Closely related PPP genes were obtained from the TCGA and GEO databases. The effect of ATP13A2 on CRC cell proliferation was evaluated by performing in vitro assays. The connection between the PPP and ATP13A2 was explored by assessing proliferation and antioxidative stress. The molecular mechanism by which ATP13A2 regulates the PPP was investigated using chromatin immunoprecipitation and dual luciferase experiments. The clinical therapeutic potential of ATP13A2 was explored using patient-derived xenograft (PDX), patient-derived organoid (PDO) and AOM/DSS models.
We identified ATP13A2 as a novel PPP-related gene. ATP13A2 deficiency inhibited CRC growth and PPP activity, as manifested by a decrease in the levels of PPP products and an increase in reactive oxygen species levels, whereas ATP13A2 overexpression induced the opposite effect. Mechanistically, ATP13A2 regulated the PPP mainly by affecting phosphogluconate dehydrogenase (PGD) mRNA expression. Subsequent studies showed that ATP13A2 overexpression promoted TFEB nuclear localization by inhibiting the phosphorylation of TFEB, thereby enhancing the transcription of PGD and ultimately affecting the activity of the PPP. Finally, ATP13A2 knockdown inhibited CRC growth in PDO and PDX models. ATP13A2- /- mice had a lower CRC growth capacity than ATP13A2+/+ in the AOM/DSS model.Our findings revealed that ATP13A2 overexpression-driven dephosphorylation of TFEB promotes PPP activation by increasing PGD transcription, suggesting that ATP13A2 may serve as a potential target for CRC therapy.
方法:从TCGA和GEO数据库获得密切相关的PPP基因。通过进行体外测定来评估ATP13A2对CRC细胞增殖的影响。通过评估增殖和抗氧化应激来探索PPP和ATP13A2之间的联系。使用染色质免疫沉淀和双荧光素酶实验研究了ATP13A2调节PPP的分子机制。使用患者来源的异种移植物(PDX)探索了ATP13A2的临床治疗潜力,患者来源的类器官(PDO)和AOM/DSS模型。
结果:我们鉴定了ATP13A2是一种新的PPP相关基因。ATP13A2缺乏抑制CRC生长和PPP活性,表现为PPP产品水平的降低和活性氧水平的增加,而ATP13A2过表达诱导了相反的作用。机械上,ATP13A2主要通过影响磷酸葡萄糖酸脱氢酶(PGD)mRNA表达来调控PPP。随后的研究表明,ATP13A2过表达通过抑制TFEB的磷酸化促进TFEB的核定位,从而增强PGD的转录并最终影响PPP的活性。最后,ATP13A2敲低抑制PDO和PDX模型中的CRC生长。ATP13A2-/-小鼠在AOM/DSS模型中具有比ATP13A2+/+更低的CRC生长能力。我们的发现揭示了ATP13A2过表达驱动的TFEB去磷酸化通过增加PGD转录促进PPP激活,提示ATP13A2可能成为CRC治疗的潜在靶点.