Nicotinamide mononucleotide protects STAT1 from oxidative stress-induced degradation to prevent colorectal tumorigenesis

Abstract

Colitis, accompanied by the accumulation of reactive oxygen species (ROS) in the intestinal tract, is a risk factor for colorectal cancer (CRC). Our previous studies indicate that nicotinamide mononucleotide (NMN) replenishment reduces chronic inflammation. In this study, we confirm that NMN supplementation reduces inflammatory cytokine levels and oxidative tissue damage in an azoxymethane/dextran sulfate sodium (AOM/DSS)-induced colitis-associated cancer (CAC) model. Mice treated with NMN developed fewer colon tumors than untreated animals under the same AOM/DSS treatment conditions. Quantitative proteomic analysis revealed a decrease in signal transducer and activator of transcription 1 (STAT1) expression in the CAC model. We demonstrate that STAT1 overexpression induces G1 arrest by downregulating CDK6 expression and suppressing tumor cell proliferation and migration. Of note, H₂O₂ induced trioxidation of the STAT1 protein and promoted its degradation, which was partially reversed by NMN supplementation. Upon H₂O₂ treatment, Cys155 in STAT1 was oxidized to sulfonic acid, whereas the mutation of Cys155 to alanine abolished ROS-mediated STAT1 degradation. These results indicate that oxidative stress induces STAT1 degradation in tumor cells and possibly in CAC tissues, whereas supplementation with NMN protects STAT1 from oxidation-induced degradation and prevents tumorigenesis. This study provides experimental evidence for the development of NMN-mediated chemoprevention strategies for CRC.

Keywords

colorectal cancer / cysteine trioxidation / nicotinamide mononucleotide / STAT1

References

[1] Sung H, Ferlay J, Siegel RL, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2021; 71: 209-249.

[2] Fujita M, Matsubara N, Matsuda I, et al. Genomic landscape of colitis-associated cancer indicates the impact of chronic inflammation and its stratification by mutations in the Wnt signaling. Oncotarget. 2017; 9: 969-981.

[3] Yu W, Tu Y, Long Z, et al. Reactive oxygen species bridge the gap between chronic inflammation and tumor development. Oxid Med Cell Long. 2022; 2022: 1-22.

[4] Jelic MD, Mandic AD, Maricic SM, Srdjenovic BU. Oxidative stress and its role in cancer. J Cancer Res Ther. 2021; 17: 22-28.

[5] Helsingen LM, Hardin CC, Kalager M. Colorectal cancer screening—approach, evidence, and future directions. NEJM Evid. 2022; 1: EVIDra210003.

[6] Lepore Signorile M, Grossi V, Fasano C, Simone C. Colorectal cancer chemoprevention: a dream coming true?. Int J Mol Sci. 2023; 24: 7597.

[7] Lv H, Lv G, Chen C, et al. NAD⁺ metabolism maintains inducible PD-L1 expression to drive tumor immune evasion. Cell Metab. 2021; 33: 110-127. e115.

[8] Guo X, Tan S, Wang T, et al. NAD⁺ salvage governs mitochondrial metabolism, invigorating natural killer cell antitumor immunity. Hepatology. 2022; 78(2): 468-485.

[9] Li X, Wang F, Xu X, Zhang J, Xu G. The dual role of STAT1 in ovarian cancer: insight into molecular mechanisms and application potentials. Front Cell Dev Biol. 2021; 9: 636595.

[10] Sun Y, Yang S, Sun N, Chen J. Differential expression of STAT1 and p21 proteins predicts pancreatic cancer progression and prognosis. Pancreas. 2014; 43: 619-623.

[11] Friedrich K, Dolznig H, Han X, Moriggl R. Steering of carcinoma progression by the YIN/YANG interaction of STAT1/STAT3. BioScience Trends. 2017; 11: 1-8.

[12] Leon-Cabrera S, Vázquez-Sandoval A, Molina-Guzman E, et al. Deficiency in STAT1 signaling predisposes gut inflammation and prompts colorectal cancer development. Cancers (Basel). 2018; 10: 341.

