An investigation of the roles of mechanical signaling in YAP-mediated tooth renew

机械信号在 YAP 介导的牙齿更新中作用的研究

• 批准号: 9904599
• 负责人: Jimmy Kuang-Hsien Hu
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9904599
• 关键词: 3-Dimensional; Ablation; Actins; Actomyosin; Address; Adult; Advisory Committees; Affect; Area; Binding; Biology; Biomechanics; CDC42 gene; Cell Compartmentation; Cell Culture Techniques; Cell Differentiation process; Cell Proliferation; Cell Shape; Cell physiology; Cells; Cellular biology; Cervical; Chemicals; Chronic; Clinical; Collaborations; Color; Communities; Community Medicine; Culture Techniques; Data; Dental; Developmental Biology; Educational workshop; Epithelial; Epithelium; Extracellular Matrix; Extracellular Matrix Proteins; Focal Adhesion Kinase 1; Fostering; Foundations; Future; Genetic; Homeostasis; Human; Image; In Vitro; Incisor; Integrin Signaling Pathway; Integrins; Investigation; Knowledge; Laboratories; Lasers; Lead; Learning; Life Style; Light; Limb structure; Measures; Mechanics; Mediating; Mentors; Morphogenesis; Mus; Muscle Development; Mutant Strains Mice; Myosin Type II; Natural regeneration; Nonmuscle Myosin Type IIA; Nuclear; Organ; Pathway interactions; Patients; Pattern; Phase; Phosphotransferases; Play; Postdoctoral Fellow; Process; Proliferating; Proteins; Publishing; Regenerative Medicine; Regulation; Research; Research Personnel; Resources; Rodent; Role; Science; Signal Transduction; Solid; Spatial Distribution; Surveys; System; Techniques; Testing; Time; Tissues; Tooth Loss; Tooth structure; Training; Transcription Coactivator; Universities; Work; Writing; adult stem cell; base; blebbistatin; cell behavior; craniofacial; craniofacial structure; epithelial stem cell; experience; experimental study; gain of function mutation; high throughput screening; in vivo; interest; mechanotransduction; meetings; mouse genetics; non-muscle myosin; novel; progenitor; public health relevance; rho GTP-Binding Proteins; screening; self-renewal; skills; small molecule; stem cell biology; stem cell niche; stem cell proliferation; stem cells; three dimensional cell culture; two photon microscopy

 DESCRIPTION (provided by applicant): Dr. Hu is determined to become an independent researcher to investigate the mechanisms that regulate the morphogenesis and homeostasis of craniofacial structures and to apply that knowledge for translational purposes. CANDIDATE BACKGROUND: Dr. Hu did his graduate research in Dr. Clifford Tabin's laboratory at Harvard University, where he studied limb patterning and muscle development, and gained experiences in developmental biology techniques, mouse genetics, live imaging, and cell culture. Dr. Hu then joined Dr. Ophir Klein's laboratory at UCSF as a postdoctoral fellow to study the mechanisms that regulate mouse incisor stem cells, a great system for understanding stem cell-based renewal. In particular, he found YAP/TAZ as important factors for controlling stem cell proliferation and differentiation and his preliminary data suggest that mechanical signaling plays a key role in regulating YAP activity and stem cell biology. During this time, Dr. Hu gained knowledge in craniofacial and stem cell biology, and has embarked on using the incisor as a system to study mechanotransduction, while developing techniques for culturing, imaging, and measuring force. RESEARCH PLAN: Dr. Hu hypothesizes that nuclear YAP localization and YAP-mediated cellular processes are initially inhibited by actomyosin tension in the incisor stem cells and later upregulatd by integrin/FAK signaling in the transit-amplifying cells. Aim 1 (K99 portion) studies the role of cellular tension by first characterizing cellular features affected by tension, such as cell shapes, levels of phospho-Myosin II, and actin distribution in the stem cell compartment. The effects of tissue tension on cell biology and YAP localization will be examined by laser ablation, and conditional deletion of Myosin IIA/B. Aim 2 (R00 portion) focuses on integrin/FAK signaling. First, the ECM compositions in the stem cell niche will be characterized. Functional studies will then be carried out by testing the ability of ECM proteins t regulate YAP activity and cell differentiation in a 3D culture system, as well as by examining two mouse mutants, one with FAK deletion and the other with a dominant active integrin β1. Finally, a chemical screen will be performed to identify novel factors and pathways that regulate YAP and stem cell biology. Together, these Aims will address key questions in both the Hippo and stem cell field. TRAINING: Dr. Hu will learn several new techniques during the mentored phase, including laser ablation, 3D culturing, mouse knock-ins, and high throughput small molecule screening. He will also attend courses relevant to the study. In addition, Dr. Hu has assembled an advisory committee to help complete and evaluate the project. Finally, he will continue to develop his writing, presentation, managing, and mentoring skills in order to become an independent researcher. ENVIRONMENT: There are many resources and researchers in different fields at UCSF and the greater Bay area science community that are available to Dr. Hu. There are also many regular seminars, workshops, and meetings at UCSF that foster scientific interactions, sharing of ideas, collaborations, and learning opportunities. HEALTH RELATEDNESS: As adult humans don't have dental epithelial stem cells, completion of this study will provide genetic, biomechanical, and chemical targets for developing strategies to derive and maintain clinically safe dental stem cells that can be used to make replacement teeth for treating patients with tooth loss.

