GLDC interacts with VPS34 to inhibit tumorigenesis and epithelial-mesenchymal transition in hepatocellular carcinoma

Abstract

Hepatocellular carcinoma (HCC) accounts for 90% of primary liver cancer with high mortality and limited therapeutic strategy. Glycine decarboxylase (GLDC) is the key limiting enzyme in glycine breakdown metabolism and acts as oncogene or tumor suppressor to impact tumor onset and progression in a context dependent manner. However, the underlying mechanism of GLDC on autophagy and progression is largely unexplored in HCC. Here, we showed that GLDC overexpression inhibited cell proliferation, cell migration and promoted cell senescence and autophagy in HCC. Intriguingly, induced GLDC remarkably attenuated epithelial-mesenchymal transition (EMT) progress and tumor growth in vitro and in vivo. Mechanically, we demonstrated that GLDC upregulated VPS34 protein and enhanced its interaction with VPS34, thus promoting the association of VPS34 with Beclin1/ATG14 complex and autophagy induction in HCC. Importantly, GLDC acetylation at K514 promoted interaction of GLDC-VPS34, whereas GLDC acetylation-dead mutant K514R abolished their binding. Furthermore, GLDC protein was decreased in HCC tissues compared with para-tumor tissues and reduced GLDC was significantly correlated with poor prognosis of patients. In conclusion, we unveil the key regulatory role of GLDC in autophagy and HCC progression through VPS34 and provide a potential strategy for HCC therapy.

Keywords

GLDC / Acetylation / VPS34 / Epithelial-mesenchymal transition / HCC

References

[1] F. Bray, M. Laversanne, H. Sung, J. Ferlay, R.L. Siegel, I. Soerjomataram, A. Jemal, Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries, CA Cancer J. Clin. 74 (3) (2024) 229-263, https://doi.org/10.3322/caac.21834.

[2] C. Yang, H. Zhang, L. Zhang, A.X. Zhu, R. Bernards, W. Qin, C. Wang, Evolving therapeutic landscape of advanced hepatocellular carcinoma, Nat. Rev. Gastro. Hepat. 20 (4) (2023) 203-222, https://doi.org/10.1038/s41575-022-00704-9.

[3] A. Huang, X. Yang, W. Chung, A.R. Dennison, J. Zhou, Targeted therapy for hepatocellular carcinoma, Signal Transduct. Tar. 5 (1) (2020) 146, https://doi.org/10.1038/s41392-020-00264-x.

[4] J.M. Llovet, R. Montal, D. Sia, R.S. Finn, Molecular therapies and precision medicine for hepatocellular carcinoma, Nat. Rev. Clin. Oncol. 15 (10) (2018) 599-616,https://doi.org/10.1038/s41571-018-0073-4.

[5] Y. Wang, B. Deng, Hepatocellular carcinoma: molecular mechanism, targeted therapy, and biomarkers, Cancer Metast. Rev. 42 (3) (2023) 629-652, https://doi.org/10.1007/s10555-023-10084-4.

[6] K.A. Aguayo-Ceron, F. Sanchez-Muñoz, R.A. Gutierrez-Rojas, L.N. Acevedo-Villavicencio, A.V. Flores-Zarate, F. Huang, A. Giacoman-Martinez, S. Villafaña, R. Romero-Nava, Glycine: the smallest anti-inflammatory micronutrient, Int. J. Mol. Sci. 24 (14) (2023) 11236, https://doi.org/10.3390/ijms241411236.

[7] S.L. Geeraerts, E. Heylen, K. De Keersmaecker, K.R. Kampen, The ins and outs of serine and glycine metabolism in cancer, Nat. Metab. 3 (2) (2021) 131-141,https://doi.org/10.1038/s42255-020-00329-9.

[8] Z. Gan, M. Zhang, D. Xie, X. Wu, C. Hong, J. Fu, L. Fan, S. Wang, S. Han, Glycinergic signaling in macrophages and its application in macrophage-associated diseases, Front. Immunol. 12 (2021), https://doi.org/10.3389/fimmu.2021.762564.

