Endogenous stem cells in jaw joint cartilage and ligament regeneration

项目摘要 常驻组织特异性干细胞用于修复和维持许多成人器官;然而，哺乳动物的关节 软骨和韧带具有非常差的内在修复能力。缺乏强大的内源性修复 可能是许多关节退行性疾病，特别是骨关节炎的根源。Smeeton医生 南加州大学盖奇·克博士实验室的博士后工作使用高度再生的斑马鱼来研究 干细胞在重建颌关节软骨和韧带中的作用。与哺乳动物一样，手术损伤 支撑斑马鱼颌关节的韧带导致关节软骨的损失。然而， 与哺乳动物不同的是，斑马鱼的韧带和关节软骨随后都再生。 本提案中概述的目标将建立在Smeeton博士广泛的初步数据基础上， 手术模型，并利用新的转基因和CRISPR/Cas9敲入系来研究时间轴 和颌骨关节再生的主要细胞事件（目的1），Sox 9+和Sox 10+茎的各自作用 细胞重建韧带和软骨（目标2），以及分子事件驱动再生（目标3）。目的 K99阶段将完成1个。目标2和3将在K99阶段的指导下启动 Crump博士和她的咨询委员会的报告，然后在R 00阶段完成。从长远来看， 从研究内源性Sox 9/10干细胞在斑马鱼颌关节中的作用中获得的信息可以被利用， 来唤醒哺乳动物关节中的类似细胞群以改善修复。 本建议书中概述的项目和指导计划旨在为博士奠定基础。 斯米顿的独立研究项目。在博士后任期结束后，她的目标是获得终身教职 顶级学术研究机构的助理教授职位。利用她新颖的手术模型， 最先进的遗传学和成像方法在斑马鱼，她的目标是使用斑马鱼颌骨关节，以了解如何 为了更好地从常驻干细胞重建软骨和韧带-一个与人类明显相关的主题 健康在K99阶段，她的科学和职业发展都将受益于以下方面的专业知识： 她的导师克博士研究斑马鱼骨骼发育和再生。此外，还定期与 她顾问委员会将使她获得关节生物学（丹尼斯·埃夫先科）、韧带 (Amy梅里尔）和RNAseq方法（Andrew McMahon）。南加州大学的其他职业发展活动， 如赠款写作，演示和指导讲习班将准备博士斯米顿她的过渡到一个 在R 00阶段的教师职位。麦克马洪博士将是一个特别重要的顾问过渡到 考虑到他在哈佛和南加州大学指导教师搜索的丰富经验，与其 广泛的颅面生物学家和软骨研究人员网络， 发展，和优秀的技术资源，南加州大学是一个充满活力和令人兴奋的地方建立博士。 斯米顿在关节和颅面再生方面的职业生涯。

Project Summary Resident tissue-specific stem cells serve to repair and maintain many adult organs; however mammalian joint cartilage and ligaments have a very poor capacity for intrinsic repair. This lack of robust endogenous repair may be at the root of many degenerative diseases of joints, in particular osteoarthritis. Dr. Smeeton's postdoctoral work in Dr. Gage Crump's lab at USC uses the highly regenerative zebrafish to investigate the role of stem cells in rebuilding the cartilage and ligaments of the jaw joint. As in mammals, surgical injury to the ligament supporting the zebrafish jaw joint results in loss of articular cartilage. However, a remarkable difference from mammals is that both the ligament and joint cartilage subsequently regenerate in zebrafish. The aims outlined in this proposal will build on Dr. Smeeton's extensive preliminary data establishing this new surgical model and make use of novel transgenic and CRISPR/Cas9 knock-in lines to investigate the timeline and major cellular events of jaw joint regeneration (Aim 1), the respective roles of Sox9+ and Sox10+ stem cells in rebuilding ligaments and cartilage (Aim 2), and the molecular events driving regeneration (Aim 3). Aim 1 will be completed in the K99 phase. Aims 2 and 3 will be initiated during the K99 phase under the mentorship of Dr. Crump and her advisory committee and then completed during the R00 phase. In the long term, insights gained from studying the role of endogenous Sox9/10 stem cells in the zebrafish jaw joint could be harnessed to awaken analogous cell populations in mammalian joints to improve repair. The project and mentorship plan outlined in this proposal were designed to lay the groundwork for Dr. Smeeton's independent research program. Following her postdoctoral term, her goal is to obtain a tenure-track Assistant Professor position at a top-tier academic research institute. Using her novel surgical models and state-of-the-art genetics and imaging methods in zebrafish, her goal is use the zebrafish jaw joint to learn how to better rebuild cartilage and ligaments from resident stem cells – a topic with a clear relevance for human health. During the K99 phase, both her scientific and career development will benefit from the expertise in zebrafish skeletal development and regeneration of her mentor, Dr. Crump. In addition, regular meetings with her advisory committee will allow her to acquire new expertise in joint biology (Denis Evseenko), ligaments (Amy Merrill), and RNAseq methodology (Andrew McMahon). Additional career development activities at USC, such as grant-writing, presentation and mentorship workshops will prepare Dr. Smeeton for her transition to a faculty position in the R00 phase. Dr. McMahon will be a particularly important advisor for the transition to independence given his extensive experience directing faculty searches at Harvard and USC. With its extensive network of craniofacial biologists and cartilage researchers, institutional support for career development, and outstanding technical resources, USC is a vibrant and exciting place to establish Dr. Smeeton's career in joint and craniofacial regeneration.