 描述（由适用提供）：HU博士确定成为一名独立的研究人员，以研究调节颅面结构的形态发生和稳态的机制，并将这些知识应用于翻译目的。候选人背景：Hu博士在哈佛大学的Clifford Tabin博士的实验室中进行了研究生研究，在那里他研究了肢体图案和肌肉发育，并获得了发育生物学技术，小鼠遗传学，实时成像和细胞培养的经验。 Hu博士随后加入了UCSF的Ophir Klein博士的实验室，作为博士后研究员研究调节小鼠切牙干细胞的机制，这是理解基于干细胞的续订的好系统。特别是，他发现YAP/TAZ是控制干细胞增殖和分化的重要因素，他的初步数据表明，机械信号传导在调节YAP活性和干细胞生物学方面起关键作用。在此期间，Hu博士获得了有关颅面和干细胞生物学的知识，并开始使用切牙作为系统来研究机械转导的系统，同时开发用于聚类，成像和测量力的技术。研究计划：HU博士假设核YAP定位和YAP介导的细胞过程最初受到切牙干细胞中的肌球蛋白张力的抑制，然后在过失症中的综合素/FAK信号传导上进行了过失。 AIM 1（K99部分）研究细胞张力的作用，首先表征受细胞特征的影响 张力，例如细胞形状，磷酸肌球蛋白II的水平以及干细胞室中肌动蛋白分布。组织张力对细胞生物学和YAP定位的影响将通过激光消融以及肌球蛋白IIA/b的条件缺失进行检查。 AIM 2（R00部分）着重于整合素/FAK信号传导。首先，将表征干细胞生态位的ECM组成。然后，将通过测试ECM蛋白T的能力来进行功能研究，以调节3D培养系统中的YAP活性和细胞分化，以及检查两个小鼠突变体，一个具有FAK缺失，另一种具有主动蛋白β1。最后，将进行化学筛选以确定调节YAP和干细胞生物学的新因素和途径。这些目标共同解决了河马和干细胞场中的关键问题。培训：HU博士将在修补阶段学习几种新技术，包括激光消融，3D培养，小鼠敲击和高吞吐量小分子筛选。他还将参加与研究相关的课程。此外，HU博士还组建了一个咨询委员会，以帮助完成和评估该项目。最后，他将继续发展自己的写作，演讲，管理和心理技能，以成为一名独立的研究人员。环境：HU博士可以使用的UCSF和大湾地区科学界的不同领域的许多资源和研究人员。在UCSF上还有许多定期的半手，研讨会和会议，可以促进科学互动，共享思想，协作和学习机会。健康相关性：由于成年人没有牙齿上皮干细胞，这项研究的完成将提供遗传，生物力学和化学靶标，以制定策略来得出和维持临床上安全的牙科干细胞，可用于替代牙齿治疗牙齿脱落患者。