Vitamin A deficiency triggers colonic methylation potentially impairing colonic neuron via downregulation SGK1/FOXO pathway

Abstract

DNA methylation is widely involved in the modification of intestinal function, but the methylation mechanism in the enteric nervous system has not been studied in vitamin A deficiency (VAD). Herein, we firstly found that in the VAD group, gastrointestinal transit time was delayed compared with the vitamin A normal (VAN) group. RNA sequencing between VAD and VAN rats identified enriched pathways associated with enteric nerves and methylation transferase complexes. Then expression levels of DNA methyltransferases (DNMT1, DNMT3a and DNMT3b) were validated to significant increase in the VAD group. Representative reduced bisulfate sequencing showed that the VAD rats had high levels of DNA methylation in promoters and exons compared with the VAN rats. A combined methylomic and transcriptomic analysis identified that methylation levels of Sgk1, a key gene associated with enteric neural development, were elevated in the VAD group, and the activity of the SGK1/FOXO signaling axis was reduced. Furthermore, the colonic neuronal morphology and synaptic architecture were impaired in the VAD offspring. Interestingly, the above alterations in the VAD group were alleviated by vitamin A (VA) supplementation in the early postnatal period. These data suggest that VAD triggers colonic hypermethylation, which probably downregulates the SGK1/FOXO signaling pathway to cause colonic transfer dysfunction.

Keywords

enteric neurons / methylation / SGK1/FOXO pathway / vitamin A supplementation / vitamin A deficiency

References

[1] Darnton-HillI. Public health aspects in the prevention and control of vitamin deficiencies. Curr Dev Nutr. 2019; 3(9):nzz075.

[2] WisemanEM, Bar-El Dadon S, ReifenR. The vicious cycle of vitamin a deficiency: a review. Crit Rev Food Sci Nutr. 2017; 57(17):3703-3714.

[3] CantornaMT, SnyderL, AroraJ. Vitamin A and vitamin D regulate the microbial complexity, barrier function, and the mucosal immune responses to ensure intestinal homeostasis. Crit Rev Biochem Mol Biol. 2019; 54(2):184-192.

[4] XiaoL, CuiT, LiuS, et al. Vitamin A supplementation improves the intestinal mucosal barrier and facilitates the expression of tight junction proteins in rats with diarrhea. Nutrition. 2019; 57:97-108.

[5] LiY-Y, GaoY, CuiT, et al. Retinoic acid facilitates toll-like receptor 4 expression to improve intestinal barrier function through retinoic acid receptor beta. Cell Physiol Biochem. 2017; 42(4):1390-1406.

[6] LiuX, LiY-Y, WangY-T, et al. Gestational vitamin A deficiency reduces the intestinal immune response by decreasing the number of immune cells in rat offspring. Nutrition. 2014; 30(3):350-357.

[7] ChenB-L, LiuS, FengD, et al. Vitamin A deficiency in the early-life periods alters a diversity of the colonic mucosal microbiota in rats. Front Nutr. 2020; 7:580780.

[8] FrithTJR, Gogolou A, HacklandJOS, et al. Retinoic acid accelerates the specification of enteric neural progenitors from In-vitro -derived neural crest. Stem Cell Rep. 2020; 15(3):557-565.

[9] ChenJC, YangW, TsengLY, Chang HL. Enteric neural network assembly was promoted by basic fibroblast growth factor and vitamin A but inhibited by epidermal growth factor. Cells. 2022; 11(18):2841.

[10] GaoT, Wright-Jin EC, SenguptaR, AndersonJB, Heuckeroth RO. Cell-autonomous retinoic acid receptor signaling has stage-specific effects on mouse enteric nervous system. JCI Insight. 2021; 6(10):e145854.

[11] FuM, SatoY, Lyons-WarrenA, et al. Vitamin A facilitates enteric nervous system precursor migration by reducing Pten accumulation. Development. 2010; 137(4):631-640.

[12] TanM, YangT, LiuH, et al. Maternal vitamin A deficiency impairs cholinergic and nitrergic neurons, leading to gastrointestinal dysfunction in rat offspring via RARβ. Life Sci. 2021; 264:118688.

[13] JorgensenBG, BerentRM, HaSE, et al. DNA methylation, through DNMT1, has an essential role in the development of gastrointestinal smooth muscle cells and disease. Cell Death Dis. 2018; 9(5):474.

[14] LiuY-R, BaF, ChengL-J, Li X, ZhangS-W, ZhangS-C. Efficacy of Sox10 promoter methylation in the diagnosis of intestinal neuronal dysplasia from the peripheral blood. Clin Transl Gastroenterol. 2019; 10(12):e00093.

[15] WuJ-M, ZhaoY, WangX, et al. Dietary nutrients shape gut microbes and intestinal mucosa via epigenetic modifications. Crit Rev Food Sci Nutr. 2022; 62(3):783-797.

[16] LawPP, Holland ML. DNA methylation at the crossroads of gene and environment interactions. Essays Biochem. 2019; 63(6):717-726.

