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The role of PKD2 in HSP90,- hypoxia- and TGF beta-mediated epithelial-to-mesenchymal transition and metastasis

PKD2 在 HSP90、缺氧和 TGFβ 介导的上皮间质转化和转移中的作用

基本信息

批准号：
232091324
负责人：
Dr. Ninel Azoitei
金额：
--
依托单位：
Klinik für Innere Medizin I
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2013
资助国家：
德国
起止时间：
2012-12-31 至 2018-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/232091324?language=en
关键词：
role PKD2 HSP90 hypoxia TGF

项目摘要

The proposal focuses on the investigation of protein kinase D2s contribution to epithelial-to-mesenchymal transition (EMT), a process implicated in tumor metastasis and initiated by multiple factors such as hypoxia, transcription factors or growth factors. The investigations detailed in the second proposal of the running grant will engage meticulous cellular models dedicated to studying EMT. Detailed morphologic analysis of tumor cells cultivated on porous support and/or 3D collagen gels will be corroborated with various biochemical, immunofluorescence and immunohistochemical analysis. The in vitro experiments will be substantiated by human tumor xenograft on CAM as well as analysis of EMT protein markers on tumors xenografted in nude mice, already available from the previous project. Furthermore, the in vivo contribution of PKD2 to the metastatic potential of mammary cancer cells will be delineated in new experiments with immunodeficient athymic mice. Investigations assessing the role of PKD2 during EMT will contribute to the broadening of our knowledge with respect to the requirement of kinase for other aspects of tumorigenesis, unknown so far for this kinase. The proposed work program will bring insight to participation of PKD2 to EMT initiated and maintained by TGF-beta, HSP90 chaperone and hypoxia. Answering whether PKD2 regulates EMT via the same molecular mechanism/s that govern tumor angiogenesis (e.g. via HIF-1 alpha), or whether the kinase is implicated in other potential signaling pathways that favor the transition of epithelial cancer cells to a mesenchymal phenotype will represent a central focus of the extension proposal. Several microarrays assessing PKD2 involvement in the regulation of several crucial molecules (targeted by NF- kappa B, but not only) reported to emanate signals to induce EMT as well as the planned Mass spectrometry-based interactome on hypoxic cancer cells that undergo EMT will not only serve the current work program, but will also help investigations beyond this proposal. Finally, the use of HSP90-inhibitor PU-H71 (currently in clinical development) will promote our findings for a conceivable use in an immediate therapeutic approach targeting PKD2 stability in particularly hypoxic human tumors.

该提案侧重于研究蛋白激酶D2s在上皮向间充质转化(EMT)中的作用，EMT是一种与肿瘤转移有关的过程，由缺氧、转录因子或生长因子等多种因素启动。在运行拨款的第二个提案中详细说明的调查将使用致力于研究EMT的细致的细胞模型。在多孔载体和/或3D胶原凝胶上培养的肿瘤细胞的详细形态分析将得到各种生化、免疫荧光和免疫组织化学分析的证实。体外实验将通过将人肿瘤移植到CAM上以及对裸鼠移植瘤的EMT蛋白标记物的分析来证实，这一分析已经从先前的项目中获得。此外，PKD2在体内对乳腺癌细胞转移潜能的贡献将在新的免疫缺陷裸鼠实验中描绘出来。评估PKD2在EMT中的作用的研究将有助于拓宽我们对该激酶在肿瘤发生的其他方面所需的知识，这一点到目前为止还不清楚。拟议的工作计划将为PKD2参与由转化生长因子-β、HSP90伴侣和低氧启动和维持的EMT带来深刻的认识。回答PKD2是否通过调控肿瘤血管生成的相同分子机制(例如，通过HIF-1α)调节EMT，或者该激酶是否涉及有利于上皮癌细胞向间充质表型转变的其他潜在信号通路，将是扩展提案的中心焦点。几个评估PKD2参与调节几个关键分子(以核因子-kappa B为靶标，但不仅是如此)的微阵列被报告发出信号以诱导EMT，以及计划中的对接受EMT的缺氧癌细胞进行基于质谱学的相互作用组，这不仅将服务于当前的工作计划，而且还将有助于超出这一提议的调查。最后，HSP90抑制剂PU-H71(目前在临床开发中)的使用将促进我们的发现，可以想象在针对PKD2稳定性的直接治疗方法中使用，特别是在缺氧的人类肿瘤中。