立研究计划。促进我的职业发展在指导的K99阶段，我将参加相关课程并维持这两种基本的合作并翻译研究，介绍我在会议上的工作，发布我的研究，写赠款并进一步获得具有管理和心理学生的经验，可以制定我自己的心理风格。我会很大的好处根据经验，记录和我的委托人的定期反馈，以开始自己的实验室。我的长期职业目标是领导一项跨学科研究计划，以调查机械和形态发生的分子基础，并利用我发展的见解来推进翻译研究。我做了以研究经验，成功合作和出版物。 K99奖将使我获得成功的额外培训和专业知识过渡到独立。