Incisor stem cell dynamics in homeostasis and repair

门牙干细胞动态平衡和修复

• 批准号: 9163481
• 负责人: Amnon Sharir
• 金额: $ 18.21万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9163481
• 关键词: Ablation; Address; Adult; Affect; Ameloblasts; Animal Hospitals; Applications Grants; Area; Award; Biological Assay; Biological Models; Biology; Biomechanics; Biomedical Engineering; Candidate Disease Gene; Cell Cycle; Cell Proliferation; Cell Therapy; Cell physiology; Cells; Cervical; Clinical; Collaborations; Communication; Data; Dental; Dental caries; Development; Developmental Biology; Devices; Distal; Engineering; Epithelial; Epithelium; Equilibrium; Event; Fluorouracil; Foundations; Frequencies; Future; Genes; Genetic; Genetic screening method; Goals; High Prevalence; Homeostasis; Human; Human Engineering; Incisor; Individual; Institutes; Israel; Knowledge; Label; Laboratories; Learning; Length; Life; Location; Maintenance; Mechanics; Mentors; Modeling; Molecular; Monoclonal Antibodies; Mus; Muscle; Natural regeneration; Organ; Orthodontic; Pathway interactions; Patients; Pattern; Play; Process; Production; Proliferating; Recovery; Reporter; Research; Research Personnel; Resources; Role; Schools; Science; Signal Pathway; Signal Transduction; Stem cells; System; Therapeutic; Tissues; Tooth Loss; Tooth Movement; Tooth regeneration; Tooth structure; Training; Translating; Universities; Veterinary Medicine; Work; Writing; adult stem cell; base; bone; career; cell behavior; cell type; craniofacial; cytotoxic; experience; in vivo; interest; killings; meetings; notch protein; orofacial; programs; reconstitution; repaired; research study; self-renewal; skeletogenesis; skills; stem cell biology; tissue repair; tool; transcriptome sequencing

ABSTRACT CAREER GOALS: Dr. Sharir is determined to become an independent researcher, to investigate the molecular and biomechanical bases of mouse tooth regeneration and to apply this knowledge for translational purposes. CANDIDATE BACKGROUND: Dr. Sharir graduated from the Koret School of Veterinary Medicine in Israel and worked in a private animal hospital, where he gained clinical experience. He did his graduate research in the laboratories of Dr. Ron Shahar at The Hebrew University of Jerusalem and Dr. Elazar Zelzer at The Weizmann Institute of Science. He studied the role of muscle force in skeletogenesis and gained experience in developmental biology, genetics and bone mechanobiology. Dr. Sharir then joined Dr. Ophir Klein's laboratory at UCSF to study the mechanisms underlying the ability of stem cells (SC) to regenerate dental tissues, which is an excellent model system for in vivo exploration of SC behavior. In particular, Dr. Sharir studies the mechanisms that control the balance between self-renewal and differentiation of dental epithelium stem cells (DESCs). His preliminary findings question the common view of DESCs as slow-cycling, asymmetrically dividing cells, inspiring new directions to be explored in this proposal. RESEARCH PLAN: Dr. Sharir hypothesizes that the mouse incisor epithelium is maintained by a pool of actively cycling, equipotent SCs that are regulated by external mechanical forces. Aim 1 addresses the dynamics of clones derived from genetically labeled DESCs, and how these dynamics change following: 1) cytotoxic ablation of proliferating cells, or 2) pharmacologic disruption of Notch signaling. These experiments DESCs and will determine whether they are slow cycling and asymmetrically dividing, or actively-cycling and equipotent. In Aim 2, the role of mechanical forces in regulating DESC dynamics will be studied. First, the experienced by the mouse incisor will be modified by a device previously used in orthodontic tooth movement and the relationship between the applied load and DESC cycling dynamics will be examined. An RNA-seq analysis will be then performed to identify differentially regulated genes in the incisor epithelium under these load conditions. Finally, the functional role of these genes in incisor regeneration will be tested by genetic or pharmacologic approaches. This research will form the basis for further investigating the molecular and biomechanical bases of tooth regeneration, to be proposed in an R01 grant application during the fourth year of this award. will expose the location of mechanical loads TRAINING: Dr. Sharir is well-prepared to undertake this research. Nevertheless, to complete the project and achieve his long-term career goals, he will require further training in the application of quantitative tools for analyzing adult SC behavior and dental mechanobiology. To this end, Dr. Sharir has assembled a team of mentors and collaborators comprising of experts in these areas with the ability to help him obtain these critical skills, complete the project and evaluate progress. He will also attend courses relevant to these topics. In addition, through courses at UCSF and further hands-on experience, this award will help Dr. Sharir to develop his communication, writing and mentoring skills to eventually launch a fruitful independent research career. ENVIROMENT: Dr. Klein's lab and UCSF provide an intellectually stimulating, collaborative and resource-rich enviroment that is ideal for executing the proposed research and for promoting Dr. Sharir's career. The host lab is a world leader in the study of tooth SCs and is well-equipped for the proposed study. The Department of Orofacial Sciences and Program in Craniofacial Biology will provide additional resources for collaborations and learning opportunities.

