去甲基化酶ALKBH5表观调控TGIF-1/LDHA轴激活糖酵解途径介导胶质瘤替莫唑胺耐药的分子机制研究

替莫唑胺（TMZ）耐药是制约胶质瘤患者术后疗效的主要原因，申请人前期报道去甲基化酶ALKBH5高表达与TMZ耐药和患者预后不良相关，但其机制尚未阐明。近期应用m6A-MeRIP-seq和RIP技术确定ALKBH5可能通过m6A修饰调控TGIF-1表达；利用生信分析和体外实验初步证实TGIF-1通过激活糖酵解途径关键酶LDHA，增加胶质瘤细胞对TMZ的抗药性。我们由此提出本项目科学假说：ALKBH5表观调控TGIF-1/LDHA轴激活糖酵解途径介导胶质瘤TMZ耐药。本课题拟利用耐药细胞模型、动物模型及胶质瘤患者组织标本，证实ALKBH5通过TGIF-1调控胶质瘤TMZ耐药；结合MeRIP-seq、Co-IP等技术，进一步明确ALKBH5通过m6A修饰调控TGIF-1/LDHA轴，激活糖酵解途径介导胶质瘤TMZ耐药的机制。通过本项目研究，为阐明TMZ耐药机制及开展逆转耐药研究提供新的科学依据。

Temozolomide (TMZ) resistance is the main reason restricting the postoperative efficacy of glioma patients, the applicant previously reported that the high expression level of demethylase ALKBH5 was associated with TMZ resistance and poor prognosis of patients, but the mechanism has not been clarified yet. Recently, m6A-MeRIP-seq and RIP techniques were used to determine that ALKBH5 may regulate TGIF-1 expression through m6A methylation modification. Bioinformatics analysis and in vitro experiments preliminary confirmed that TGIF-1 increased the resistance of glioma cells to TMZ by activating LDHA, the key enzyme in the glycolytic pathway. Therefore, we proposed the scientific hypothesis of this project: ALKBH5 activates the glycolytic pathway to mediate TMZ resistance in gliomas via epigenetic regulation of the TGIF-1/LDHA axis. In this study, TMZ resistant cell models, animal models and tissue specimens of glioma patients were used to confirm that ALKBH5 regulates drug resistance of TMZ through TGIF-1. The mechanism of ALKBH5 activates glycolysis pathway to mediate temozolomide resistance in gliomas via m6A methylation modification regulation of TGIF/LDHA axis was further clarified combined with MeRIP-seq, Co-IP and other technologies. The research of this project provides a new scientific basis for elucidating the mechanism of TMZ resistance and carrying out reverse drug resistance research.