Excitable Networks in Directed Cell Migration

定向细胞迁移中的兴奋网络

• 批准号: 10187811
• 负责人: Peter N Devreotes
• 金额: $ 108.08万
• 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/10187811
• 关键词: Acute; Address; Adult; Back; Biological; Biological Assay; Cell Death; Cell Line; Cells; Charge; Chemotactic Factors; Coupled; Dictyostelium; Disease; Embryo; Epithelial Cells; Face; Grant; Health; Image; Lead; Lipids; Location; Malignant Neoplasms; Mammalian Cell; Mediating; Membrane; Methods; Modeling; Molecular; Monitor; Normal Cell; Organoids; Pathology; Pattern; Phagosomes; Phosphotransferases; Physiology; Process; Property; Proteins; Series; Signal Transduction; Starvation; Surface; System; Vesicle; cancer cell; cell behavior; cell motility; cell transformation; computer studies; design; genetically modified cells; inhibitor/antagonist; macrophage; 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 it 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，这是健康和健康的一个关键过程。 疾病，使用盘基网柄菌作为发现工具，为我们对中性粒细胞、巨噬细胞和 上皮细胞。我们工作模型的核心是信号转导和细胞骨架耦合 兴奋网络，称为 STEN 和 CEN，可驱动动力。 STEN 集成了来自 定向传感和极性网络带来定向迁移。在上一个资助期内，我们 发现突起是由耦合的 STEN-CEN 活动波控制的，并且操纵 膜内表面带负电荷的脂质可以改变网络兴奋性并控制细胞 行为。 STEN-CEN 概念在哺乳动物细胞中是保守的，并且网络是高度活跃的 在转化细胞中，增强运动性和巨胞饮作用。由于这些过程需要香叶基 香叶基化，他汀类药物会导致癌细胞饥饿。最后，我们发现囊泡内化自 缩回的突起将“返回”组件带到电池的后部，从而产生极性。 不同的细胞突起如何取决于 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来证明伪足， 片足类、形成吞噬体等在谱系上密切相关并且可以相互转换。我们 将表明活动的时空模式和对急性分子扰动的反应是 这些突起之间的一致性与传播 STEN-CEN 波中建立的平行突起一致。 如何解释 STEN 和 CEN 活动的非凡协调？推测充电 膜的内部小叶是一个组织者，我们正在1）设计直接监测电荷的方法 在当地地区； 2)确定阴离子脂质如何瞬时减少； 3）本地操纵电荷 与光遗传学系统； 4) 研究表面电荷如何调节关键蛋白质的位置。 降低的 STEN 阈值是转化细胞的一般特性吗？可以利用吗？致地址 这个问题，我们 1) 比较阈值指标，例如传播波，与 一系列日益转移的细胞系和类器官中的巨胞饮作用和他汀类药物敏感性； 2） 鉴定 STEN 中必需的香叶基香叶基化蛋白； 3）基因工程细胞增加 使癌细胞正常化的阈值或进一步降低诱导细胞死亡的阈值。 细胞两极的 STEN 和 CEN 控制如何介导方向传感和极性？第一的， 使用新型抑制测定，我们正在筛选激酶和底物缺陷细胞，以识别全局 抑制剂。其次，我们正在研究各种条件下的膜流，以追求我们的“反向流动” 喷泉”模型，它与认为膜从前向后流动的替代模型相一致。 1