摘要 职业目标：Sharir博士决心成为一名独立的研究人员， 和小鼠牙齿再生的生物力学基础，并将这些知识用于翻译目的。 候选人背景：Sharir博士毕业于以色列Koret兽医学院， 他在一家私人动物医院工作，在那里他获得了临床经验。他的研究生研究是在 耶路撒冷希伯来大学的罗恩·沙哈尔博士和魏茨曼大学的埃拉扎尔·泽尔泽博士的实验室 科学研究所。他研究了肌肉力量在骨骼形成中的作用， 发育生物学、遗传学和骨机械生物学。Sharir博士随后加入了Ophir Klein博士的实验室 研究干细胞（SC）再生牙齿组织能力的机制， 是一个很好的模型系统，在体内探索SC的行为。特别是，Sharir博士研究了 牙上皮干细胞自我更新与分化平衡的调控机制 （DESC）。他的初步研究结果质疑了DESC是缓慢循环、不对称的普遍观点， 分裂的细胞，启发了这个提议中要探索的新方向。 研究：Sharir博士假设，小鼠切牙上皮细胞是由一个池， 主动循环，等效SC由外部机械力调节。 目标1针对 从遗传标记的DESC衍生的克隆的动态，以及这些动态如何改变如下：1） 增殖细胞的细胞毒性消融，或2）Notch信号传导的药理学破坏。这些实验 DESC，并将确定它们是否是缓慢循环和不对称的 分裂，或主动循环和等势。在目标2中，机械力在调节DESC中的作用 将研究动态。首先，将鼠标所经历的门齿进行修改， 以前用于正畸牙齿移动的装置以及施加的载荷与 DESC循环动力学将被检查。然后将进行RNA-seq分析以鉴定差异 在这些负荷条件下切牙上皮中的调节基因。最后，这些功能的作用 将通过遗传学或药理学方法测试切牙再生中的基因。这项研究将形成 提出了进一步研究牙齿再生的分子和生物力学基础的基础 在R 01补助金申请在这个奖项的第四年。 会暴露 机械载荷 培训：Sharir博士为开展这项研究做好了充分的准备。然而，为了完成该项目， 为了实现他的长期职业目标，他将需要进一步的培训，在应用定量工具， 分析成年SC行为和牙齿机械生物学。为此，Sharir博士组建了一个团队， 导师和合作者，由这些领域的专家组成，有能力帮助他获得这些关键的 技能，完成项目并评估进度。他还将参加与这些主题有关的课程。在 此外，通过在UCSF的课程和进一步的实践经验，这个奖项将有助于博士。 他的沟通，写作和指导技能，最终推出了一个富有成效的独立研究生涯。 克莱因的实验室和加州大学旧金山分校提供了一个智力刺激，协作和资源丰富的 这是执行拟议的研究和促进Sharir博士的职业生涯的理想环境。主机 实验室是牙齿干细胞研究的世界领导者，并为拟议的研究配备了良好的设备。部 颅面科学和颅面生物学计划将为合作提供额外的资源， 学习机会。