Background: Modeling precancerous stages holds the promise to understand early transformation events, thereby offering the potential for personalized, targeted treatment. Because cancer hijacks developmental pathways, precancerous stages could potentially be modeled by reprogramming cancer cells to an induced pluripotent stem cell state and subsequently differentiating them to the target organs using organoid models. Methods: We attempted reprogramming of patient-derived clear cell renal cell carcinoma (ccRCC) cell lines and adjacent normal renal epithelial cell lines using lentivirus or episomal reprogramming vectors. Results: The cancer cells failed to reprogram while the adjacent normal cells reprogrammed with high efficiency. The von Hippel–Lindau factor (VHL) gene was re-expressed in ccRCC cells in an attempt to restore the wild-type phenotype and restore reprogramming. The VHL gene is the major tumor suppressor in ccRCC pathogenesis and a conductor of oxidative-glycolytic glucose metabolism. While its re-expression did restore the epithelial phenotype and oxidative regulation of ccRCC cells, they still failed to stably reprogram. With an optimized reprogramming workflow, VHL-corrected ccRCC cells generate NANOG+ cells; however, they remained dependent on the ectopic expression of the reprogramming factors. Conclusions: We concluded that while VHL expression is necessary for cellular reprogramming of ccRCC cells, other genetic lesions in the ccRCC cells could be preventing the stabilization of the pluripotent state.
背景：对癌前阶段进行建模有助于理解早期转化事件，从而为个性化靶向治疗提供可能。由于癌症劫持了发育通路，通过将癌细胞重编程为诱导多能干细胞状态，并随后利用类器官模型将其分化为目标器官，有望实现对癌前阶段的建模。方法：我们尝试使用慢病毒或附加体重编程载体对患者来源的透明细胞肾细胞癌（ccRCC）细胞系及邻近正常肾上皮细胞系进行重编程。结果：癌细胞未能成功重编程，而邻近正常细胞则实现了高效重编程。为恢复野生型表型并实现重编程，我们在ccRCC细胞中重新表达了VHL基因。VHL基因是ccRCC发病机制中的主要抑癌基因，也是氧化-糖酵解葡萄糖代谢的调控因子。尽管其重新表达确实恢复了ccRCC细胞的上皮表型和氧化代谢调控，但这些细胞仍未能实现稳定重编程。通过优化重编程流程，VHL校正的ccRCC细胞能够产生NANOG阳性细胞，但这些细胞仍依赖于外源性重编程因子的表达。结论：我们得出结论，虽然VHL表达是ccRCC细胞重编程的必要条件，但ccRCC细胞中的其他遗传损伤可能阻碍了多能状态的稳定。
VHL Gene Restoration Supports RCC Reprogramming to iPSCs but Does Not Ensure Line Stability
肿瘤(癌症)患者之家