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：
10393115
负责人：
Akankshi Munjal
金额：
$ 5.11万
依托单位：
HARVARD MEDICAL SCHOOL
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 2021-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10393115
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 Structure 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.

项目概要器官的形式决定了其功能。简单的上皮组织如何产生成熟的复杂形式。器官是发育生物学中的一个基本问题，对疾病和疾病具有重要影响。再生医学的一个很好的例子是重塑一个简单的胚胎上皮以形成这三个细胞。内耳的半规管（SCC）结构复杂，是感知平衡和加速度所必需的。并且在所有有颌脊椎动物中都是保守的，鳞状细胞癌可以在先天性综合症中异常发育，例如 LAMM、CHARGE 和 Alagille 综合征，原因不明。我的初步研究结果表明富含透明质酸的局部合成。细胞外基质（ECM）在鳞状细胞癌的发展中很重要，在拟议的研究中，我将确定它是如何发挥作用的。 ECM 有助于在斑马鱼的 SCC 形态发生中产生机械力（Aim1）。单细胞测序来识别参与活性 ECM 模式的基因 (Aim2) 并识别和描述 SCC 发展中涉及的其他 ECM 成分的作用 (Aim3)。这项研究揭示的力学原理将广泛适用于涉及其他形态发生事件此外，由于这些过程在许多情况下被破坏，因此活跃的 ECM 和拓扑上皮重塑。先天性疾病和耳部疾病，我的发现将提高我们对这些疾病的认识。拟议的研究借鉴了 1) 我在开发背景下进行的定量细胞生物学培训，2) 新的我在博士后研究中获得的技能，包括斑马鱼研究和发育器官成像单细胞分辨率，以及 3) 我的 K99 培训计划，用于实施自动图像分析、体内生物物理为了实现我的目标，我将接受指导。来自 Sean Megason、Tim Mitchison、L. Mahadevan 和 Allon Klein 博士，他们在斑马鱼方面拥有丰富的专业知识开发、基因编辑、生物物理学、定量成像、建模和单细胞 RNA 测序将提供我需要接受培训来开始我的独立研究计划，以促进我的职业发展。在指导的K99阶段，我将学习相关课程并保持基础知识的合作和转化研究，在会议上展示我的工作，发表我的研究，撰写补助金，并获得进一步的成果管理和指导学生的经验对于制定我自己的指导风格将受益匪浅。根据我的合作导师的经验、跟踪记录和定期反馈，我开始了自己的实验室的创建。我的长期职业目标是领导一个跨学科研究项目，研究机械和形态发生的分子基础，并利用我所获得的见解来推进我所做的转化研究。在研究经验、成功的合作和 K99 奖将使我能够获得成功所需的额外培训和专业知识。向独立过渡。