Excitable Networks in Directed Cell Migration

定向细胞迁移中的兴奋网络

• 批准号: 10819960
• 负责人: Peter N Devreotes
• 金额: $ 5万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10819960
• 关键词: Acute; Address; Adult; Back; Biological; Biological Assay; Cell Death Induction; Cell Line; Cells; Charge; Chemotactic Factors; Coupled; Cytoskeleton; Dictyostelium; Disease; Embryo; Epithelial Cells; Face; Grant; Health; Image; Lipids; Location; Macrophage; Malignant Neoplasms; Mammalian Cell; Mediating; Membrane; Methods; Modeling; Molecular; Monitor; Normal Cell; Organoids; Pathology; Pattern; Phagosomes; Phosphotransferases; Physiology; Process; Property; Protein Geranylgeranylation; Proteins; Series; Signal Transduction; Starvation; Surface; System; Vesicle; cancer cell; cell behavior; cell motility; cell transformation; computer studies; design; genetically modified cells; inhibitor; migration; neutrophil; novel; novel therapeutic intervention; optogenetics; response; screening; spatiotemporal; tool; Acute; Address; Adult; Back; Biological; Biological Assay; Cell Death Induction; Cell Line; Cells; Charge; Chemotactic Factors; Coupled; Cytoskeleton; Dictyostelium; Disease; Embryo; Epithelial Cells; Face; Grant; Health; Image; Lipids; Location; Macrophage; Malignant Neoplasms; Mammalian Cell; Mediating; Membrane; Methods; Modeling; Molecular; Monitor; Normal Cell; Organoids; Pathology; Pattern; Phagosomes; Phosphotransferases; Physiology; Process; Property; Protein Geranylgeranylation; Proteins; Series; Signal Transduction; Starvation; Surface; System; Vesicle; cancer cell; cell behavior; cell motility; cell transformation; computer studies; design; genetically modified cells; inhibitor; migration; neutrophil; novel; novel therapeutic intervention; optogenetics; response; screening; spatiotemporal; tool

We are investigating molecular mechanisms of directed cell migration, a critical process in health and disease, using Dictyostelium as a discovery tool to inform our studies of neutrophils, macrophages, and epithelial cells. At the core of our working model are coupled Signal Transduction and Cytoskeletal Excitable Networks, referred to as STEN and CEN which drive motility. The STEN integrates inputs from directional sensing and polarity networks to bring about directed migration. In the last grant period, we found that protrusions are governed by waves of coupled STEN-CEN activities and that manipulation of negatively charged lipids on the inner face of the membrane can alter network excitability and control cell behavior. The STEN-CEN concept is conserved in mammalian cells and the networks are hyperactivated in transformed cells, augmenting motility and macropinocytosis. Since these processes require geranyl geranylation, statins cause starvation of cancer cells. Finally, we found that vesicles internalized from retracting protrusions carry “back” components to the rear of the cell contributing to polarity. How do diverse cellular protrusions depend on the setpoint/threshold of STEN-CEN? We are combining imaging, synthetic biological, and computational studies to prove that pseudopods, lamellipods, forming phagosomes, and so on are closely related on a spectrum and interconvertible. We will show that spatiotemporal patterns of activities and responses to acute molecular perturbations are consistent across these protrusions and parallel those established in propagating STEN-CEN waves? What explains the extraordinary coordination of activities in STEN and CEN? Surmising that charge on the inner leaflet of the membrane is an organizer, we are 1) designing methods to directly monitor charge in local regions; 2) determining how anionic lipids transiently decrease; 3) manipulating charge locally with optogenetic systems; 4) examining how the location of key proteins is regulated by surface charge. Is lowered STEN threshold a general property of transformed cells and can it be exploited? To address this question, we are 1) comparing threshold indicators, such as propagating waves, with macropinocytosis and statin sensitivity in a series of increasingly metastatic cell lines and organoids; 2) identifying the essential geranylgeranylated proteins in STEN; 3) genetically engineering cells to increase threshold to normalize cancer cells or further decrease threshold to induce cell death. How does control of STEN and CEN at the cell poles mediate directional sensing and polarity? First, using a novel suppression assay, we are screening kinase and substrate deficient cells to identify global inhibitors. Second, we are studying membrane flow in a variety conditions to pursue our “reverse fountain” model and reconcile with alternate models that argue membrane flows from front to back. 1

我们正在研究定向细胞迁移的分子机制，这是健康和 疾病，使用dictyostelium作为发现工具，以告知我们对中性粒细胞，巨噬细胞和 上皮细胞。我们工作模型的核心是耦合信号转导和细胞骨架 可兴奋的网络，称为sten和cen，可驱动运动。来自 定向感应和极性网络，以实现定向迁移。在最后一个赠款期，我们 发现突出受到耦合的sten-cen活性波的控制，并操纵 膜内面部负电荷的脂质会改变网络令人兴奋并控制细胞 行为。 Sten-Cen概念在哺乳动物细胞中保守，网络过度激活 在转化的细胞中，增强运动性和大型细胞增多症。由于这些过程需要香肠 天烷基化，他汀类药物会导致癌细胞的饥饿。最后，我们发现蔬菜从 缩回的突起载有“背部”成分，导致细胞的后部产生极性。 不同的细胞蛋白如何取决于Sten-Cen的设定点/阈值？我们是 结合成像，合成生物学和计算研究，以证明伪足 薄片，形成的吞噬体等在频谱上与互连密切相关。 将表明活动的时空模式和对急性分子扰动的响应是 在这些蛋白质中保持一致，并平行于传播sten-cen波的建立的蛋白质？ 是什么解释了Sten和CEN活动的非凡协调？推测这笔费用 膜的内部小叶是一个组织者，我们是1）设计直接监控电荷的方法 在地方地区； 2）确定阴离子脂质如何瞬时降低； 3）当地操纵电荷 使用光遗传系统； 4）检查如何通过表面电荷调节关键蛋白的位置。 是否降低了Sten阈值转化的细胞的一般特性，可以探索它吗？解决 这个问题，我们是1）将阈值指标（例如传播波）与 一系列越来越多的转移性细胞系和类器官中的大型细胞增多症和他汀类药物的敏感性； 2） 鉴定Sten中必需的黄烷基化蛋白； 3）基因工程细胞增加 阈值使癌细胞归一化或进一步降低阈值以诱导细胞死亡。 细胞杆处的Sten和CEN的控制如何介导定向感应和极性？第一的， 使用新型的抑制测定法，我们正在筛选激酶和底物特定细胞以识别全局 抑制剂。其次，我们正在研究各种条件下的膜流程，以追求“反向 喷泉”的模型，并与替代模型进行调和，这些模型反对膜从正面到背面流动。 1