Defining the initial events in oncogenesis and the cellular responses they entrain, even in advance of morphological abnormality, is a fundamental challenge in understanding cancer initiation. As a paradigm to address this, we longitudinally studied the changes induced by loss of the tumor suppressor gene von Hippel Lindau (VHL), which ultimately drives clear cell renal cell carcinoma. Vhl inactivation was directly coupled to expression of a tdTomato reporter within a single allele, allowing accurate visualization of affected cells in their native context and retrieval from the kidney for single-cell RNA-sequencing. This strategy uncovered cell-type specific responses to Vhl inactivation, defined a proximal tubular cell class with oncogenic potential, and revealed longer term adaptive changes in the renal epithelium and the interstitium. Oncogenic cell tagging also revealed markedly heterogeneous cellular effects including time-limited proliferation and elimination of specific cell types. Overall, this study reports an experimental strategy for understanding oncogenic processes in which cells bearing genetic alterations can be generated in their native context, marked, and analyzed over time. The observed effects of loss of Vhl in kidney cells provide insights into VHL tumor suppressor action and development of renal cell carcinoma.

中文翻译：

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致癌细胞标记和单细胞转录组学揭示了肾脏中 Vhl 失活的细胞类型特异性和时间分辨反应

定义肿瘤发生的初始事件及其引发的细胞反应，即使是在形态异常之前，也是理解癌症发生的一个基本挑战。作为解决这个问题的范例，我们纵向研究了肿瘤抑制基因 von Hippel Lindau (VHL) 缺失引起的变化，最终导致透明细胞肾细胞癌。 Vhl 失活直接与单个等位基因内 tdTomato 报告基因的表达耦合，从而可以在其天然环境中准确地观察受影响的细胞，并从肾脏中检索以进行单细胞 RNA 测序。该策略揭示了细胞类型对 Vhl 失活的特异性反应，定义了具有致癌潜力的近端肾小管细胞类别，并揭示了肾上皮和间质的长期适应性变化。致癌细胞标记还揭示了明显的异质性细胞效应，包括限时增殖和特定细胞类型的消除。总体而言，这项研究报告了一种用于理解致癌过程的实验策略，其中携带遗传改变的细胞可以在其天然环境中生成，随着时间的推移进行标记和分析。观察到的肾细胞中 Vhl 缺失的影响为了解 VHL 肿瘤抑制作用和肾细胞癌的发展提供了见解。