EIF5A2通过TAZ维持膀胱癌细胞干性及促进肿瘤吡柔比星化疗耐药的机制研究

Our previous study found that the recurrence rate of non muscle invasive bladder cancer in patients with EIF5A2 high expression was significantly higher than that of EIF5A2 low expression after application of pirarubicin intravesical instillation treatment. Our in vitro experiments showed that the sensitivity to pirarubicin in EIF5A2 overexpression bladder cancer cell line was significantly lower than the control cell line, but the molecular mechanism is not clear. In this project, we intend to firstly detect the sensitivity to pirarubicin, the ability to self renew and the expression of cancer stemness markers in EIF5A2 overexpression/knockdown bladder cancer cells by in vitro and in vivo experiments, to confirm the role of EIF5A2 in bladder cancer cells stemness and chemotherapeutic resistant. Secondly, we intend to use immunoprecipitation, point-mutation, and sequence deletion technologies to explore the mechanisms of EIF5A2-PRPs-TAZ signal axis in the regulations of bladder cancer cells stemness and pirarubicin resistant. Finally, the effect of nanoparticles delivering small interfering RNA targeting EIF5A2 combined with pirarubicin intravesical instillation in the treatment of bladder cancer is intended to be tested. We hope to elucidate the molecular mechanism of EIF5A2 in promoting pirarubicin resistance of bladder cancer cells, and to provide the experimental basis for further understanding the molecular mechanisms of bladder cancer progression and the selection of therapeutic targets.

我们前期研究发现非肌层浸润性膀胱癌术后应用吡柔比星膀胱腔内灌注患者中EIF5A2高表达组的复发率显著高于EIF5A2低表达组，体外实验发现EIF5A2过表达膀胱癌细胞株对吡柔比星药物敏感性显著差于对照细胞株，但其分子机制尚不明确。本项目拟首先检测EIF5A2基因过表达和敲除后膀胱癌细胞株体内外对化疗药物吡柔比星敏感性、自我更新能力及干性相关标记物表达水平的变化，明确EIF5A2对膀胱癌细胞化疗药物治疗耐药及干性的影响；其次，通过免疫共沉淀及点突变序列缺失等实验探索EIF5A2-PRPs-TAZ信号轴在膀胱癌细胞干性调控及吡柔比星耐药中的机制。最后，通过大鼠原位膀胱癌模型观察纳米材料携带小干扰RNA靶向EIF5A2联合吡柔比星腔内灌注治疗膀胱癌的疗效。期望阐明EIF5A2促进膀胱癌细胞吡柔比星耐药的分子调控机制，为深入理解膀胱癌恶性进展的分子机制及治疗靶点的选择提供实验基础。

目前肿瘤辅助治疗的临床应用中缺乏有效的指标指导治疗。针对这一问题，申请人专注于肿瘤转移复发与耐药的预测分子基础与临床转化研究。①：膀胱肿瘤分子分型与精准治疗的转化医学研究，建立m6A免疫相关lncRNA模型能够预测膀胱癌患者的生存预后、免疫微环境状态以及化疗敏感程度，探索TP53/PIK3CA/ATM轴突变、免疫表型差异对膀胱癌免疫治疗效果的影响，对于于膀胱癌的诊断和个体化治疗具有重大意义(J Transl Med,Front Immunol,Front Cell Dev Biol);②：阐明EIF5A2在膀胱癌发生发展与腔内复发、前列腺癌发生发展中的重要作用( Cancer Manag Res, Future Oncol, Exp Ther Med)，探索EIF5A2 可以通过激活 TAZ 维持膀胱肿瘤细胞干性并促进非肌层浸润性膀胱癌术后膀胱内灌注化疗耐药机制，同时以小干扰 RAN 靶向 EIF5A2 与纳米化表柔比星联用可以显著改善膀胱肿瘤的进展、复发与 化疗耐药。③：通过高通量筛选泌尿系肿瘤中分子标记物，以SNP构建分类器用于预测肾细胞癌局部复发的风险，改善术后辅助治疗方案的决策(Lancet Oncol)，SYNE1突变、ZDHHC1、肿瘤自噬相关lncRNA对于肿瘤发生发展、复发与转移、免疫微环境重塑的影响(aging-us, Oxid Med Cell Longev,Gnomecs);近五年发表(共同)通讯/第一作者论著13篇，8篇IF>5,单篇最高IF26.5.在项目实行期间，项目第一完成人陈炜教授在项目完成期间获聘二级教授，项目主要成员张家兴获教育部青年长江学者。按期完成项目预期的所有研究目标