Incisor stem cell dynamics in homeostasis and repair

摘要 职业目标：沙里尔博士决心成为一名独立的研究员，研究分子 以及小鼠牙齿再生的生物力学基础，并将这些知识应用于翻译目的。 候选人背景：沙里尔博士毕业于以色列科尔特兽医学院， 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Ron Shahar博士和魏茨曼的Elazar Zelzer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，并在 发育生物学、遗传学和骨骼机械生物学。Sharir博士随后加入了Ophir Klein博士的实验室 在加州大学旧金山分校研究干细胞(SC)再生牙齿组织能力的潜在机制 是在活体内探索SC行为的优秀模型系统。特别是，沙里尔博士研究了 牙上皮干细胞自我更新与分化平衡的调控机制 (DESCS)。他的初步发现对DESCs的普遍观点提出了质疑，认为DESCs是不对称的慢循环 分裂细胞，激发了在这项提案中探索的新方向。 研究计划：沙里尔博士假设小鼠的门牙上皮由一池 积极循环，等量的SC由外部机械力调节。 目标1针对的是 来自基因标记的DESCs的克隆的动力学，以及这些动力学如何变化如下：1) 细胞毒性消融增殖细胞，或2)药物干扰Notch信号。这些实验 DESCs，并将确定他们是否骑得很慢和不对称 除法，或积极循环和等价性。在目标2中，机械力在调节DESC中的作用 将对动力学进行研究。首先，由鼠标所经历的切牙将被修改为 用于正畸牙齿移动的装置及其与外加载荷的关系 将考察DESC自行车的动力学。然后进行rna-seq分析，以区分 在这些负荷条件下，切牙上皮中的调控基因。最后，这些功能的作用 门牙再生的基因将通过遗传学或药理学方法进行测试。这项研究将形成 为进一步研究牙齿再生的分子和生物力学基础提供依据 在本奖项第四年的R01拨款申请中。 将会暴露出 机械载荷 培训：沙里尔博士已经为开展这项研究做好了充分准备。尽管如此，为了完成该项目并 要实现他的长期职业目标，他将需要进一步培训量化工具的应用 成人SC行为和牙齿机械生物学分析。为此，沙里尔博士组建了一个团队， 由这些领域的专家组成的导师和合作者，能够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相关的课程。在……里面 此外，通过加州大学旧金山分校的课程和进一步的实践经验，该奖项将帮助Sharir博士发展 他的沟通、写作和指导能力最终开启了卓有成效的独立研究生涯。 环境：克莱恩博士的实验室和加州大学旧金山分校提供了一个智力激发、协作和资源丰富的环境 环境，这是执行拟议的研究和促进沙里尔博士的事业的理想环境。东道主 LAB是牙齿干细胞研究的世界领先者，并为拟议的研究提供了良好的设备。美国商务部 口腔面部科学和颅面生物学计划将提供更多的合作资源和 学习机会。

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.