The role of Tyk2 in Twist1-mediated EMT and metastasis in response to matrix stiffness

Tyk2 在 Twist1 介导的 EMT 和响应基质硬度的转移中的作用

• 批准号: 9258357
• 负责人: Hannah E Majeski
• 金额: $ 3.59万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9258357
• 关键词: Asses; Binding; Binding Proteins; Biochemical; Breast Cancer Patient; Breast Cancer cell line; Breast Epithelial Cells; Cell Nucleus; Cytoplasm; Data; Development; Distant Metastasis; Embryonic Development; Epithelial; Epithelial Cells; Event; GTPase-Activating Proteins; Goals; Hardness; Human; Hydrogels; Invaded; Link; Mammary Gland Parenchyma; Mammary Neoplasms; Mammary gland; Mediating; Mesenchymal; Molecular; Morphology; Mus; Neoplasm Metastasis; Nuclear; Pathway interactions; Phosphorylation; Phosphorylation Site; Phosphotransferases; Play; Protein Dephosphorylation; Protein Tyrosine Kinase; Regulatory Pathway; Research; Role; Signal Transduction; System; TYK2; Testing; Tissues; Translating; Tumor Cell Invasion; Tyrosine Phosphorylation; Work; Xenograft Model; Yang; base; cancer cell; clinically significant; in vivo; knock-down; malignant breast neoplasm; malignant phenotype; mechanical force; mechanotransduction; mutant; neoplastic cell; novel; outcome forecast; programs; promoter; response; three dimensional cell culture; transcription factor; tumor; tumor microenvironment; tumor xenograft

Project Summary Breast tumors are often identified based on their apparent hardness compared to normal breast tissue, and in breast cancer patients an increase in tissue rigidity is often correlated with an increase in metastasis. When human mammary epithelial cells are grown in 3D culture with the matrix stiffness of breast tumors, they develop a more malignant phenotype. The link between increased tissue rigidity and invasion and metastasis at the molecular level is not well understood. Previous research from the Yang lab has shown that the transcription factor Twist1 is a key regulator of metastasis through its ability to induce Epithelial-Mesenchymal Transition (EMT), a developmental program also used by cancer cells to invade and metastasize. The Yang lab discovered that upon an increase in matrix stiffness, Twist1 translocates from the cytoplasm into the nucleus. Under low matrix stiffness, Twist1 is bound to its cytoplasmic anchor GTPase activating protein binding protein 2 (G3BP2). This interaction is controlled by phosphorylation of tyrosine 107 on Twist1 (Y107), which in turn releases Twist1 from G3BP2 to enter the nucleus. Initial studies to identify the kinase responsible for this phosphorylation event identified a tyrosine kinase that caused Twist1 nuclear accumulation at low matrix stiffness, suggesting that this candidate kinase is required for cytoplasmic localization of Twist1 at low stiffness. Therefore, I hypothesize that this candidate kinase controls Twist1 subcellular localization in response to changes in matrix stiffness via the Twist1 mechanoregulation pathway. To test this hypothesis I plan to 1-2) Determine the role that this candidate plays in this mechanotransduction pathway; 3) determine the role of this candidate kinase in Twist1 dependent invasion and metastasis in vivo.

项目摘要 乳腺肿瘤通常是根据与正常乳腺组织相比的表观硬度来识别的，而且在 乳腺癌患者组织硬度的增加往往与转移的增加有关。什么时候 人乳腺上皮细胞在3D培养中生长，具有乳腺肿瘤的基质硬度，它们会发育 一种更恶性的表型。组织刚性增加与肿瘤的侵袭和转移之间的联系 分子水平还没有被很好地理解。杨实验室之前的研究表明，转录 Twist1因子诱导上皮间充质转化是肿瘤转移的关键调节因子 (EMT)，一种发育程序，也被癌细胞用来侵袭和转移。 杨的实验室发现，随着基质硬度的增加，Twist1从细胞质转移到 原子核。在低基质刚性条件下，Twist1与其胞质锚GTP酶激活蛋白结合 结合蛋白2(G3BP2)。这种相互作用由Twist1(Y107)上酪氨酸107的磷酸化控制， 进而从G3BP2释放Twist1进入原子核。初步研究以确定相关的激酶 对于这种磷酸化事件，确定了一种酪氨酸激酶，它导致Twist1核在低基质处聚集 低硬度时，Twist1的胞质定位需要该候选蛋白。 因此，我推测该候选激酶控制Twist1亚细胞定位，以响应 通过Twist1机械调节途径改变基质硬度。为了检验这一假设，我计划使用1-2) 确定该候选者在该机械转导途径中所扮演的角色；3)确定这一角色 Twist1依赖于体内侵袭和转移的候选蛋白激酶。