[13] Slattery ML, Lundgreen A, Kadlubar SA, Bondurant KL, Wolff RK. JAK/STAT/SOCS-signaling pathway and colon and rectal cancer. Mol Carcinog. 2013; 52: 155-166.

[14] Paulsen CE, Carroll KS. Cysteine-mediated redox signaling: chemistry, biology, and tools for discovery. Chem Rev. 2013; 113: 4633-4679.

[15] Wang W, Hu Y, Wang X, Wang Q, Deng H. ROS-mediated 15-hydroxyprostaglandin dehydrogenase degradation via cysteine oxidation promotes NAD+-mediated epithelial-mesenchymal transition. Cell Chem Biol. 2018; 25: 255-261.

[16] Butturini E, Carcereri de Prati A, Mariotto S. Redox regulation of STAT1 and STAT3 signaling. Int J Mol Sci. 2020; 21: 7034.

[17] Kaur N, Lu B, Monroe RK, Ward SM, Halvorsen SW. Inducers of oxidative stress block ciliary neurotrophic factor activation of Jak/STAT signaling in neurons. J Neurochem. 2005; 92: 1521-1530.

[18] Parang B, Barrett CW, Williams CS. AOM/DSS model of colitis-associated cancer. Methods Mol Biol. 2016; 1422: 297-307.

[19] Denk D, Greten FR. Inflammation: the incubator of the tumor microenvironment. Trends Cancer. 2022; 8: 901-914.

[20] Gan CS, Chong PK, Pham TK, Wright PC. Technical, experimental, and biological variations in isobaric tags for relative and absolute quantitation (iTRAQ). J Proteome Res. 2007; 6: 821-827.

[21] Zhang L, Li Q, Yang J, et al. Cytosolic TGM2 promotes malignant progression in gastric cancer by suppressing the TRIM21-mediated ubiquitination/degradation of STAT1 in a GTP binding-dependent modality. Cancer Commun. 2022; 43: 123-149.

[22] Liu M, Yang J, Xu B, Zhang X. Tumor metastasis: mechanistic insights and therapeutic interventions. MedComm. 2021; 2: 587-617.

[23] Ju H, Li X, Hong L, et al. Mediation of multiple pathways regulating cell proliferation, migration, and apoptosis in the human malignant glioma cell line U87MG via unphosphorylated STAT1: laboratory investigation. J Neurosurg. 2013; 118: 1239-1247.

[24] Delgado-Ramirez Y, Baltazar-Perez I, Martinez Y, et al. STAT1 is required for decreasing accumulation of granulocytic cells via IL-17 during initial steps of colitis-associated cancer. Int J Mol Sci. 2021; 22: 7695.

[25] Zardo G. The role of H3K4 trimethylation in CpG islands hypermethylation in cancer. Biomolecules. 2021; 11: 143.

[26] Ng J, Yu J. Promoter hypermethylation of tumour suppressor genes as potential biomarkers in colorectal cancer. Int J Mol Sci. 2015; 16: 2472-2496.

[27] Xu YP, Lv L, Liu Y, et al. Tumor suppressor TET2 promotes cancer immunity and immunotherapy efficacy. J Clin Invest. 2019; 129: 4316-4331.

[28] Goel S, Bergholz JS, Zhao JJ. Targeting CDK4 and CDK6 in cancer. Nat Rev Cancer. 2022; 22: 356-372.

[29] Dimco G, Knight RA, Latchman DS, Stephanou A. STAT1 interacts directly with cyclin D1/Cdk4 and mediates cell cycle arrest. Cell Cycle. 2010; 9: 4638-4649.

[30] Masamoto Y, Chiba A, Mizuno H, et al. EVI1 exerts distinct roles in AML via ERG and cyclin D1 promoting a chemoresistant and immune-suppressive environment. Blood Adv. 2023; 7: 1577-1593.

[31] Schmitt MJ, Philippidou D, Reinsbach SE, et al. Interferon-γ-induced activation of signal transducer and activator of transcription 1 (STAT1) up-regulates the tumor suppressing microRNA-29 family in melanoma cells. Cell Commun Signal. 2012; 10: 41.

[32] Zhang Q, Li Y, Zhao R, et al. The gain-of-function mutation E76K in SHP2 promotes CAC tumorigenesis and induces EMT via the Wnt/beta-catenin signaling pathway. Mol Carcinog. 2018; 57: 619-628.

