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迁移的主要细胞粘附分子。在粘附过程中，整联蛋白是 聚集以形成各种复合物，包括新生的粘附，小焦点络合物和成熟的局灶性粘连。 通过这些位点，细胞在底层上施加张力。我们对粘附组装的了解的大部分 复合物来自对细胞外基质（ECM）配体的细胞的研究。这个细胞扩散 测定对于理解粘附形成的机制非常有力，但并未完全概括关键特征 和细胞迁移过程中粘附形成的行为。特别是，它不考虑两极分化的形成 细胞前部附近的粘附，并且没有考虑到整联蛋白的内吞回收，这很关键 用于在2D和3D环境中的细胞迁移。我们已经开发了一种新的焦点粘附组件测定法 至关重要的取决于从粘连中整联蛋白的内吞作用得出的再生整合素。有趣的是，这个整合素 在非配合和主动构象中行驶，并导致高度极化的粘附形成。 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 回收整合素的假设是在其前缘附近的胞吞作用附近播种新的粘附形成。 我们的三个目的是：1）检查整联蛋白回收，激活状态，外旋转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附的极化形成 再生整联蛋白和3）确定细胞外基质拓扑和刚度如何影响形成的粘附形成 再生整联蛋白。在整个过程中，我们将研究回收整合素的粘附形成如何影响能力 细胞迁移。这项工作将促进细胞用来对其粘附的基本机制的理解 细胞迁移，这可能会导致细胞迁移不受监管的情况下的新策略，例如 癌症转移和持续性炎症。