Activation of Aurora Kinase A by DNA damage - understanding the signalling pathway to improve radiotherapy response

放射治疗是癌症治疗的主要手段，然而放射抵抗是治疗局部晚期、复发和转移性癌症的一个重大挑战。迫切需要新的方法来改善难治性癌症的放射反应。我们发现，临床上丝氨酸/苏氨酸激酶Aurora A的抑制剂(AURKA)是癌细胞的放射增敏剂，并且AURKA的过度表达与包括肺癌在内的癌症的放射抵抗有关。有趣的是，AURKA在电离辐射诱导DNA损伤后以延迟动力学激活。这一令人兴奋的研究方向主要集中在鉴定和表征DNA损伤(包括电离辐射)激活的功能相关的AURKA依赖信号通路的磷酸蛋白质组学方法。该项目将为AURKA在DNA损伤后的作用提供新的机械性见解，并最终补充和指导未来的临床前研究。具体目标：(1)辐射诱导DNA损伤后DNA损伤反应和(磷酸)蛋白质组的定量蛋白质组分析。(2)DNA损伤后AURKA的激活和失活及其下游靶点激活的途径和靶点分析以及(3)与肿瘤放射治疗相关的AURKA信号网络的体内分析。实验方法：在临床前药物开发涉及的广泛技能方面，包括SILAC和基于TMT的质谱学生化和细胞生物学分析(如DNA损伤、细胞周期和体外激酶分析)，以及对AURKA及其下游靶点的体内分析(非小细胞肺癌(NSCLC)标本)。环境：在谢菲尔德，学生将成为Bryant实验室的一部分，一个支持性的，充满活力的多学科团队，从化学家到临床研究员，他们都在共同努力，开发更好的癌症治疗方法。特别是，这名学生将与一名临床研究员并行工作，后者目前正在评估AURKA抑制的临床意义，并开发放射增强剂的临床试验。作为谢菲尔德核酸研究所和谢菲尔德ECMC中心的一部分，您还将受益于谢菲尔德更广泛的研究。这名学生还将在利物浦大学蛋白质组研究中心的艾尔斯实验室接受培训，该中心是利物浦大学最先进的质谱学设施。在这里，这名学生将加入一个充满活力的生物化学家团队，参与开发分析信号通路的新技术。

"Radiation is a mainstay of cancer therapy, however radioresistance is a significant challenge in the treatment of locally advanced, recurrent and metastatic cancers. Novel ways to improve radiation response in hard to treat cancers are urgently needed.We find that clinical inhibitors of the serine/threonine kinase Aurora A (AURKA) are cancer cell radiosensitizers, and that AURKA overexpression correlates with radioresistance in cancer, including lung cancer. Interestingly, AURKA is activated with delayed kinetics following DNA damage induced by ionizing radiation. This exciting studentship focuses on phosphoproteomic approaches for identification and characterization of functionally relevant AURKA-dependent signaling pathways activated by DNA damage (including ionizing radiation). The project will provide new, mechanistic insights into the role of AURKA following DNA damage, and will ultimately complement and guide future pre-clinical studies.Specific objectives: (1) Quantitative proteomic analysis of the DNA damage response and (phospho)proteome following Radiation-induced DNA damage. (2) Pathway and target analysis of AURKA activation and inactivation and downstream target activation after DNA damage and (3) In vivo analysis of AURKA signaling networks relevant to tumour radiotherapy.Experimental Approach: The studentship offers a unique training opportunity in a broad range of skills involved in pre-clinical drug development, including SILAC and TMT-based Mass spectroscopy biochemical and cell biology assays (such as DNA damage, cell cycle and in vitro kinase assays) and in vivo analysis of AURKA and its downstream targets in patient material (Non Small Cell Lung Cancer (NSCLC) specimens).Environment: In Sheffield the student will be part of the Bryant lab, a supportive, dynamic multidisciplinary team spanning chemists to clinical fellows, who are all working together to develop better treatments for cancer. In particular the student will work in parallel to a clinical fellow who is currently evaluating the clinical implications of AURKA inhibition and developing clinical trails of radiosenstizing agents. You will also benefit from being part of the wider research within Sheffield as part of the Sheffield Institute for Nucleic Acids and Sheffield ECMC centers. The student will also spend time training in the Eyers lab in the Centre for Proteome Research, a state of the art mass-spectrometry facility at University of Liverpool. Here the student will join a vibrant group of biochemists involved in developing novel techniques for analyzing signaling pathways."