[9] S. Kang, H. Ham, C. Hong, D. Song, Glycine induces enhancement of bactericidal activity of neutrophils, Korean J. Physiol. Pharmacol. 26 (4) (2022) 229-238,https://doi.org/10.4196/kjpp.2022.26.4.229.

[10] T. Kou, J. Wu, X. Chen, Z. Chen, J. Zheng, B. Peng, Exogenous glycine promotes oxidation of glutathione and restores sensitivity of bacterial pathogens to serum-induced cell death, Redox Biol. 58 (2022) 102512, https://doi.org/10.1016/j.redox.2022.102512.

[11] O. Rom, Y. Liu, A.C. Finney, A. Ghrayeb, Y. Zhao, Y. Shukha, L. Wang, K.K. Rajanayake, S. Das, N.A. Rashdan, N. Weissman, L. Delgadillo, B. Wen, M.T. Garcia-Barrio, M. Aviram, C.G. Kevil, A. Yurdagul, C.B. Pattillo, J. Zhang, D. Sun, T. Hayek, E. Gottlieb, I. Mor, Y.E. Chen, Induction of glutathione biosynthesis by glycine-based treatment mitigates atherosclerosis, Redox Biol. 52 (2022) 102313, https://doi.org/10.1016/j.redox.2022.102313.

[12] I. Amelio, F. Cutruzzola, A. Antonov, M. Agostini, G. Melino, Serine and glycine metabolism in cancer, Trends Biochem. Sci. 39 (4) (2014) 191-198, https://doi.org/10.1016/j.tibs.2014.02.004.

[13] W.C. Zhang, N. Shyh-Chang, H. Yang, A. Rai, S. Umashankar, S. Ma, B.S. Soh, L.L. Sun, B.C. Tai, M.E. Nga, K.K. Bhakoo, S.R. Jayapal, M. Nichane, Q. Yu, D.A. Ahmed, C. Tan, W.P. Sing, J. Tam, A. Thirugananam, M.S. Noghabi, Y. Huei Pang, H.S. Ang, W. Mitchell, P. Robson, P. Kaldis, R.A. Soo, S. Swarup, E.H. Lim, B. Lim, Glycine decarboxylase activity drives non-small cell lung cancer tumorinitiating cells and tumorigenesis, Cell 148 (1) (2012) 259-272, https://doi.org/10.1016/j.cell.2011.11.050.

[14] A. Alptekin, B. Ye, Y. Yu, C.J. Poole, J. van Riggelen, Y. Zha, H. Ding, Glycine decarboxylase is a transcriptional target of MYCN required for neuroblastoma cell proliferation and tumorigenicity, Oncogene 38 (50) (2019) 7504-7520, https://doi.org/10.1038/s41388-019-0967-3.

[15] R. Liu, L. Zeng, R. Gong, F. Yuan, H. Shu, S. Li, mTORC1 activity regulates posttranslational modifications of glycine decarboxylase to modulate glycine metabolism and tumorigenesis, Nat. Commun. 12 (1) (2021), https://doi.org/10.1038/s41467-021-24321-3.

[16] M. Chen, Z. Xiao, Z. Huang, K. Xue, H. Xia, J. Zhou, D. Liao, Z. Liang, X. Xie, Q. Wei, L. Zhong, J. Yang, C. Liu, Y. Liu, S. Zhao, Glycine decarboxylase (GLDC) plays a crucial role in regulating energy metabolism, invasion, metastasis and immune escape for prostate cancer, Int. J. Biol. Sci. 19 (15) (2023) 4726-4743, https://doi.org/10.7150/ijbs.85893.

[17] H.L. Min, J. Kim, W.H. Kim, B.G. Jang, M.A. Kim, Epigenetic silencing of the putative tumor suppressor gene GLDC (Glycine dehydrogenase) in gastric carcinoma, Anticancer Res. 36 (1) (2016) 179-187.

