2. Academic Validation
  3. High-fidelity Cas9-mediated targeting of KRAS driver mutations restrains lung cancer in preclinical models

High-fidelity Cas9-mediated targeting of KRAS driver mutations restrains lung cancer in preclinical models

  • Nat Commun. 2025 Sep 1;16(1):7080. doi: 10.1038/s41467-025-62350-4.
Juan Carlos Álvarez-Pérez # 1 2 3,  Juan Sanjuán-Hidalgo # 1 2,  Alberto M Arenas # 1 2 3,  Ivan Hernández-Navas 4 5,  Maria S Benitez-Cantos 1 3 6,  Alvaro Andrades 1 2 3,  Silvia Calabuig-Fariñas 7 8,  Eloisa Jantus-Lewintre 8 9,  Luis Paz-Ares 5,  Irene Ferrer 4 5,  Pedro P Medina 10 11 12
Affiliations

Affiliations

  • 1 Gene Expression Regulation and Cancer Group (CTS-993). GENYO. Centre for Genomics and Oncological Research: Pfizer-University of Granada-Andalusian Regional Government, Granada, Spain.
  • 2 Department of Biochemistry and Molecular Biology I, Faculty of Sciences, University of Granada, Granada, Spain.
  • 3 Instituto de Investigación Biosanitaria ibs, Granada, Spain.
  • 4 Targeted Therapies for Precision Oncology, Instituto de Investigación Hospital 12 de Octubre (i+12), CIBERONC, Madrid, Spain.
  • 5 H12O-CNIO Lung Cancer Clinical Research Unit, Instituto de Investigación Hospital 12 de Octubre (i+12) & Centro Nacional de Investigaciones Oncológicas (CNIO), CIBERONC, UCM, Madrid, Spain.
  • 6 Department of Biochemistry and Molecular Biology III, Faculty of Medicine, University of Granada, Granada, Spain.
  • 7 Department of Pathology, Universitat de Valencia, Valencia, Spain.
  • 8 Molecular Oncology Lab, Fundación para la Investigación del Hospital General Universitario de Valencia; CIBERONC, Valencia, Spain.
  • 9 Department of Biotechnology, Joint Unit UPV-CIPF Nanomedicine and Disease Mechanisms. Universitat Politècnica de València and Centro de Investigación Príncipe Felipe, Valencia, Spain.
  • 10 Gene Expression Regulation and Cancer Group (CTS-993). GENYO. Centre for Genomics and Oncological Research: Pfizer-University of Granada-Andalusian Regional Government, Granada, Spain. pedromedina@ugr.es.
  • 11 Department of Biochemistry and Molecular Biology I, Faculty of Sciences, University of Granada, Granada, Spain. pedromedina@ugr.es.
  • 12 Instituto de Investigación Biosanitaria ibs, Granada, Spain. pedromedina@ugr.es.
  • # Contributed equally.
PMID: 40890128 DOI: 10.1038/s41467-025-62350-4
Abstract

Missense mutations in the 12th codon of KRAS are key drivers of lung Cancer, with glycine-to-cysteine (G12C) and glycine-to-aspartic acid (G12D) substitutions being among the most prevalent. These mutations are strongly associated with poor survival outcomes. Given the critical role of KRAS in lung Cancer and Other cancers, it remains as a major target for the development of new and complementary treatments. We have developed a CRISPR-High Fidelity (HiFi)-Cas9-based therapy strategy that can effectively and specifically target KRASG12C and KRASG12D mutants, avoiding KRASWT off-targeting and affecting KRAS downstream pathways, thereby significantly reducing tumorgenicity. The delivery of HiFiCas9 components via ribonucleoprotein particles (RNPs) and adenovirus (AdV) effectively abrogates cell viability in KRAS-mutant Non-Small Cell Lung Cancer (NSCLC) preclinical models, including 2D and 3D cell cultures, cell-derived xenografts (CDX), and patient-derived xenograft organoids (PDXO). Our in vitro studies demonstrate that HiFiCas9-based therapy achieves superior KRAS inhibition compared to Sotorasib and effectively circumvents certain resistance mechanisms associated with Sotorasib treatment. Moreover, in vivo delivery using adenoviral particles significantly suppresses tumor growth in preclinical NSCLC models. Collectively, our findings establish HiFiCas9 as an effective therapeutic strategy with promising clinical applications, especially if in vivo delivery methods are further optimized.

Figures
Products
  • Cat. No.
    Product Name
    Description
    Target
    Research Area
  • HY-114277
    99.75%, KRAS G12C Inhibitor
    Ras