BEG4/MIM FUNCTION IN EPITHELIAL NEOPLASIA

描述(申请人提供)：尽管引导细胞迁移在正常皮肤发育和伤口愈合中起着关键作用，但引导伤口闭合或皮肤发育的细胞生物学机制仍然知之甚少。我们和其他人以前已经证明，转移中缺失(MIM)是一类新的膜相关BAR(I-BAR)结构域蛋白的定义成员，它调节肌动蛋白细胞骨架。我们已经利用果蝇边缘细胞迁移提供了第一个遗传学证据，证明I-bar蛋白通过一种新的依赖于表皮生长因子的导向机制来调控定向细胞迁移。我们的工作假设是亲内吞/CD2AP/Cortactin促进内吞作用和迁移，而抗内吞蛋白MIM在空间上抑制内吞作用，在皮肤发育和伤口愈合过程中提供定向信号。BIRT修订中提出的研究将父母资助的研究扩展到皮肤的迁移系统，并专注于识别与MIM一起工作的更多蛋白质来指导细胞迁移。我们将与斯坦福大学杰出的系统生物学家、高通量细胞成像分析专家托拜厄斯·迈耶教授合作。Oro-Meyer研究团队将独一无二地适合实施这一创新方法。这项合作建立在我们在发育遗传学方面的专业知识，以及迈耶实验室在活细胞成像和siRNA筛选方面的专业知识。我们将首先确定哪个特征良好的迁移系统使用依赖于MIM的转向。接下来，使用高通量迁移分析和细胞跟踪算法，我们将进行全基因组siRNA筛选，寻找像MIM这样补充皮质蛋白丢失的基因。我们的里程碑是识别使用MIM引导其迁移线索的脊椎动物细胞类型，并在siRNA筛选中识别10个基因，以便进一步用我们的体内分析进行验证。识别依赖MIM的导向机制的新组件将为理解细胞在发育过程中如何正常移动到其所需位置以及肿瘤细胞在癌症转移过程中如何使用类似机制提供关键见解。

DESCRIPTION (provided by applicant): While guided cell migration plays a critical role in normal skin development and wound healing, the cell biological mechanisms that guide wound closure or the developing skin remain poorly understood. We and others have previously shown that Missing in Metastasis (MIM) is a defining member of a novel class of membrane associated BAR (I-BAR) domain protein that regulates the actin cytoskeleton. We have used Drosophila border cell migration to provide the first genetic evidence that I-BAR proteins regulate directed cell migration through a novel epidermal growth factor-- dependent steering mechanism. Our working hypothesis is that pro-endocytic endophilin/CD2AP/cortactin enhance endocytosis and migration, while the anti-endocytic protein MIM spatially inhibits endocytosis to provide a directional signal during skin development and wound healing. The studies proposed in the BIRT revision extend the parent grant studies to migratory systems in skin and focuses on identifying additional proteins that work with MIM to direct cell migration. We will team with Professor Tobias Meyer, a prominent systems biologist at Stanford and an expert on high-throughput cell imaging analysis. The Oro-Meyer research team will be uniquely suited to carry out this innovative approach. The collaboration is built on our expertise in the genetics of development, and the Meyer lab's expertise in live cell imaging and siRNA screening. We will first identify which well characterized migratory system uses MIM-dependent steering. Next, using high throughput migration assays and cell tracking algorithms, we will perform a genome wide siRNA screen for genes like MIM that complement loss of cortactin. Our milestones are to identify vertebrate cell types that use MIM to steer toward their migratory cues and to identify 10 genes in the siRNA screen for further validation with our in vivo assays. Identification of novel components of the MIM-dependent steering mechanisms will provide key insights into understanding of how cells normally move to their desired locations during development and how tumor cells may use similar mechanisms during cancer metastasis.