TY  - EJOU
AU  - Lin, Kuan-Hao 
AU  - Lin, Yu-Ju 
AU  - Hong, Yu-Bin 
AU  - Hsu, Meng-Huai 
AU  - Liao, Zhen-Xiang 
AU  - Chang, Shuo-Yu 
AU  - Yuh, Chiou-Hwa 

TI  - Glutamic Acid–Chelated Cobalt Stabilizes G-Quadruplexes and Selectively Suppresses Hepatocellular Carcinoma Growth
T2  - Oncology Research

PY  - 2026
VL  - 34
IS  - 4
SN  - 1555-3906

AB  -  <b>Objectives:</b> Hepatocellular carcinoma (HCC) has limited systemic options with substantial toxicity. G-quadruplex (G4) structures in oncogene promoters are attractive but challenging drug targets. This study aimed to determine whether glutamic acid–chelated cobalt (GACC) is a G4-active scaffold with anti-HCC efficacy and favorable <i>in vivo</i> safety, and whether an AI-guided phenotypic response surface (PRS) can optimize less toxic combinations. <b>Methods:</b> Anticancer activity was tested in HCC cell lines (PLC/PRF/5, Hep3B, HepG2) and non-transformed THLE-2 hepatocytes (CCK-8, IC<sub>50</sub>). <i>In vivo</i> safety/efficacy were assessed in zebrafish embryo toxicity assays, a Hep3B xenograft model, and a tert-overexpressing transgenic zebrafish model, with hepatotoxicity monitored in a liver-fluorescent reporter line. Target engagement was examined by docking, native PAGE, a KRAS promoter G4 DNA polymerase stop assay, BG4 immunofluorescence, and KRAS qPCR. PRS was used to optimize GACC–metformin–regorafenib combinations. <b>Results:</b> GACC reduced HCC viability (IC<sub>50</sub> ~86–115 µM) and showed low embryotoxicity (IC<sub>50</sub> 6.87 mM). In zebrafish xenografts, GACC (50 µM) reduced Hep3B tumor fluorescence by ~90% without detectable hepatotoxicity, whereas sorafenib decreased liver size/fluorescence. In tert-overexpressing zebrafish, GACC suppressed proliferation and Wnt/β-catenin–associated transcripts and reduced mitotic figures and nuclear atypia. Mechanistically, GACC increased KRAS promoter polymerase stalling, enhanced nuclear G4 signal, and reduced KRAS transcripts. PRS identified an off-grid triple combination that reduced PLC/PRF/5 viability to 19% while maintaining THLE-2 viability at 52% and preserving zebrafish development. <b>Conclusion:</b> GACC is a G4-active cobalt–glutamate scaffold with anti-HCC activity and favorable zebrafish safety, and a zebrafish-plus-PRS workflow enables rational, less toxic combination design.
KW  - Liver cancer; glutamic acid cobalt chelate; <i>tert</i> transgenic zebrafish; G-quadruplex stabilization; phenotypic response surface; <i>KRAS</i> promoter; zebrafish xenograft

DO  - 10.32604/or.2026.074144