[18] H. Zhuang, Q. Li, X. Zhang, X. Ma, Z. Wang, Y. Liu, X. Yi, R. Chen, F. Han, N. Zhang, Y. Li, Downregulation of glycine decarboxylase enhanced cofilin-mediated migration in hepatocellular carcinoma cells, Free Radical Bio. Med. 120 (2018) 1-12, https://doi.org/10.1016/j.freeradbiomed.2018.03.003.

[19] H. Zhuang, F. Wu, W. Wei, Y. Dang, B. Yang, X. Ma, F. Han, Y. Li, Glycine decarboxylase induces autophagy and is downregulated by miRNA-30d-5p in hepatocellular carcinoma, Cell Death Dis. 10 (3) (2019), https://doi.org/10.1038/s41419-019-1446-z.

[20] M. Ishaq, R. Ojha, A.P. Sharma, S.K. Singh, Autophagy in cancer: recent advances and future directions, Semin. Cancer Biol. 66 (2020) 171-181, https://doi.org/10.1016/j.semcancer.2020.03.010.

[21] N. Mizushima, M. Komatsu, Autophagy: renovation of cells and tissues, Cell 147 (4) (2011) 728-741, https://doi.org/10.1016/j.cell.2011.10.026.

[22] J. Debnath, N. Gammoh, K.M. Ryan, Autophagy and autophagy-related pathways in cancer, Nat. Rev. Mol. Cell Bio. 24 (8) (2023) 560-575, https://doi.org/10.1038/s41580-023-00585-z.

[23] J. Kim, Y.C. Kim, C. Fang, R.C. Russell, J.H. Kim, W. Fan, R. Liu, Q. Zhong, K.L. Guan, Differential regulation of distinct Vps34 complexes by AMPK in nutrient stress and autophagy, Cell 152 (1-2) (2013) 290-303, https://doi.org/10.1016/j.cell.2012.12.016.

[24] R.C. Russell, K.L. Guan, The multifaceted role of autophagy in cancer, EMBO J. 41 (13) (2022) e110031, https://doi.org/10.15252/embj.2021110031.

[25] G. Stjepanovic, S. Baskaran, M.G. Lin, J.H. Hurley, Vps 34 kinase domain dynamics regulate the autophagic PI 3-kinase complex, Mol. Cell 67 (3) (2017) 528-534.e3, https://doi.org/10.1016/j.molcel.2017.07.003.

[26] G. Wang, X. Jiang, P. Torabian, Z. Yang, Investigating autophagy and intricate cellular mechanisms in hepatocellular carcinoma: emphasis on cell death mechanism crosstalk, Cancer Lett. 588 (2024) 216744, https://doi.org/10.1016/j.canlet.2024.216744.

[27] S. Yin, W. Jin, Y. Qiu, L. Fu, T. Wang, H. Yu, Solamargine induces hepatocellular carcinoma cell apoptosis and autophagy via inhibiting LIF/miR-192-5p/CYR61/Akt signaling pathways and eliciting immunostimulatory tumor microenvironment, J. Hematol. Oncol. 15 (1) (2022) 32, https://doi.org/10.1186/s13045-022-01248-w.

[28] I. Pastushenko, C. Blanpain, EMT transition states during tumor progression and metastasis, Trends Cell Biol. 29 (3) (2019) 212-226, https://doi.org/10.1016/j.tcb.2018.12.001.

[29] G. Babaei, S.G. Aziz, N.Z.Z. Jaghi, EMT, cancer stem cells and autophagy; the three main axes of metastasis, Biomed. Pharmacother. 133 (2021) 110909, https://doi.org/10.1016/j.biopha.2020.110909.

[30] B. Levine, G. Kroemer, Biological functions of autophagy genes: a disease perspective, Cell 176 (1) (2019) 11-42, https://doi.org/10.1016/j.cell.2018.09.048.

