Talin2-mediated traction force drives MMP secretion and cell invasion

Talin2介导的牵引力驱动MMP分泌和细胞侵袭

• 批准号: 9397621
• 负责人: CAI HUANG
• 金额: $ 30.03万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-30 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9397621
• 关键词: Ablation; Actins; Address; Affinity; Antibodies; Automobile Driving; Binding; Biochemical; Cell Count; Cell membrane; Cell physiology; Cells; Cellular biology; Complex; Computational Technique; Confocal Microscopy; Congenital Camptodactyly; Cytoskeleton; Data; Development; ERBB2 gene; Enzymes; Fingers; Focal Adhesions; Generations; Genes; Goals; Golgi Apparatus; Hereditary Disease; Human Genetics; Imaging Techniques; Integrin beta Chains; Integrins; Interstitial Collagenase; Leucine; MMP9 gene; Matrix Metalloproteinases; Mediating; Methodology; Microscopy; Modeling; Molecular; Mutation; Myosin ATPase; Neoplasm Metastasis; Nonmuscle Myosin Type IIA; Pharmaceutical Preparations; Phosphatidylinositol 4,5-Diphosphate; Phosphatidylinositols; Phosphotransferases; Positioning Attribute; Production; Proteins; Research; Resolution; Role; Tail; Talin; Testing; Traction; Trastuzumab; Vesicle; Work; base; cancer cell; cancer therapy; cell motility; design; experience; extracellular; genome editing; link protein; malignant breast neoplasm; non-muscle myosin; novel; phosphatidylinositol 4-phosphate; protein complex; targeted treatment; therapy design; trafficking

ABSTRACT Talin is a key component in the intracellular complex of proteins linking the extracellular martrix to the actin cytoskeleton via integrins at the plasma membrane. Talin interacts with β-integrin tails and actin to control integrin activation and force generation. It also binds to phosphatidylinositol 4 phosphate 5-kinase type I γ 90 (PIPKIγ90), an enzyme that catalyzes the production of phosphatidylinositol 4,5-bisphosphate (PIP2), and regulates vesicle trafficking. In this capacity, talin regulates focal adhesion dynamics, force generation and invadopodium stability, thus governing cell migration and invasion. There are two talin genes, Tln1 and Tln2, which encode talin1 and talin2; talin1 has been well studied and it is generally believed that talin2 functions redundantly with talin1. However, our preliminary data reveal some important differences between talin 1 and 2. Specifically, talin2 has stronger binding to β integrin tails than talin1 and Ser339 of talin2 is largely responsible for this affinity difference. Intriguingly, a new study shows that Fifth Finger Camptodactyly, a human genetic disease, is caused by a Leu mutation at talin2 Ser339, suggesting that the talin2-integrin interaction may be important for development. Talin2 is localized at invadopodia and is indispensable for traction force generation and invadopodium maturation. And, ablation of talin2 but not talin1 abolished the secretion of matrix metalloproteinase (MMP) 1 and 9. In this regard, talin2 promotes PIPKIγ90 binding to Exo84 (a component of the exocyst involved in vesicle secretion) and interacts with non-muscle myosin IIA (NMMIIA), a major myosin involved in traction force generation. Based on these data, our central hypothesis is that talin2 interacts with NMMIIA to mediate traction force generation and promotes PIPKIγ90-Exo84 complex formation, thus driving MMP1 and MMP9 secretion, invadopodium maturation and cell invasion. We will test this hypothesis by addressing the following SPECIFIC AIMS: 1) define how talin2 regulates MMP secretion, invadopodium maturation and cell invasion; 2) dissect how talin2 modulates PIPKIγ90-Exo84 interaction to regulate MMP secretion, invadopodium maturation and cell invasion; 3) determine how talin2 interacts with NMMIIA to regulate traction force generation, MMP secretion, invadopodium maturation and cell invasion. Our proposed studies are designed to uncover a novel, fundamental role for talin2 in regulating MMP secretion, invadopodium maturation and cancer cell invasion and the underlying mechanisms. Support for this connection is provided by the fact that talin2 was found to be down-regulated by Trastuzumab, a HER2-targeting antibody drug for breast cancers and our findings indicate that talin2 is essential to the traction force development required for invadopodium maturation and cell invasion. Thus, talin2 could be a target for therapies designed to inhibit the first steps of metastasis.

摘要 塔林蛋白是连接细胞外基质和肌动蛋白的细胞内蛋白复合物的关键成分 细胞骨架通过整合素在质膜上。Talin与β-整联蛋白尾部和肌动蛋白相互作用以控制整联蛋白 激活和力生成。它还结合磷脂酰肌醇4磷酸5-激酶I γ 90（PIPKIγ90）， 一种催化磷脂酰肌醇4，5-二磷酸（PIP 2）产生并调节囊泡的酶 贩卖人口在这种能力下，talin调节粘着斑动力学，力的产生和侵袭足的稳定性， 从而控制细胞迁移和侵入。存在两个talin基因，Tln 1和Tln 2，其编码talin 1， Talin 2; Talin 1已经被充分研究，并且通常认为Talin 2与Talin 1具有冗余的功能。 然而，我们的初步数据揭示了talin 1和2之间的一些重要差异。具体来说，塔林2号 与β整联蛋白尾的结合比talin 1和talin 2的Ser 339更强，这主要是这种亲和力差异的原因。 有趣的是，一项新的研究表明，五指弯曲症，一种人类遗传疾病，是由一种亮氨酸引起的。 talin 2 Ser 339突变，表明talin 2-整合素相互作用可能对发育很重要。 Talin 2定位于内陷伪足，并且对于牵引力的产生和内陷伪足是不可缺少的 成熟并且，切除talin 2而不是talin 1可抑制基质金属蛋白酶1（MMP 1）的分泌 和9.在这方面，talin 2促进PIPKIγ90与Exo 84（参与囊泡形成的外囊的组分）结合， 分泌），并与非肌肉肌球蛋白IIA（NMMIIA），一种主要的肌球蛋白参与牵引力 一代基于这些数据，我们的中心假设是talin 2与NMMIIA相互作用以介导牵引 力产生并促进PIPKIγ90-Exo 84复合物形成，从而驱动MMP 1和MMP 9分泌， 侵入柄成熟和细胞侵入。我们将通过解决以下具体问题来检验这一假设。 目的：1）确定talin 2如何调节MMP分泌，侵袭足成熟和细胞侵袭; 2）分析talin 2如何调节MMP分泌，侵袭足成熟和细胞侵袭。 talin 2通过调节PIPKIγ90-Exo 84相互作用调节MMP分泌、侵袭足成熟和细胞增殖 侵袭; 3）确定talin 2如何与NMMIIA相互作用以调节牵引力产生，MMP分泌， 侵入柄成熟和细胞侵入。我们提出的研究旨在揭示一种新的、基本的 talin 2在调节MMP分泌、侵袭足成熟和癌细胞侵袭中的作用及其潜在的 机制等支持这种联系的事实是，talin 2被发现下调， 曲妥珠单抗，一种针对乳腺癌的HER 2靶向抗体药物，我们的研究结果表明talin 2是必不可少的 到侵袭足成熟和细胞侵入所需的牵引力发展。因此，talin 2可以是 一种旨在抑制转移第一步的治疗靶点。