Abstract 6658: Inhibiting glycolysis for the treatment of HGF/MET/TWIST1 driven NSCLC brain metastases

Abstract Non-small cell lung cancer (NSCLC) has the highest incidence of brain metastases (BM), with ∼40% of patients developing BM. We identified a significant enrichment of MET amplification (amp) in NSCLC BM compared to primary lung and liver metastases. MET, a receptor tyrosine kinase, and its ligand, hepatocyte growth factor (HGF), promote proliferation, epithelial-mesenchymal transition, angiogenesis, metastasis and metabolic reprogramming through glycolysis. We have found that MET amp NSCLC BM and MET altered cells have increased glycolysis compared to MET wildtype (WT) BM and cells. We have also shown that TWIST1 is regulated by the HGF/MET pathway, essential for MET-driven NSCLC and MET TKI resistance and responsible for HGF/MET-driven hexokinase II (HK2) expression in MET altered cells. HK2, a critical node for metabolic regulation, catalyzes the first step in glucose metabolism. Thus, we hypothesize that targeting glycolysis in MET-driven NSCLC may be an effective therapeutic strategy. We have previously shown that metastatic H1993 (MET amp) cells which were derived from the same patient as primary lung H2073 (MET WT) cells exhibit increased glycolysis and increased glycolytic dependency. We have now found that TWIST1 was sufficient to drive glycolytic dependency as TWIST1 overexpression (OE) in H2073 cells enhanced sensitivity to the HK2 inhibitor 2-deoxyglucose (2DG), while TWIST1 knockdown in H2073 cells decreased sensitivity to 2DG. Interestingly, TWIST1 OE in H1993 cells increased HK2 levels, leading to 2DG resistance but increased sensitivity to the downstream glycolytic inhibitor, PFK158 with similar or decreased sensitivity to oxidative phosphorylation (OXPHOS) inhibitors. To examine whether glycolytic inhibitors would be effective at the time of MET TKI resistance, we used our MET altered models of acquired MET TKI resistance, H1993 CR and UW-21 CR. H1993 CR (low HK2) cells exhibited reduced sensitivity to 2DG but increased sensitivity to PFK158 and AT-101, while UW-21 CR (high HK2) cells showed reduced sensitivity to all three inhibitors compared to parental cells. We then examined whether glycolytic inhibition would be effective against MET amp driven EGFR TKI resistance. HCC827R2 (acquired MET amp) cells demonstrated increased HK2 protein expression and heightened sensitivity to 2DG and AT-101 compared to HCC827 cells. In a more relevant ex vivo brain slice model, we found that H1993 brain colonization was inhibited by 2DG and AT-101 but not OXPHOS inhibitors, while H2073 colonization showed limited response to glycolytic inhibitors but more sensitivity to OXPHOS inhibitors, consistent with our previous findings in patient BM. Remarkably, H1993 CR cells which were resistant to glycolytic inhibition in vitro were sensitive to 2DG ex vivo. These findings suggest that MET-driven glycolytic dependency plays a significant role in BM, highlighting glycolysis as a potential therapeutic target. Citation Format: Xiaxuan Wu, Jason Chen, Thomas Hintelmann, Andrew Zhao, Joseph Dosch, Rahul Desai, Purva H. Rumde, Kasey R. Cargill, Riyue Bao, Vinod Kumar, Laura P. Stabile, Timothy F. Burns. Inhibiting glycolysis for the treatment of HGF/MET/TWIST1 driven NSCLC brain metastases [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 6658.