Targeting the PYGL with chicoric acid to normalize glucose metabolism to inhibit progress in NSCLC.

Abstract

e15096 Background: Glucose metabolism reprogramming often drives the malignant progression of tumors, and targeted therapies against aberrant tumor metabolism have emerged as promising strategies to combat tumor growth. Liver glycogen phosphorylase (PYGL) is a key protein in the regulation of glucose metabolic homeostasis for the breakdown of glycogen to glucose. Chicoric acid (CA) is a pharmacologically active natural compound extracted from chicory and Echinacea purpurea with anti-inflammatory, antioxidant and apoptosis-inducing properties. However, the role of CA in cancer therapy remains unclear. Methods: Bioinformatics screening and analyses were performed to identify hub gene. The relationship between PYGL expression and clinicopathological features in non-small cell lung cancer (NSCLC) was assessed by immunohistochemistry. The effects of PYGL on cell proliferation and apoptosis were evaluated in vitro and in vivo by CCK-8, flow cytometry and xenograft model. PAS glycogen staining, 2-NDBG molecular probe and seahorse were utilized to measure the regulation of PYGL to glucose and glycogen metabolism. Virtual screening, molecular docking, cellular thermal shift assay (CETAS) and surface plasmon resonance (SPR) were performed to find and verify the target drugs of PYGL. Moreover, bind sites of CA and PYGL were demonstrated by point mutation, the Native-PAGE was conducted to detect protein activity. The transcriptomic and other biological methods were used to explore related pathways. Results: In this study, we founded that PYGL can be used as a potential marker for the diagnosis and prognosis in NSCLC. Our results suggested that PYGL dependents on enzymatic activity to accelerate NSCLC cell growth by facilitating glycolysis. Mechanistically, we revealed that E2F1 enhances PYGL transcription, and then PYGL upregulated LDHA through the AMPK/mTOR signaling axis, leading to glycolytic reprogramming and malignance in NSCLC. Therapeutically, we identified that CA as a small molecule drug of PYGL strongly binds to PYGL ( K d : 10 μmol/L) through specific amino acid residues (R248A, E274A, and S277A), which crippled phosphorylation level PYGL and increased glycogen storage and thereby inhibiting glycolysis and progression of tumor cell. Conclusions: Our results suggest that PYGL plays a carcinogenic role by reshaping glucose metabolism. CA interferes phosphorylation of PYGL to attenuate progression in NSCLC, it may be a promising therapy for patients with high PYGL expression warrants further clinical studies.