ARRB1 Regulates Metabolic Reprogramming to Promote Glycolysis in Stem Cell-Like Bladder Cancer Cells

SIMPLE SUMMARY: Bladder cancer (BC) ranks second in incidence and mortality among all genitourinary cancers. The high recurrence of BC is attributed to the presence of cancer stem cells (CSCs), which are the driving force behind tumor growth. Increasing evidence suggests that stem cells exhibit a distinct metabolic program compared to differentiated cells. Understanding their metabolic preference for maintaining stem cell properties is essential for developing novel therapeutics targeting CSCs. The current work shows for the first time that the scaffold protein β-arrestin1 (ARRB1) functions as a metabolic switch regulating the metabolic reprogramming of CSC-like cells towards glycolysis by regulating the mitochondrial pyruvate carrier MPC1 and glucose transporter GLUT1. The balance between glycolysis and oxidative phosphorylation plays a crucial role in regulating the fate of stem cells. Our findings will potentially open new therapeutic avenues for targeting bladder cancer cells and/or the CSC-like cells within aggressive bladder tumors. ABSTRACT: β-arrestin 1 (ARRB1) is a scaffold protein that regulates signaling downstream of G protein-coupled receptors (GPCRs). In the current work, we investigated the role of ARRB1 in regulating the metabolic preference of cancer stem cell (CSC)-like cells in bladder cancer (BC). We show that ARRB1 is crucial for spheroid formation and tumorigenic potential. Furthermore, we measured mitochondrial respiration, glucose uptake, glycolytic rate, mitochondrial/glycolytic ATP production and fuel oxidation in previously established ARRB1 knock out (KO) cells and corresponding controls. Our results demonstrate that depletion of ARRB1 decreased glycolytic rate and induced metabolic reprogramming towards oxidative phosphorylation. Mechanistically, the depletion of ARRB1 dramatically increased the mitochondrial pyruvate carrier MPC1 protein levels and reduced the glucose transporter GLUT1 protein levels along with glucose uptake. Overexpression of ARRB1 in ARRB1 KO cells reversed the phenotype and resulted in the upregulation of glycolysis. In conclusion, we show that ARRB1 regulates the metabolic preference of BC CSC-like cells and functions as a molecular switch that promotes reprogramming towards glycolysis by negatively regulating MPC1 and positively regulating GLUT1/ glucose uptake. These observations open new therapeutic avenues for targeting the metabolic preferences of cancer stem cell (CSC)-like BC cells.