[33] Wickstrom SA, Niessen CM. Cell adhesion and mechanics as drivers of tissue organization and differentiation: local cues for large scale organization. Curr Opin Cell Biol. 2018; 54: 89-97.

[34] Ma Y, Yi M, Wang W, et al. Oxidative degradation of dihydrofolate reductase increases CD38-mediated ferroptosis susceptibility. Cell Death Dis. 2022; 13: 944.

[35] Gu X, Nardone C, Kamitaki N, Mao A, Elledge SJ, Greenberg ME. The midnolin-proteasome pathway catches proteins for ubiquitination-independent degradation. Science. 2023; 381: eadh5021.

[36] Zhou YX, Wei J, Deng G, et al. Delivery of low-density lipoprotein from endocytic carriers to mitochondria supports steroidogenesis. Nat Cell Biol. 2023; 25: 937-949.

[37] Chen X, Vinkemeier U, Zhao Y, Jeruzalmi D, Darnell JE Jr, Kuriyan J. Crystal structure of a tyrosine phosphorylated STAT-1 dimer bound to DNA. Cell. 1998; 93: 827-839.

[38] Zhu M, John S, Berg M, Leonard WJ. Functional association of Nmi with Stat5 and Stat1 in IL-2-and IFN γ-mediated signaling. Cell. 1999; 96: 121-130.

[39] Hong W, Mo F, Zhang Z, Huang M, Wei X. Nicotinamide mononucleotide: a promising molecule for therapy of diverse diseases by targeting NAD+ metabolism. Front Cell Dev Biol. 2020; 8: 246.

[40] Zhou X, Du HH, Long X, et al. beta-nicotinamide mononucleotide (NMN) administrated by intraperitoneal injection mediates protection against UVB-induced skin damage in mice. J Inflamm Res. 2021; 14: 5165-5182.

[41] Zong Z, Liu J, Wang N, et al. Nicotinamide mononucleotide inhibits hepatic stellate cell activation to prevent liver fibrosis via promoting PGE2 degradation. Free Radic Biol Med. 2021; 162: 571-581.

[42] Takeda K, Okumura K. Nicotinamide mononucleotide augments the cytotoxic activity of natural killer cells in young and elderly mice. Biomed Res. 2021; 42: 173-179.

[43] Wada H, Otsuka R, Germeraad WTV, Murata T, Kondo T, Seino K-I. Tumor cell-induced macrophage senescence plays a pivotal role in tumor initiation followed by stable growth in immunocompetent condition. J Immunother Cancer. 2023; 11: e006677.

[44] Liu J, Zong Z, Zhang W, et al. Nicotinamide mononucleotide alleviates lps-induced inflammation and oxidative stress via decreasing COX-2 expression in macrophages. Front Mol Biosci. 2021; 8: 702107.

[45] Raynes R, Pomatto LC, Davies KJ. Degradation of oxidized proteins by the proteasome: distinguishing between the 20S, 26S, and immunoproteasome proteolytic pathways. Mol Aspects Med. 2016; 50: 41-55.

[46] Pickering AM, Davies KJA. Differential roles of proteasome and immunoproteasome regulators Pa28αβ Pa28γ and Pa200 in the degradation of oxidized proteins. Arch Biochem Biophys. 2012; 523: 181-190.

[47] Lin P-C, Lin Y-J, Lee C-T, Liu H-S, Lee J-C. Cyclooxygenase-2 expression in the tumor environment is associated with poor prognosis in colorectal cancer patients. Oncol Lett. 2013; 6: 733-739.

[48] Mostafa TM, Alm El-Din MA, Rashdan AR. Celecoxib as an adjuvant to chemotherapy for patients with metastatic colorectal cancer: a randomized controlled clinical study. Saudi Med J. 2022; 43: 37-44.

[49] Cizkova K, Foltynkova T, Gachechiladze M, Tauber Z. Comparative analysis of immunohistochemical staining intensity determined by light microscopy, ImageJ and QuPath in placental hofbauer cells. Acta Histochem Cytochem. 2021; 54: 21-29.

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