Multiplexed screens identify RAS paralogues HRAS and NRAS as suppressors of KRAS-driven lung cancer growth

  • Nat Cell Biol. 2023 Jan 12. doi: 10.1038/s41556-022-01049-w.
Rui Tang  #  1 ,  Emily G Shuldiner  #  2 ,  Marcus Kelly  3  4 ,  Christopher W Murray  3 ,  Jess D Hebert  1 ,  Laura Andrejka  1 ,  Min K Tsai  1  3 ,  Nicholas W Hughes  1 ,  Mitchell I Parker  5  6 ,  Hongchen Cai  1 ,  Yao-Cheng Li  7 ,  Geoffrey M Wahl  7 ,  Roland L Dunbrack  5 ,  Peter K Jackson  3  4 ,  Dmitri A Petrov  2  3  8 ,  Monte M Winslow  9  10  11
Affiliations
  • 1. Department of Genetics, Stanford University School of Medicine, Stanford, CA, USA.
  • 2. Department of Biology, Stanford University, Stanford, CA, USA.
  • 3. Cancer Biology Program, Stanford University School of Medicine, Stanford, CA, USA.
  • 4. Baxter Laboratories, Stanford University School of Medicine, Stanford, CA, USA.
  • 5. Molecular Therapeutics Program, Institute for Cancer Research, Fox Chase Cancer Center, Philadelphia, PA, USA.
  • 6. Molecular and Cell Biology and Genetics Program, Drexel University College of Medicine, Philadelphia, PA, USA.
  • 7. Gene Expression Laboratory, The Salk Institute for Biological Studies, La Jolla, CA, USA.
  • 8. The Chan Zuckerberg BioHub, San Francisco, CA, USA.
  • 9. Department of Genetics, Stanford University School of Medicine, Stanford, CA, USA. [email protected].
  • 10. Cancer Biology Program, Stanford University School of Medicine, Stanford, CA, USA. [email protected].
  • 11. Department of Pathology, Stanford University School of Medicine, Stanford, CA, USA. [email protected].
  • # Contributed equally.
PMID: 36635501 DOI: 10.1038/s41556-022-01049-w
Abstract

Oncogenic KRAS mutations occur in approximately 30% of lung adenocarcinoma. Despite several decades of effort, oncogenic KRAS-driven lung Cancer remains difficult to treat, and our understanding of the regulators of Ras signalling is incomplete. Here to uncover the impact of diverse KRAS-interacting proteins on lung Cancer growth, we combined multiplexed somatic CRISPR/Cas9-based genome editing in genetically engineered mouse models with tumour barcoding and high-throughput barcode Sequencing. Through a series of CRISPR/Cas9 screens in autochthonous lung Cancer models, we show that HRAS and NRAS are suppressors of KRASG12D-driven tumour growth in vivo and confirm these effects in oncogenic KRAS-driven human lung Cancer cell lines. Mechanistically, Ras paralogues interact with oncogenic KRAS, suppress KRAS-KRAS interactions, and reduce downstream ERK signalling. Furthermore, HRAS and NRAS mutations identified in oncogenic KRAS-driven human tumours partially abolished this effect. By comparing the tumour-suppressive effects of HRAS and NRAS in oncogenic KRAS- and oncogenic BRAF-driven lung Cancer models, we confirm that Ras paralogues are specific suppressors of KRAS-driven lung Cancer in vivo. Our study outlines a technological avenue to uncover positive and negative regulators of oncogenic KRAS-driven Cancer in a multiplexed manner in vivo and highlights the role Ras paralogue imbalance in oncogenic KRAS-driven lung Cancer.

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