[17] Bar-El DadonS, ReifenR. Vitamin A and the epigenome. Crit Rev Food Sci Nutr. 2017; 57(11):2404-2411.

[18] FengY, ZhaoL-Z, HongL, Shan C, ShiW, CaiW. Alteration in methylation pattern of GATA-4 promoter region in vitamin A-deficient offspring’s heart. J Nutr Biochem. 2013; 24(7):1373-1380.

[19] ArreguínA, Ribot J, MušinovićH, von LintigJ, Palou A, BonetML. Dietary vitamin A impacts DNA methylation patterns of adipogenesis-related genes in suckling rats. Arch Biochem Biophys. 2018; 650:75-84.

[20] PassadorJ, Toffoli LV, FernandesKB, Neves-SouzaRD, PelosiGG, GomesMV. Dietary ingestion of calories and micronutrients modulates the DNA methylation profile of leukocytes from older individuals. J Nutr Health Aging. 2018; 22(10):1281-1285.

[21] AngrisanoT, Sacchetti S, NataleF, et al. Chromatin and DNA methylation dynamics during retinoic acid-induced RET gene transcriptional activation in neuroblastoma cells. Nucleic Acids Res. 2011; 39(6):1993-2006.

[22] YanJ-Y, XiaoL, FengD, et al. Vitamin A deficiency suppresses CEACAM1 to impair colonic epithelial barrier function via downregulating microbial-derived short-chain fatty acids. Genes Dis. 2023; 11(2):1066-1081.

[23] LevMG, YearimA, AstG. The alternative role of DNA methylation in splicing regulation. Trends Genet. 2015; 31(5):274-280.

[24] QuadroL, GambleMV, VogelS, et al. Retinol and retinol-binding protein: gut integrity and circulating immunoglobulins. J Infect Dis. 2000; 182(suppl 1): S97-S102.

[25] TurfkruyerM, RekimaA, MacchiaverniP, et al. Oral tolerance is inefficient in neonatal mice due to a physiological vitamin A deficiency. Mucosal Immunol. 2016; 9(2):479-491.

[26] LounderDT, Khandelwal P, DandoyCE, et al. Lower levels of vitamin A are associated with increased gastrointestinal graft-versus-host disease in children. Blood. 2017; 129(20):2801-2807.

[27] ChengB-L, ZhuJ, YangT, et al. Vitamin A deficiency exacerbates autism-like behaviors and abnormalities of the enteric nervous system in a valproic acid-induced rat model of autism. Neurotoxicology. 2020; 79:184-190.

[28] RobertsonKD. DNA methylation and human disease. Nat Rev Genet. 2005; 6(8):597-610.

[29] LiZ-F, LiY-J, HouY-J, et al. Association of plasma vitamins and carotenoids, DNA methylation of LCAT, and risk of age-related macular degeneration. Nutrients. 2023; 15(13):2985.

[30] LykoF. The DNA methyltransferase family: a versatile toolkit for epigenetic regulation. Nat Rev Genet. 2018; 19(2):81-92.

[31] HaggertyC, Kretzmer H, RiemenschneiderC, et al. Dnmt1 has de novo activity targeted to transposable elements. Nat Struct Mol Biol. 2021; 28(7):594-603.

[32] ChenT-P, UedaY, DodgeJE, Wang Z-J, LiE. Establishment and maintenance of genomic methylation patterns in mouse embryonic stem cells by Dnmt3a and Dnmt3b. Mol Cell Biol. 2003; 23(16):5594-5605.

[33] SahinP, McCaigC, JeevahanJ, Murray JT, HainsworthAH. The cell survival kinase SGK1 and its targets FOXO3a and NDRG1 in aged human brain. Neuropathol Appl Neurobiol. 2013; 39(6):623-633.

[34] BrunetA, ParkJ, TranH, Hu LS, HemmingsBA, GreenbergME. Protein kinase SGK mediates survival signals by phosphorylating the forkhead transcription factor FKHRL1 (FOXO3a). Mol Cell Biol. 2001; 21(3):952-965.

[35] GaoY-F, TangY, ZhangH-J, et al. Vincristine leads to colonic myenteric neurons injury via pro-inflammatory macrophages activation. Biochem Pharmacol. 2021; 186:114479.

[36] RoSH, LiuD, YeoH, PaikJH. FoxOs in neural stem cell fate decision. Arch Biochem Biophys. 2013; 534(1-2):55-63.

[37] KangY-N, FungC, Vanden BergheP. Gut innervation and enteric nervous system development: a spatial, temporal and molecular tour de force. Development. 2021; 148(3):dev182543.

[38] FilpaV, MoroE, ProtasoniM, Crema F, FrigoG, GiaroniC. Role of glutamatergic neurotransmission in the enteric nervous system and brain-gut axis in health and disease. Neuropharmacology. 2016; 111:14-33.

RIGHTS & PERMISSIONS

2024 The Author(s). Pediatric Discovery published by John Wiley & Sons Australia, Ltd on behalf of Children’s Hospital of Chongqing Medical University.