Mechanisms of Oral Epithelial Differentiation

摘要 此应用程序是指导临床科学家研究职业发展奖（K 08）博士凯文 伯德他目前正在进行研究有丝分裂纺锤体方向在口腔上皮细胞中的作用， 发展，并已观察到，在口腔粘膜上皮发育，垂直分裂， 变得越来越多，并且对分化至关重要。这款K 08将使伯德博士1）成为一名 通过有丝分裂纺锤体定位的分子调节口腔上皮分化的专家，2）进行 基础科学研究在体内的组织很少研究，3）获得技能，利用离体器官型 模型来描述人类口腔上皮分化中的这种现象，4）获得必要的技能， 在口腔上皮干细胞的鉴定中进行“组学”方法的合作，以及5）开发一种 独立的科学研究生涯，这将使他能够提出问题，与临床和基础 科学家一个多学科专家小组已同意在资助 本申请：1）Eric埃弗雷特博士（基础科学;牙科研究），Susan Henning博士（基础科学; 胃肠道研究），史蒂芬奥芬巴赫（转化和临床科学，牙科研究，和博士。 Ophir Klein（临床科学家：GI和牙科干细胞），2）上皮细胞和口腔生物学研究培训 专家：主要导师Scott威廉姆斯博士和共同导师Jennifer Webster-Cyriaque博士，以及3）RNA-seq 和生物信息学培训：（Piotr Mieczkowski，Scott Magness和Di Wu博士）。这些导师和 他们提供的培训将使他具备成为一名成功的独立调查员的技能。 口腔上皮小鼠模型的缺乏导致了口腔上皮细胞的特征化很少， 分化网络，以及对口腔上皮干细胞及其小生境的了解不足。基于他 快速生成（共8-12周）口腔上皮细胞小鼠模型，Byrd博士已经表明， 有丝分裂纺锤体定向基因LGN（Gpsm 2）在促进分化和组织形成中是必需的， 口腔。他可以产生新的口腔上皮小鼠模型的速度和特异性， 与传统方法相比，几周而不是几个月或几年的时间是前所未有的。伯德医生的 研究将集中在口腔上皮分化，这是理解口腔愈合的关键 以应对伤害。该项目将创建三种新型的口腔上皮小鼠模型和体外替代物 评估定向细胞分裂在口腔上皮分化中的作用的人类模型（目标1和2），以及 将利用无偏见的，遗传标记保留细胞模型，以确定和分析静止细胞从口腔基底 上皮细胞（Aim 3）。伯德博士的研究将为未来的储备干细胞功能项目提供依据 口腔上皮伤口愈合，将在资助期的最后一年提出（K99/R 00）。

ABSTRACT This application is for a Mentored Clinical Scientist Research Career Development Award (K08) for Dr. Kevin Byrd. He is currently conducting research into the role of mitotic spindle orientation in oral epithelial development and has observed that during oral mucosal epithelial development, perpendicular divisions become more numerous and are essential for differentiation. This K08 will allow Dr. Byrd to 1) become an expert in oral epithelial differentiation through molecular regulation of mitotic spindle positioning, 2) to conduct basic science research in vivo in a tissue rarely studied, 3) to gain the skills to utilize ex vivo organotypic models to describe this phenomenon in human oral epithelial differentiation, 4) to gain the skills necessary to collaborate and to perform “-omics” approaches in oral epithelial stem cell identification, and 5) to develop an independent science research career that will allow him to ask questions in collaboration with clinical and basic scientists. A multidisciplinary team of experts have agreed to provide invaluable support to him upon funding of this application: 1) Drs. Eric Everett (basic science; dental research), Susan Henning (basic science; gastrointestinal research), Steven Offenbacher (translational and clinical science, dental research, and Dr. Ophir Klein (clinician-scientist: GI and dental stem cells), 2) research training by epithelia and oral biology experts: primary mentor, Dr. Scott Williams and co-mentor, Dr. Jennifer Webster-Cyriaque, and 3) RNA-seq and bioinformatics training: (Drs. Piotr Mieczkowski, Scott Magness, and Di Wu). These mentors and the training they make available will provide him with the skills to become a successful, independent investigator. The paucity of oral epithelial mouse models has resulted in minimal characterization of oral epithelial differentiation networks, and a poor understanding of oral epithelial stem cells and their niches. Based on his rapid generation (8-12 weeks total) of an oral epithelia mouse model, Dr. Byrd has already shown that the mitotic spindle orientation gene LGN (Gpsm2) is essential in promoting differentiation and tissue patterning in the oral cavity. The pace and specificity in which he can generate new oral epithelial mouse models where few exist—weeks as opposed to months or years—is unprecedented compared to traditional methods. Dr. Byrd's research will focus on oral epithelial differentiation, which is critical in understanding how the oral cavity heals in response to injury. This project will create three novel, oral epithelial mouse models and surrogate ex vivo human models to assess the role of oriented cell divisions in oral epithelial differentiation (Aim 1 and 2), and will utilize a unbiased, genetic label retaining cell model to identify and profile quiescent cells from oral basal epithelial cells (Aim 3). Dr. Byrd's research will inform the basis of a future project on reserve stem cell function in oral epithelial wound healing, to be proposed in the last year of the funding period (K99/R00).