Integrin Recycling and Adhesion Formation in Cell Migration

细胞迁移中整合素的回收和粘附形成

• 批准号: 9765849
• 负责人: Gregg G Gundersen
• 金额: $ 32.4万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2020-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9765849
• 关键词: 3-Dimensional; Accounting; Address; Adhesions; Affect; Behavior; Biochemical; Biological; Biological Assay; Cell Adhesion; Cell Adhesion Molecules; Cell membrane; Cells; Centrosome; Complex; Cytoskeleton; Development; Elements; Endocytosis; Environment; Exocytosis; Extracellular Matrix; Fluorescent Probes; Focal Adhesions; Hydrogels; Immune response; Inflammation; Integrins; Intervention; Kinesin; Lead; Ligands; Microtubules; Modeling; Molecular Conformation; Motor; Neoplasm Metastasis; Pattern; Post-Translational Protein Processing; Process; Property; Recycling; Role; Seeds; Site; Testing; Traction; Travel; Work; Wound Healing; biophysical properties; cell motility; design; high resolution imaging; imaging approach; novel; receptor; receptor binding; trafficking; virtual

Summary Integrins are the major cell adhesion molecules used by virtually all cells to migrate. During adhesion, integrins are clustered to form a variety of complexes including nascent adhesions, small focal complexes and mature focal adhesions. Through these sites, the cell exerts tension on the substratum. Much of what we know about the assembly of adhesion complexes comes from studies on cells detached and replated on extracellular matrix (ECM) ligands. This cell spreading assay has been powerful for understand mechanisms of adhesion formation, yet it does not fully recapitulate key features and behaviors of adhesion formation during cell migration. In particular, it does not account for the polarized formation of adhesions near the front of the cell and does not take into account the endocytic recycling of integrins, which is critical for cell migration in both 2D and 3D environments. We have developed a new focal adhesion assembly assay that is critically dependent on recycled integrin derived from the endocytosis of integrins in adhesions. Interestingly, this integrin travels in an unliganded and active conformation and leads to the highly polarized formation of adhesions at the leading edge of migrating cells. We will use cell biological, biochemical and high resolution imaging approaches to test the hypothesis that recycled integrin acts to seed new adhesion formation near sites of its exocytosis near the leading edge. Our three aims are: 1) to examine the relationship between integrin recycling, activation state, exocytic sites and adhesion formation, 2) to determine how the microtubule cytoskeleton contributes to the polarized formation of adhesions from recycled integrins and, 3) to determine how extracellular matrix topology and stiffness affect adhesion formation from recycled integrins. Throughout, we will examine how the formation of adhesions from recycled integrins affect the ability of cells to migrate. This work will advance understanding of the basic mechanisms cells use to polarize their adhesions for cell migration, which may lead to new strategies for intervention in cases where cell migration is unregulated, such as cancer metastasis and persistent inflammation.

概括 整合素是几乎所有细胞用于迁移的主要细胞粘附分子。在粘附过程中，整合素 聚集形成各种复合体，包括新生粘连、小斑复合体和成熟粘连斑。 通过这些位点，细胞对基质施加张力。我们所知道的关于粘合组装的大部分知识 复合物来自对细胞分离并重新铺在细胞外基质 (ECM) 配体上的研究。这种细胞扩散 测定对于理解粘附形成机制非常有效，但它并没有完全概括关键特征 以及细胞迁移过程中粘附形成的行为。特别是，它没有考虑极化形成 细胞前部附近的粘附，并且没有考虑整合素的内吞再循环，这是至关重要的 用于 2D 和 3D 环境中的细胞迁移。我们开发了一种新的粘着斑组装测定法 严重依赖于粘连中整合素内吞作用所产生的再循环整合素。有趣的是，这个整合素 以未配体且活跃的构象行进，并导致在前导处形成高度极化的粘附 迁移细胞的边缘。我们将使用细胞生物学、生化和高分辨率成像方法来测试 假设回收的整合素在其前缘附近的胞吐作用位点附近播种新的粘附形成。 我们的三个目标是：1）检查整合素回收、激活状态、胞吐位点和粘附之间的关系 形成，2) 确定微管细胞骨架如何促进粘连的极化形成 回收的整合素，3) 确定细胞外基质拓扑结构和硬度如何影响粘附形成 回收的整合素。在整个过程中，我们将研究回收的整合素形成的粘附如何影响能力 细胞迁移。这项工作将加深对细胞用于极化其粘附的基本机制的理解 细胞迁移，这可能会导致在细胞迁移不受调控的情况下进行新的干预策略，例如 癌症转移和持续炎症。