Role of Wnt-responsive cells in oral mucosa homeostasis, injury, and malignancy

项目摘要/摘要 口腔粘膜容易出现创伤、先天缺陷和后天疾病(如癌症)等疾病。 同时，口腔粘膜是保护底层组织的屏障，这是通过 在基底层不断地替换来自假定的干细胞储存库的细胞。目前， 口腔粘膜中干细胞的特性在很大程度上是未知的，这阻碍了进一步的机制研究。 了解它们的自我更新、分化和再生能力。我开发了一套转基因技术的曲目 血统追踪小鼠，并确定了口腔中具有干细胞库功能的Wnt反应群体 上皮细胞。在这个拟议的项目中，我将彻底描述这些WNT响应茎的贡献(S) 口腔粘膜细胞在动态期(Aim1)、损伤修复期(Aim2)和恶变期(Aim3)。我的中心假设是- Esis是指在口腔粘膜中，Wnt反应细胞维持组织的完整性并实现再生 在受伤后，这一群体的突变会引发口腔癌。这些目标将利用血统追踪， 谱系消融，标记保留，损伤和口腔癌模型，以询问干细胞的维护，再生- 发情动力学以及癌症起源的细胞。这些研究旨在丰富我们的知识 关于干细胞在口腔粘膜中的分布以及这些细胞如何对口腔粘膜的完整性和愈合做出贡献 口腔粘膜的能力。这一未被研究的领域具有巨大的潜力，可以彻底改变治疗方法 颅面复合体组织的再生，包括皮肤和粘膜。成功完成这项工作 该项目还将促进我们对癌症干细胞的了解，为口腔癌的治疗提供新的靶点。 目标2和目标3将在吉尔·赫尔姆斯博士(斯坦福大学)丹尼尔·拉莫斯博士的指导下启动 (加州大学旧金山分校)和Sinha Satrajit博士(布法罗大学)，并在R00期间完成，而Aim 1将 在K99阶段结束前完成。这个项目旨在促进我的职业目标 在一家顶级学术研究机构获得终身教职助理教授的职位。我计划这样做 开发一个独立的研究计划，依靠最先进的遗传小鼠模型来解决未解决的问题 口腔粘膜中与人类健康明显相关的问题。除了专心致志的研究时间，我将利用 完成课程并参加研讨会，以更新我在快速发展的干细胞生物学和 斯坦福大学的癌症生物学，它拥有最有经验和最繁荣的社区之一 世界上癌症和干细胞生物学家。我还将学习高级生物统计学，以改进实验设计 和数据解释。为过渡到寻求R01级资金的独立调查员做准备， 我将从我的导师那里积极参与学习助学金的撰写、教学、指导和沟通技巧 和顾问，以及斯坦福大学提供的互动讲座和辅导课程。

Project Summary/Abstract Oral mucosa is prone to disorders such as trauma, congenital defects, and acquired diseases (e.g., cancer). Meanwhile, oral mucosa serves as a barrier to protect the underlying tissues, which is achieved through the continual replacement of cells originating from a presumable reservoir of stem cells in the basal layer. Currently, the identity of stem cells in oral mucosa is largely unknown, preventing further mechanistic studies to fully un- derstand their self-renewal, differentiation and regeneration capacity. I have developed a repertoire of transgenic lineage tracing mice and identified a Wnt-responsive population that functions as a stem cell pool in the oral epithelia. In this proposed project, I will thoroughly characterize the contribution(s) of these Wnt-responsive stem cells in oral mucosa during homeostasis (Aim1), injury repair (Aim2), and malignancy (Aim3). My central hypoth- esis is that in oral mucosa, Wnt-responsive cells maintain the tissue integrity and effectuate the regeneration following injury, and mutation of this same population initiates oral cancer. These aims will utilize lineage tracing, lineage ablation, label retention, and injury and oral cancer models to interrogate stem cell maintenance, regen- erative dynamics as well as the cells of origin in cancer. These studies are designed to enrich our knowledge about the distribution of stem cells in oral mucosa and how those cells contribute to the integrity and healing ability of oral mucosa. This understudied area has great potential to revolutionize the therapeutic approaches for regeneration of tissues in the craniofacial complex, including skin and mucosa. Successful completion of this project will also advance our knowledge of cancer stem cells, providing novel targets for oral cancer treatment. Aim 2 and 3 will be initiated under the mentorship of Dr. Jill Helms (Stanford University), Dr. Daniel Ramos (UCSF) and Dr. Sinha Satrajit (University at Buffalo) and completed during the R00 period, whereas Aim 1 will be completed before the end of the K99 phase. This project has been designed to facilitate my career goal of obtaining a position as a tenure-track Assistant Professor at a top-tier academic research institution. I plan to develop an independent research program relying on state-of-the-art genetic mouse models to address unsolved questions in oral mucosa with clear relevance to human health. Besides the dedicated research time, I will take coursework and attend seminars to update my knowledge in the rapidly evolving fields of stem cell biology and cancer biology at Stanford University which hosts one of the most experienced and thriving communities of cancer and stem cell biologists in the world. I will also learn advanced biostatistics to improve experiment design and data interpretation. To prepare for the transition into an independent investigator pursuing R01-level funding, I will be actively engaged in learning grant writing, teaching, mentoring and communication skills from my mentor and advisors, and also from the interactive lectures and coaching sessions offered from Stanford University.