[31] Y. Ohashi, S. Tremel, R.L. Williams, VPS 34 complexes from a structural perspective, J. Lipid Res. 60 (2) (2019) 229-241, https://doi.org/10.1194/jlr.R089490.

[32] B. Faubert, A. Solmonson, R.J. DeBerardinis, Metabolic reprogramming and cancer progression, Science 368 (6487) (2020) eaaw5473, https://doi.org/10.1126/science.aaw5473.

[33] C.J. Lee, H. Yoon, Metabolic adaptation and cellular stress response as targets for cancer therapy, World J. Mens Health 42 (1) (2024) 62-70, https://doi.org/10.5534/wjmh.230153.

[34] P.J. Kang, J. Zheng, G. Lee, D. Son, I.Y. Kim, G. Song, G. Park, S. You, Glycine decarboxylase regulates the maintenance and induction of pluripotency via metabolic control, Metab. Eng. 53 (2019) 35-47, https://doi.org/10.1016/j.ymben.2019.02.003.

[35] T. He, J. Zou, K. Sun, J. Yang, Global research status and frontiers on autophagy in hepatocellular carcinoma: a comprehensive bibliometric and visualized analysis, Int. J. Surg. 110 (5) (2024), https://doi.org/10.1097/JS9.0000000000001202.

[36] R. Rakesh, L.C. PriyaDharshini, K.M. Sakthivel, R.R. Rasmi,Role and regulation of autophagy in cancer, Biochim. Biophys. Acta Mol. Basis Dis. 1868 (7) (2022) 166400, https://doi.org/10.1016/j.bbadis.2022.166400.

[37] M.J. Czaja, W. Ding, T.M. Donohue, S.L. Friedman, J. Kim, M. Komatsu, J.J. Lemasters, A. Lemoine, J.D. Lin, J.J. Ou, D.H. Perlmutter, G. Randall, R.B. Ray, A. Tsung, X. Yin, Functions of autophagy in normal and diseased liver, Autophagy 9 (8) (2013) 1131-1158, https://doi.org/10.4161/auto.25063.

[38] D. Mukha, M. Fokra, A. Feldman, B. Sarvin, N. Sarvin, K. Nevo-Dinur, E. Besser, E. Hallo, E. Aizenshtein, Z.T. Schug, T. Shlomi, Glycine decarboxylase maintains mitochondrial protein lipoylation to support tumor growth, Cell Metab. 34 (5) (2022) 775-782.e9, https://doi.org/10.1016/j.cmet.2022.04.006.

[39] C. Qi, L. Zou, S. Wang, X. Mao, Y. Hu, J. Shi, Z. Zhang, H. Wu, Vps 34 inhibits hepatocellular carcinoma invasion by regulating endosome-lysosome trafficking via rab7-RILP and Rab11, Cancer Res. Treat. 54 (1) (2022) 182-198, https://doi.org/10.4143/crt.2020.578.

[40] J. Son, M. Kim, J.S. Lee, J.Y. Kim, E. Chun, K. Lee, Hepatitis B virus X protein promotes liver cancer progression through autophagy induction in response to TLR4 stimulation, Immune Netw. 21 (5) (2021), https://doi.org/10.4110/in.2021.21.e37.

[41] X. Li, Y. Xiao, Y. Zhu, P. Li, J. Zhou, J. Yang, Z. Chen, H. Du, H. Yu, Y. Guo, H. Bian, Z. Li, Regulation of autophagy by ST3GAL2-mediated α2-3 sialylated glycosphingolipids in hepatic encephalopathy, Int. J. Biol. Macromol. 278 (2024) 135196, https://doi.org/10.1016/j.ijbiomac.2024.135196.

[42] Y. Lee, B. Kim, H. Jang, W. Huh, Atg1-dependent phosphorylation of Vps34 is required for dynamic regulation of the phagophore assembly site and autophagy in Saccharomyces cerevisiae, Autophagy 19 (9) (2023) 2428-2442, https://doi.org/10.1080/15548627.2023.2182478.