Characterization and functional analysis of the new microtubule associated protein RITA in the development of breast cancer

新型微管相关蛋白RITA在乳腺癌发生过程中的表征及功能分析

• 批准号: 508177751
• 负责人: Professorin Dr. Juping Yuan
• 金额: --
• 依托单位: Klinik für Frauenheilkunde und Geburtshilfe
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/508177751?language=en
• 关键词: Characterization; functional; analysis; new; microtubule

Breast cancer, a global health concern, is the primary cause of death among women worldwide. Despite significant achievements, breast cancer metastasis remains largely incurable. Uncontrolled cell migration and invasion is central to the metastatic process. Microtubules exert spatiotemporal control in cell migration and invasion. Being associated with various regulatory and motor proteins, they interact with diverse components of the cell migration machinery and regulate cell polarity, intracellular trafficking and focal adhesion turnover. Dysfunctional microtubules result in uncontrolled migration of malignant cells and promote the development of malignancy. We have identified that RITA, the RBP-J interacting and tubulin associated protein, is crucial in regulating microtubule dynamics and the activation of Aurora A, a frequently deregulated kinase in various tumors, suggesting the involvement of RITA in oncogenesis. In fact, RITA is highly expressed in anal carcinoma tissues and its expression level is negatively correlated with prognosis and therapy response of patients. Recently, we report that RITA is tightly correlated with migration/invasion of breast cancer cells by affecting the turnover of focal adhesions and dynamics of the cytoskeleton. The precise molecular mechanisms remain however undefined. Interestingly, RITA is associated with the lipoma preferred partner (LPP), an important player in migration and invasion of breast cancer cells. With this proposal, we will investigate how RITA affects the motility of breast cancer cells and its significance in breast cancer development. In particular, using CRISPR/Cas9 RITA knockout/in cell lines, RITA knockdown/knockout mouse fibroblasts, fluorescence recovery after photo-bleaching, living cell imaging, transcriptomic analysis, primary breast cancer tissues as well as other molecular/biochemical/cellular methodology, we would like to address following issues: 1. The molecular mechanisms by which RITA affects migration and invasion of breast cancer cells 2. The expression levels of RITA and LPP in primary breast cancer tissues. 3. Correlation of the RITA expression with tumor microenvironment and clinical parameters of breast cancer patients. Given the crucial roles of RITA in regulating microtubule dynamics and its involvement in oncogenesis, this study will provide the molecular mechanisms by which deregulated RITA affects breast cancer development, therapy response and prognosis of breast cancer patients.

乳腺癌是一个全球性的健康问题，是全世界妇女死亡的主要原因。尽管取得了重大成就，但乳腺癌转移在很大程度上仍然无法治愈。不受控制的细胞迁移和侵袭是转移过程的核心。微管在细胞迁移和侵袭中发挥时空控制作用。与各种调节蛋白和马达蛋白相关，它们与细胞迁移机制的各种组分相互作用，并调节细胞极性、细胞内运输和粘着斑周转。微管功能障碍导致恶性细胞不受控制的迁移并促进恶性肿瘤的发展。我们已经确定，RITA，RBP-J相互作用和微管蛋白相关蛋白，是至关重要的调节微管动力学和激活极光A，在各种肿瘤中经常失调激酶，这表明RITA参与肿瘤发生。事实上，RITA在肛门癌组织中高度表达，其表达水平与患者的预后和治疗反应呈负相关。最近，我们报告说，RITA是密切相关的乳腺癌细胞的迁移/侵袭，通过影响营业额的局灶性粘连和动态的细胞骨架。然而，精确的分子机制仍然不确定。有趣的是，RITA与脂肪瘤首选伴侣（LPP）相关，LPP是乳腺癌细胞迁移和侵袭的重要参与者。本研究将探讨RITA如何影响乳腺癌细胞的运动性及其在乳腺癌发展中的意义。特别地，使用CRISPR/Cas9 RITA敲除/在细胞系中，RITA敲除/敲除小鼠成纤维细胞，光漂白后的荧光恢复，活细胞成像，转录组学分析，原发性乳腺癌组织以及其他分子/生物化学/细胞方法，我们想要解决以下问题：RITA影响乳腺癌细胞迁移和侵袭的分子机制2. RITA和LPP在乳腺癌组织中的表达水平3.乳腺癌患者RITA表达与肿瘤微环境及临床参数的相关性研究鉴于RITA在调节微管动力学和参与肿瘤发生中的关键作用，本研究将提供RITA失调影响乳腺癌发展、治疗反应和乳腺癌患者预后的分子机制。