# Manipulation with Mutational Status of VHL Regulates Hypoxic Metabolism and Pro-Angiogenic Phenotypes in ccRCC Caki-1 Cells

## TL;DR

This study shows that restoring the VHL gene in kidney cancer cells reduces aggressive traits and hypoxia-like metabolism, offering a potential new treatment approach for ccRCC.

## Contribution

The study demonstrates that VHL restoration can reverse aggressive tumor phenotypes and hypoxic metabolism in ccRCC cells.

## Key findings

- VHL inactivation increases hypoxia-like pathways and anaerobic glycolysis in Caki-1 cells.
- Restoring VHL reverses these metabolic changes and modulates angiogenesis-related gene expression.
- VHL-positive and -negative cells show distinct proportions of benign and aggressive subpopulations.

## Abstract

Clear cell renal cell carcinoma (ccRCC), accounting for 80–90% of renal malignancies, is frequently driven by VHL inactivation—either through mutation or promoter hypermethylation—resulting in constitutive HIF2α activation and pseudohypoxic signaling. VHL gene inactivation is a hallmark of von Hippel–Lindau syndrome, a hereditary disorder predisposing patients to ccRCC and other tumors, underscoring its central role in disease pathogenesis. While VHL dysfunction promotes aggressive tumor phenotypes, the therapeutic potential of VHL restoration remains underexplored. Here, using the Cas9 induced VHL-mutation in the Caki-1 cell line model, we demonstrate that VHL inactivation augments hypoxia-like pathways and enhances anaerobic glycolysis. Rescue of functional VHL reversed these activation patterns and modulated the expression of genes associated with angiogenesis. Using single cell transcriptomics, we show that the VHL-positive and -negative Caki-1 cells are characterized with different proportions of benign and aggressive cells as seen by analysis of specific gene expression. Furthermore, the identified angiogenesis-related genes were linked to affect clinical outcomes in ccRCC patients, suggesting that VHL restoration may mitigate high-risk molecular features.