Genomic GLO1 deletion from human prostate cancer cells suppresses EMT gene expression and bone metastasis in a bioluminescent SCID mouse model

Glyoxalase 1 (encoded by GLO1 ) is a glutathione-dependent enzyme detoxifying the glycolytic byproduct methylglyoxal (MG), an oncometabolite involved in metabolic reprogramming and tumorigenesis. Prior research has identified GLO1 overexpression as a critical molecular hallmark of prostate cancer progression, and high GLO1 protein levels strongly correlate with advanced histological tumor grade, pathological stage, and early biochemical recurrence. Here, after confirming GLO1 overexpression by TCGA pan-cancer RNA-seq transcriptomic analysis of human tumor samples, we have performed CRISPR/Cas9-mediated genetic deletion of GLO1 from bioluminescent DU145-Luc2 metastatic prostate cancer cells to investigate transcriptomic and phenotypic impact in vitro and in vivo . NanoString nCounter™ (‘PanCancer Progression’ panel) comparative gene expression profiling ( GLO1 _KO versus GLO1 _WT) revealed a significant downregulation of genes and pathways related to epithelial-to-mesenchymal transition (EMT) and cancer progression as a consequence of GLO1 deletion; concordantly, phenotypical screening indicated a pronounced attenuation of matrigel invasiveness observable in GLO1 _KO cells. Likewise, inhibition of GLO1 using a pharmacological inhibitor attenuated the invasive capacity of DU145- GLO1 _WT cells, an effect recapitulated across additional prostate carcinoma cell lines (PC-3, LNCaP). In a bioluminescent SCID mouse bone metastasis model, intracardial injection of DU145-Luc2 isogenic lines [ GLO1 _KO versus GLO1 WT] led to extensive mandibular bone metastasis of GLO1 _WT cells, contrasted by the complete absence of metastasis in response to GLO1 genetic deletion. In summary, our preclinical data are consistent with a molecular role of GLO1 in prostate cancer cell invasiveness and metastasis, warranting further experimental evaluation.