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

项目摘要 乳腺肿瘤通常根据其与正常乳腺组织相比的表观硬度来识别， 乳腺癌患者组织硬度的增加通常与转移的增加相关。当 人乳腺上皮细胞在具有乳腺肿瘤基质硬度的3D培养物中生长， 更恶性的表型组织硬度增加与肿瘤浸润和转移之间的联系 分子水平还不太清楚。杨实验室先前的研究表明， 因子Twist 1通过其诱导上皮-间质转化的能力是转移的关键调节因子 (EMT)这是一种发育程序，也被癌细胞用来入侵和转移。 Yang实验室发现，随着基质硬度的增加，Twist 1从细胞质易位到细胞质中， 原子核在低基质硬度下，Twist 1与其细胞质锚GT3活化蛋白结合 结合蛋白2（G3 BP 2）。这种相互作用由Twist 1上的酪氨酸107（Y107）的磷酸化控制， 其又从G3 BP 2释放Twist 1进入细胞核。初步研究以确定激酶负责 对于这种磷酸化事件，鉴定了一种酪氨酸激酶，其导致Twist 1在低基质中的核积累， 刚度，这表明该候选激酶是必需的Twist 1在低刚度的细胞质定位。 因此，我假设这种候选激酶控制Twist 1的亚细胞定位， 通过Twist 1机械调节途径改变基质硬度。为了验证这个假设，我计划1-2） 确定该候选人在该机械传导途径中发挥的作用; 3）确定该候选人在该机械传导途径中发挥的作用 Twist 1依赖的体内侵袭和转移中的候选激酶。

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.