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Updated: Sep 9, 2025
Detection of Lung Tumor Progression in Mice by Ultrasound Imaging
Published on: February 27, 2020
High-fidelity Cas9-mediated targeting of KRAS driver mutations restrains lung cancer in preclinical models
Juan Carlos Álvarez-Pérez1,2,3, Juan Sanjuán-Hidalgo1,2, Alberto M Arenas1,2,3
1Gene Expression Regulation and Cancer Group (CTS-993). GENYO. Centre for Genomics and Oncological Research: Pfizer-University of Granada-Andalusian Regional Government, Granada, Spain.
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.
Insights
CRISPR-HiFi-Cas9 therapy effectively targets KRAS G12C and G12D mutations in lung cancer. This novel approach reduces tumor growth and shows promise against Sotorasib resistance.
Area of Science:
- Oncology
- Molecular Biology
- Gene Editing
Background:
- KRAS mutations, particularly G12C and G12D, are prevalent drivers in lung cancer, associated with poor prognosis.
- KRAS is a critical therapeutic target in various cancers, necessitating innovative treatment strategies.
Purpose of the Study:
- To develop and evaluate a CRISPR-High Fidelity (HiFi)-Cas9-based gene editing strategy targeting KRAS G12C and G12D mutations.
- To assess the efficacy and specificity of this therapy in preclinical lung cancer models and compare it with existing treatments.
Main Methods:
- Utilized CRISPR-HiFi-Cas9 technology delivered via ribonucleoprotein particles (RNPs) and adenovirus (AdV).
- Tested the therapy in various Non-Small Cell Lung Cancer (NSCLC) preclinical models, including 2D/3D cultures, CDX, and PDXO.
- Conducted in vitro comparisons with Sotorasib and evaluated in vivo tumor suppression.
Main Results:
- HiFi-Cas9 therapy specifically targeted KRAS G12C/G12D mutants, avoiding wild-type KRAS, and reduced tumorgenicity.
- Demonstrated effective abrogation of cell viability in KRAS-mutant NSCLC models.
- Achieved superior KRAS inhibition compared to Sotorasib in vitro and circumvented resistance mechanisms. Adenoviral delivery significantly suppressed tumor growth in vivo.
Conclusions:
- CRISPR-HiFi-Cas9 represents a potent and specific therapeutic strategy for KRAS-mutant lung cancers.
- The therapy shows promise in overcoming resistance to current treatments like Sotorasib.
- Further optimization of in vivo delivery methods could enhance clinical applicability.
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