Endogenous stem cells in jaw joint cartilage and ligament regeneration

项目摘要 驻留的组织特异性干细胞用于修复和维持许多成年器官；然而哺乳动物关节 软骨和韧带的内在修复能力很差。缺乏强大的内源性修复 可能是许多退行性关节疾病的根源，特别是骨关节炎。斯米顿医生的 南加州大学盖奇·克伦普博士实验室的博士后研究人员使用高度再生的斑马鱼来研究 干细胞在重建颌骨关节软骨和韧带中的作用。就像在哺乳动物中一样，外科手术对 支持斑马鱼颌骨关节的韧带会导致关节软骨的丧失。然而，值得注意的是， 与哺乳动物的不同之处在于，斑马鱼的韧带和关节软骨随后都会再生。 这项提案中概述的目标将建立在斯米顿博士广泛的初步数据基础上，建立这一新的 手术模型和利用新的转基因和CRISPR/Cas9敲入系来研究时间线 和颌骨关节再生的主要细胞事件(目标1)，Sox9和Sox10茎各自的作用 重建韧带和软骨中的细胞(目标2)，以及驱动再生的分子事件(目标3)。目标 1个将在K99阶段完成。目标2和目标3将在K99阶段的指导下启动 克伦普博士和她的顾问委员会，然后在R00阶段完成。从长远来看，洞察力 通过研究内源性Sox9/10干细胞在斑马鱼颌骨关节中的作用获得的信息可以被利用 唤醒哺乳动物关节中类似的细胞群，以改善修复。 该提案中概述的项目和指导计划旨在为Dr。 斯米顿的独立研究计划。在博士后任期结束后，她的目标是获得终身教职 一家顶级学术研究机构的助理教授职位。使用她的新手术模型和 最先进的斑马鱼遗传学和成像方法，她的目标是利用斑马鱼的下颌关节来学习如何 更好地利用常驻干细胞重建软骨和韧带--这是一个与人类明显相关的话题 健康。在K99阶段，她的科学和职业发展都将受益于 斑马鱼的骨骼发育和再生，她的导师克伦普博士。此外，定期与 她的顾问委员会将允许她在关节生物学(Denis Evseenko)、韧带方面获得新的专业知识 (艾米·梅里尔)和RNAseq方法(安德鲁·麦克马洪)。南加州大学的其他职业发展活动， 例如，赠款撰写、演示和指导研讨会将为斯米顿博士过渡到 R00阶段的教师职位。麦克马洪博士将是过渡到 考虑到他在哈佛和南加州大学指导教员搜索的丰富经验，他是独立的。使用它的 广泛的颅面生物学家和软骨研究人员网络，为职业生涯提供机构支持 南加州大学是一个充满活力和令人兴奋的地方，是建立Dr。 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.