YOD1调控胶质瘤细胞糖酵解促进其耐受应激微环境的去泛素化机制研究

Glioma cells can adapt to the complex and diverse microenvironment, which is one of the fundamental reasons for its refractory. Particularly, it is an important way of glioma cells to withstand stress microenvironment by metabolic reprogramming. Meanwhile, abnormal glycolytic activity is the most common way of metabolism reprogramming in tumor cells. Recent studies have shown that the deubiquification is a key link in regulating glycolysis in tumor cells. In the previous study, the applicant found that YOD1 was significantly elevated in glioma tissues, and elevated glycolysis of glioma cells and promoted the expression of the key glycolysis enzyme HK2 by reducing its ubiquitination level. Therefore, the applicant states the hypothesis that YOD1 regulates the glycolysis of glioma cells by deubiquitination and promotes its tolerance to stress microenvironment. To confirm this hypothesis, this project will demonstrate the role of YOD1 on the biological behavior and glycolysis in glioma cells under stress microenvironment by using in vitro and in vivo studies, and will clarified the mechanism of how YOD1 regulates the deubiquitination of HK2 through immunoprecipitation and ubiquitin quantitative proteomics. Because YOD1 promotes glioma cells to tolerate stress microenvironment by regulating glycolysis, this project will be an important supplement to the theory of new regulatory mechanisms for tumor metabolic reprogramming under stress microenvironment and will provide new ideas and targets for the treatment of gliomas.

胶质瘤细胞能够适应复杂多变微环境是其难治的根本原因之一，其中代谢重编程是胶质瘤细胞耐受应激微环境的重要途径，而糖酵解异常活跃是肿瘤细胞代谢重编程最常见的方式，最新研究表明去泛素化机制是调控糖酵解的重要环节。申请者前期研究发现，在胶质瘤组织中显著升高的去泛素化酶YOD1能够调控糖酵解关键酶HK2的泛素化水平及胶质瘤细胞的糖酵解水平。因此提出YOD1通过去泛素化调控胶质瘤细胞糖酵解促使其耐受应激微环境的假说。为证实假说，本项目将通过体内体外实验证实在应激微环境中去泛素化酶YOD1对胶质瘤细胞生物学行为和糖酵解调控的作用；拟利用泛素化定量蛋白组学，免疫共沉淀等技术阐明YOD1调控HK2等糖酵解关键酶的去泛素化机制，明确去泛素化酶YOD1依赖调控糖酵解促进胶质瘤细胞耐受应激微环境的意义。本项目是对应激微环境下肿瘤代谢重编程的调控机制理论的重要补充，将为胶质瘤的治疗提供新思路和新靶点。

胶质瘤细胞能够适应复杂多变微环境是其难治的根本原因之一，其中代谢重编程是胶质瘤细胞耐受应激微环境的重要途径，而糖酵解异常活跃是肿瘤细胞代谢重编程最常见的方式。本课题组研究发现，在胶质瘤组织中显著升高的去泛素化酶YOD1能够调控糖酵解关键酶HK2的泛素化水平及胶质瘤细胞的糖酵解水平；通过YOD1敲除稳转细胞系的建立，发现以HK2为代表的部分糖酵解代谢酶有表达趋势改变。本项目是对应激微环境下肿瘤代谢重编程的调控机制理论的重要补充，将为胶质瘤的治疗提供新思路和新靶点。.同时本课题组在研究中根据发现的新线索，证实蛋氨酸代谢对于肿瘤代谢重编程具有重要的作用。通过实验证明了在体内外，蛋氨酸剥夺通过下调IL1RN来抑制胶质瘤细胞的增殖，蛋氨酸剥夺或敲除IL1RN诱导的胶质瘤细胞周期阻滞。证实IL1RN是肿瘤相关基因，其表达与胶质瘤患者生存时间呈负相关。证明靶向蛋氨酸剥夺/IL1RN相关基因治疗可能为胶质瘤治疗提供新的方向。.此外课题组在对于肿瘤微环境的研究过程中发现，肿瘤免疫微环境与胶质瘤的恶性进展及治疗耐药密切相关。长链非编码RNA (Long non-coding RNA, lncRNA)在这一过程中起着调节作用。我们通过实验筛选出5个免疫基因相关lncRNAs (AP001007.1、LBX-AS1、MIR155HG、MAPT-AS1、LINC00515)构建风险模型。发现风险评分与患者预后及临床特征相关，与PDL1、TIM-3、B7-H3表达呈正相关。这些lncRNA可能通过细胞因子-细胞因子受体相互作用、补体作用、凝血级联作用等调节肿瘤免疫微环境，在高免疫组可能促进CD8+ T细胞、调节性T细胞、M1巨噬细胞、浸润中性粒细胞活性。我们构建了免疫基因相关lncRNA风险模型，相关风险评分可能成为肿瘤免疫亚型新的生物标志物，为胶质瘤免疫治疗提供分子靶点。..