Abstract 3171: Multitargeted kinase inhibitor LCI139 overcomes chemotherapy resistance in patient-derived non small cell lung cancer organoids by harnessing intrinsic and extrinsic apoptosis

Abstract Introduction: In-silico designed multitarget inhibitor LCI139 (PI3K-CDK4/6-CDK9-AURKA/B) was engineered to overcome resistance inherent to single-target agents by simultaneously targeting tumor metabolism and inducing robust apoptosis. This compound demonstrated nanomolar potency against NSCLC in vitro. We evaluated efficacy in NSCLC cell lines and patient-derived tumor organoids (PTOs)—biofabricated spherical constructs recapitulating cellular matrix of native tumor tissue. Methods: NSCLC cell lines (NCI-H1703, NCI-H1781) and PTOs generated from unsorted lung tumor suspensions in PEPGEL matrix (n=7) were utilized under IRB-approved protocol. Treatments included LCI139 (0.25-1μM), chemotherapeutics (carboplatin, gemcitabine, pemetrexed, cisplatin), and single-agent inhibitors: PI3K (Buparlisib), CDK4/6 (Ribociclib), CDK9 (AZD4573), AURKA (Barasertib), AURKB (Alisertib). Cell lines were co-treated with inhibitors of (carbenoxolone), Caspase (Casp)-8 (Z-IETD-FMK), Casp-9 (Z-LEHD-FMK), or AMPK activator (PF-06409577). Viability (propidium iodide, CellTiter-Glo), cell cycle (DNA dye) and apoptosis (Annexin-V/7-AAD, Casp-3/7/8/9 activity) were measured. Target inhibition was evaluated immunoblotting for pAkt, pRb, pRNA Pol II, MCL-1, pAMPK, pFOXO3, and p53. Results: LCI139 achieved nanomolar IC50 against H1703 cells (48hr) and demonstrated dose-dependent PTO cytotoxicity: 49% reduction at 0.25μM, 62% at 0.5μM, 78.4% at 1μM (all p0.0001). By comparison, 1 μM single-agent inhibitors reduced PTO viability by: AURKA (34%), AURKB (24%), PI3K (29%), CDK9 (50.9%) all p0.0001; CDK4/6 (6.9%, p=0.43). LCI139 overcame significant carboplatin resistance (IC50: 10.25-11.69μM) and outperformed standard chemotherapeutics: gemcitabine (29.2%), cisplatin (28.2%), pemetrexed (no activity). LCI139 activated caspases 8, 9, 3/7 in cells and PTOs. Selective Casp-8 or Casp-9 inhibition diminished but didn't abrogate cytotoxicity. Western blotting confirmed dose-dependent target inhibition: decreased pAKT, pRb, MCL-1, pRNAPol II, and p53 modulation. LCI139 induced metabolic stress via pAMPK-pFoxO3 activation. FoxO3 inhibition diminished LCI139 potency against sensitive H1703 cells. AMPK activation enhanced potency in p53-proficient H1703 and sensitized p53-deficient H1781 cells. Conclusions: LCI139 demonstrates superior anti-tumor activity versus standard chemotherapeutics in resistant NSCLC through dual activation of intrinsic (cell cycle arrest-induced) and extrinsic (metabolic stress-induced) apoptotic pathways. Efficacy in both cell lines and PTOs validates this micro-physiological platform for drug development, supporting translational potential of multitargeted kinase inhibition for overcoming therapeutic resistance, as aligned with FDA guidance. Citation Format: Anna Ivanina Foureau, Nadeem Wajih, Cody C. McHale, Hailey L. Dryden, Sara Muhiczukic, Dhananjaya Pal, Bharath Yada, David Foureau, Fei Guo, Raj Dhupar, Donald Durden, Konstantinos Votanopoulos, Shay Soker, Eleftherios Markis, Kathryn Mileham. Multitargeted kinase inhibitor LCI139 overcomes chemotherapy resistance in patient-derived non small cell lung cancer organoids by harnessing intrinsic and extrinsic apoptosis abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 3171.