Abstract 6567: Metabolic reprogramming of pentose phosphate pathway mediated by small extracellular vesicles facilitates tumor metastasis and drug resistance in HCC

Abstract Emerging evidence have proved that the interplay between tumor and its microenvironment facilitate tumor metastasis, including the hepatocellular carcinoma (HCC). Therefore, identifying novel target for HCC metastasis and is essential to improve the survival. In recent years, extracellular vesicles (EV) mediated metabolic regulation has been a growing interest in cancer metabolism. For instance, the ectosomal pyruvate kinase M2 isoform (PKM2) was found to facilitate glycolysis reprogramming in monocyte-to-macrophage differentiation and tumor microenvironment remodeling. Compared to directly receiving metabolite or substance from the EV, we proposed that the tumor cells are more likely to obtain proteomics alteration to exert long-term metabolic reprogramming. EVs were extracted from the metastatic and non-metastatic HCC cells. The DEPs were identified by LC-MS/MS. Meanwhile, the metabolomic screening will also be conducted to clarify the differences of metabolite which the HCC cells produced after the treatment of sEVs which derived from metastatic and non-metastatic HCC cell. The expression of G6PD was knockdown by shRNA. The plate cloning and transwell assays were conducted to evaluate the effect of G6PD on the metastasis. Besides, parallel validation of G6PD inhibitors was performed. The expression of gene was validated by real-time PCR. The MTT assay was conducted to test IC50 of sorafenib resistance HCC cells. Western blot was performed to measure expression of different proteins. Our study found that key enzyme involved in pentose phosphate pathway (PPP) was extremely enriched in metastatic HCC derived small extracellular vesicles (sEVs). Inhibition the enzymic activity of sEVs-derived G6PD significantly reduce the invasion and metastasis of HCC tumor cells. Besides, the sEV-G6PD was associated with activation of cancer-associated fibroblast. Moreover, sEV derived PPP enzymes enhanced sorafenib resistance. In conclusion, PPP enzymes derived from sEV in metastatic HCC cells promotes tumor metastasis and sorafenib resistance. Citation Format: Xiaoxin Zhang, Zhixian Chen, Jiaqi Lai, Judy Wai Ping Yam. Metabolic reprogramming of pentose phosphate pathway mediated by small extracellular vesicles facilitates tumor metastasis and drug resistance in HCC [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 6567.