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Updated: Jun 30, 2025
Comparing Metastatic Clear Cell Renal Cell Carcinoma Model Established in Mouse Kidney and on Chicken Chorioallantoic Membrane
Published on: February 8, 2020
Oncogenic Cell Tagging and Single-Cell Transcriptomics Reveal Cell Type-Specific and Time-Resolved Responses to Vhl
Samvid Kurlekar1,2, Joanna D C C Lima1,2, Ran Li1
1Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom.
Abstract:
Defining the initial events in oncogenesis and the cellular responses they entrain, even in advance of morphologic 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.
Significance:
Single-cell analysis of heterogeneous and dynamic responses to Vhl inactivation in the kidney suggests that early events shape the cell type specificity of oncogenesis, providing a focus for mechanistic understanding and therapeutic targeting.
Insights
Loss of the von Hippel Lindau (VHL) gene initiates kidney cancer by altering specific cell types. This study visualizes and analyzes these early oncogenic events using single-cell sequencing.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Understanding cancer initiation requires defining early oncogenic events and cellular responses.
- The von Hippel Lindau (VHL) tumor suppressor gene is crucial in clear cell renal cell carcinoma development.
Purpose of the Study:
- To investigate the cellular and molecular changes following VHL gene loss in the kidney.
- To identify specific cell types involved in VHL-deficient renal cell carcinoma initiation.
- To develop a method for tracking and analyzing cells with genetic alterations in their native context.
Main Methods:
- Longitudinal study of VHL inactivation in kidney cells.
- Utilized a tdTomato reporter coupled to VHL allele for cell marking.
- Employed single-cell RNA sequencing to analyze cellular responses.
- Investigated adaptive changes in renal epithelium and interstitium.
Main Results:
- VHL inactivation induced cell type-specific responses in the kidney.
- Identified a proximal tubular cell population with oncogenic potential.
- Observed heterogeneous cellular effects, including transient proliferation and cell elimination.
- Revealed adaptive changes in the renal epithelium and interstitium over time.
Conclusions:
- Early cellular responses to VHL inactivation shape oncogenesis specificity.
- The developed experimental strategy enables in-situ analysis of oncogenic processes.
- Findings provide insights into VHL tumor suppressor function and renal cell carcinoma development.
- Highlights potential therapeutic targets based on cell type-specific responses.
