Investigating mechanisms of tissue polarity during development and disease

项目摘要 人类发育过程中的器官形成和组织模式依赖于集体和定向的细胞行为。 平面细胞极性（PCP），从苍蝇到人类保守的信号通路，管理这种协调 相邻细胞之间。PCP很好地代表了身体毛发的有序排列， 沿着前后体轴的哺乳动物皮肤。PCP蛋白功能缺陷导致严重的 发育缺陷，包括心肌病、纤毛病和神经管缺陷，如脊柱裂。 五氯苯酚通路中断如何导致发育障碍仍然知之甚少。重要的是，PCP 导致小鼠发育缺陷和胚胎死亡的破坏也导致不能正确地 因此，小鼠皮肤是一个合适的模型系统与毛囊极性 是PCP函数的易处理读出。PCP的一个定义特征是核心PCP的不对称定位 蛋白卷曲蛋白6（Fz 6）和货车高样蛋白2（Vangl 2）。细胞间相互作用 非典型钙粘蛋白Celsr 1是Fz 6和Vang 12在细胞边界定位所必需的。然而，我们不明白 Celsr 1粘附如何组织不对称PCP连接或在发育过程中如何调节。的 需要了解Celsr 1如何在连接处作为Fz 6-Vangl 2不对称性的驱动器， 最近在神经管患者中发现了新的、预测的致病性Celsr 1突变， 先天性心脏病这项提案的总体目标是更好地了解这些疾病相关的 Celsr 1突变，特别是那些映射到负责细胞粘附的Celsr 1结构域的突变，有助于 人类发育障碍以小鼠表皮为PCP功能的模型系统，沿着细胞 和分子生物学，生物化学，先进的光学成像和体内遗传方法，我将测试 假设疾病相关Celsr 1突变扰乱Celsr 1细胞粘附并改变 发育过程中PCP不对称。这一假设将在以下具体目标中进行探讨：1） 确定疾病相关的Celsr 1突变如何影响Celsr 1粘附和信号传导，以及2）确定 疾病相关的Celsr 1突变如何影响PCP的建立。拟议的目标将揭示 针对Celsr 1功能的发育障碍的病理机制，同时也提供了关键的见解， Celsr 1如何调节PCP的形成。奖学金支持，以完成拟议的研究和制作 培训计划，沿着我的指导团队的指导和宾夕法尼亚州立大学的环境， 医学，将有助于我的职业发展，朝着我领导一个独立和多样化的目标。 研究团队。

PROJECT SUMMARY Organ formation and tissue patterning during human development rely on collective and oriented cell behaviors. Planar cell polarity (PCP), a signaling pathway conserved from flies to humans, governs this coordination between neighboring cells. PCP is well represented by the ordered alignment of body hairs across the mammalian skin along the anterior-posterior body axis. Deficiencies in PCP protein function result in severe developmental defects including cardiomyopathies, ciliopathies and neural tube defects such as spina bifida. How PCP pathway disruption results in developmental disorders remains poorly understood. Importantly, PCP disruption resulting in developmental defects and embryonic lethality in mice also results in a failure to properly pattern the embryonic epidermis, thus the mouse skin is a suitable model system with and hair follicle polarity being a tractable read-out of PCP function. A defining feature of PCP is the asymmetric localization of core PCP proteins Frizzled 6 (Fz6) and Van Gogh like protein 2 (Vangl2) at cell-cell borders. Intercellular interactions of atypical cadherin Celsr1 is required for the Fz6 and Vangl2 localization at cell borders. Yet, we do not understand how Celsr1 adhesion organizes asymmetric PCP junctions or how this is regulated during development. The need to understand how Celsr1 functions as a driver of Fz6-Vangl2 asymmetry at junctions is underscored by the recent identification of novel, predicted pathogenic, Celsr1 mutations in patients with neural tube and congenital heart defects. The overall goal of this proposal is to better understand how these disease-associated Celsr1 mutations, particularly those that map to the Celsr1 domains responsible for cell adhesion, contribute to human developmental disorders. Using the murine epidermis as model system for PCP function, along with cell and molecular biology, biochemical, advanced optical imaging and in vivo genetic approaches, I will test the hypothesis that disease-associated Celsr1 mutations perturb Celsr1 cell adhesion and alter the establishment of PCP asymmetry during development. This hypothesis will be interrogated in the following specific aims: 1) Determine how disease-associated Celsr1 mutations impact Celsr1 adhesion and signaling, and 2) Determine how disease-associated Celsr1 mutations impact PCP establishment. The proposed aims will reveal pathomechanisms of developmental disorders that target Celsr1 function while also providing key insight into how Celsr1 regulates PCP establishment. Fellowship support to complete the proposed research and crafted training plan, along with guidance from my mentoring team and the environment at the Penn State College of Medicine, will be instrumental in my career development toward my goal of leading an independent and diverse research team.