Introduction: Lung cancer is the leading cause of cancer-related mortality worldwide, with non-small cell lung cancer (NSCLC) accounting for nearly 85% of cases. Mutations in the STK11 gene are present in approximately 30% of NSCLC and are strongly associated with tumor progression and resistance to therapy. To better understand the mechanisms underlying STK11 deficiency, advanced in vitro models are required. Organoids derived from lung epithelial cells offer a physiologically relevant three-dimensional system that recapitulates the architecture of lung tissue.
Methods: Human lung epithelial organoids were cultured from stem/progenitor cells. A lentiviral vector encoding CRISPR-Cas9 and a single guide RNA targeting STK11 was used for stable transduction. Organoids were analyzed for gene editing efficiency using PCR and sequencing. Functional assays included cell proliferation, metabolic profiling, and morphological characterization to evaluate tumor-like phenotypes induced by STK11 knockout.
Results: CRISPR-mediated knockout of STK11 in lung organoids resulted in significant metabolic reprogramming, enhanced proliferation, and disruption of normal epithelial polarity. These phenotypic changes resembled early tumorigenic events observed in NSCLC. Importantly, STK11-deficient organoids exhibited resistance to energy stress, consistent with clinical data from lung cancer patients.
Conclusion: Combining organoid technology with CRISPR-Cas9 genome editing enables precise modeling of lung cancer driver mutations. Targeting STK11 in lung organoids provides a robust experimental platform for dissecting mechanisms of tumor initiation, progression, and therapy resistance. This approach can accelerate the discovery of novel therapeutic strategies against NSCLC.