Mechanisms for regulation of cell adhesion and migration

细胞粘附和迁移的调节机制

• 批准号: 8017474
• 负责人: JULIE L KADRMAS
• 金额: $ 28.03万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8017474
• 关键词: Actins; Address; Adhesions; Affect; Affinity; Binding; Binding Proteins; Biological Process; CD47 Antigen; Cell Adhesion; Cell Nucleus; Cell membrane; Cell-Cell Adhesion; Cells; Cellular Immunity; Complement; Complex; Coupled; Coupling; Cultured Cells; Cytoplasmic Tail; Cytosol; Data; Defect; Deposition; Disease; Disseminated Malignant Neoplasm; Drosophila genus; Embryo; Embryonic Development; Event; Exhibits; Extracellular Matrix; Gene Expression; Goals; Human; Human Pathology; Immune response; Inflammatory Response; Integrins; LHX1 gene; LIM Domain; LIMS1 gene; Learning; Leucine-Rich Repeat; Location; Malignant - descriptor; Malignant Neoplasms; Mediating; Membrane; Molecular; Molecular Mechanisms of Action; Neoplasm Metastasis; Neurons; Nuclear; Nuclear Export; Nuclear Translocation; Oncogenic; Phenotype; Phosphotransferases; Play; Preventive Intervention; Process; Protein Binding; Protein Binding Domain; Proteins; Recruitment Activity; Regulation; Regulatory Pathway; Relative (related person); Research; Role; Scaffolding Protein; Series; Signal Pathway; Signal Transduction; Site; Source; Stimulus; Structure; Therapeutic Intervention; Tissues; Tumor Cell Invasion; Work; Wound Healing; cell motility; cohort; genetic regulatory protein; insight; integrin-linked kinase; interest; leucine-rich repeat protein; migration; mutant; novel; null mutation; public health relevance; ras Oncogene; receptor; research study; scaffold; therapeutic target; tumor; tumor progression

DESCRIPTION (provided by applicant): Regulated cell adhesion and migration are critical for a variety of biological processes including early embryonic development, neuronal path-finding, inflammatory response, wound healing and tissue integrity, cell-mediated immunity, and tumor invasion and metastasis. Integrin trans-membrane receptors are key regulators of cell migration, and act through an intricate assembly of regulatory proteins associated with the integrin cytoplasmic domain, many of which function as molecular scaffolds. The goal of this research is to identify novel regulatory pathways by which scaffolding proteins modulate cell adhesion and migration via regulated binding of protein partners. In previous work, we characterized the LIM domain scaffold PINCH, which is required for integrin-dependent cell adhesion, migration and signaling. PINCH binds with high affinity to both Integrin-linked kinase (ILK) and Ras suppressor 1 (RSU1), a Leucine-rich repeat protein that suppresses transformation of cultured cells expressing activated Ras, an oncogene present in 30% of all human tumors. We propose that the PINCH scaffold serves as a node for modulating integrin signaling by regulated recruitment of partners like ILK and RSU1, coupled to other signaling pathways. Through a series of experiments in Drosophila in which we specifically disrupt binding interactions with PINCH and alter the subcellular localization of components of the PINCH complex, we will dissect the requirement for ILK and RSU1 binding in carrying out the integrin-dependent functions of PINCH. Our preliminary data indicates that binding to ILK localizes PINCH within integrin-associated complexes at the cell membrane, and regulates the shuttling of PINCH to the nucleus. We will determine if PINCH has essential functions in the nucleus, and whether it has an overlapping complement of binding partners in the cytosol versus the nucleus. Additional data suggest that RSU1 is sequestered in integrin-associated complexes by its interaction with PINCH. We will examine how PINCH-RSU1 binding is regulated, and identify partners and activities for RSU1 that are independent of PINCH. A molecular understanding of how PINCH regulates cell adhesion and migration through its scaffolding activity may ultimately suggest strategies for prevention and therapeutic intervention in human pathologies like metastatic cancer, in which aberrant cell migration plays a prominent role. Public Health Relevance: Cell adhesion and migration are critical components of many biological processes including embryonic development, inflammatory and immune response, wound healing, and tumor invasion and metastasis. Cell adhesion and migration are controlled by an intricate network of regulatory circuits. This research will define new molecular mechanisms by which protein interactions regulate adhesion and migration, with the ultimate goal of identifying potential therapeutic targets for diseases arising from aberrant cell migration, such as the malignant progression of tumors.

描述(申请人提供)：受调控的细胞黏附和迁移对多种生物学过程至关重要，包括早期胚胎发育、神经元路径寻找、炎症反应、伤口愈合和组织完整性、细胞介导的免疫以及肿瘤的侵袭和转移。整合素跨膜受体是细胞迁移的关键调节因子，通过与整合素细胞质结构域相关的调控蛋白的复杂组装发挥作用，其中许多蛋白起到分子支架的作用。本研究的目的是确定新的调控途径，支架蛋白通过调节蛋白伙伴的结合来调节细胞的黏附和迁移。在以前的工作中，我们表征了LIM结构域支架夹点，这是整合素依赖的细胞黏附、迁移和信号传递所必需的。PINCH与整合素连接的激酶(ILK)和RAS抑制因子1(RSU1)具有高亲和力，RSU1是一种富含亮氨酸的重复蛋白，可以抑制表达活化RAS的培养细胞的转化，RAS是一种存在于30%的人类肿瘤中的癌基因。我们认为，Pinch支架作为一个节点，通过调节ILK和RSU1等伙伴的招募来调节整合素信号，并与其他信号通路相连。通过在果蝇身上进行的一系列实验，我们特异性地干扰了与Pinch的结合作用，并改变了Pinch复合体成分的亚细胞定位，我们将剖析在实现Pinch的整合素依赖功能时对ILK和RSU1结合的要求。我们的初步数据表明，与ILK的结合使Pinch定位于细胞膜上的整合素相关复合体，并调节Pinch的穿梭到细胞核。我们将确定Pinch是否在细胞核中具有必要的功能，以及它是否在胞浆中与细胞核中有重叠的结合伙伴互补。更多的数据表明，RSU1通过与Pinch的相互作用隔离在整合素相关的复合体中。我们将研究Pinch-RSU1结合是如何调控的，并确定RSU1的伙伴和活动独立于Pinch。对Pinch如何通过其支架活动调节细胞黏附和迁移的分子理解最终可能会为预防和治疗人类病理疾病(如转移性癌症)提供策略，在这些疾病中，异常的细胞迁移起着突出的作用。公共卫生相关性：细胞黏附和迁移是许多生物学过程的关键组成部分，包括胚胎发育、炎症和免疫反应、伤口愈合以及肿瘤侵袭和转移。细胞黏附和迁移由一个错综复杂的调节电路网络控制。这项研究将定义蛋白质相互作用调节黏附和迁移的新分子机制，最终目标是确定由异常细胞迁移引起的疾病的潜在治疗靶点，如肿瘤的恶性进展。