WTAP regulates postnatal development of brown adipose tissue by stabilizing METTL3 in mice

Abstract

Brown adipocyte maturation during postnatal development is essential for brown adipose tissue (BAT) to protect animals against cold. Impaired maturation of brown adipocytes leads to cold intolerance. However, the molecular mechanisms that determine the maturation of brown adipocytes during postnatal development are not fully understood. Here, we identify Wilms’ tumor 1-associating protein (WTAP) as an essential regulator in the postnatal development and maturation of BAT. BAT-specific knockout of Wtap (Wtap-BKO) severely impairs maturation of BAT in vivo by decreasing the expression of BAT-selective genes, leading to the whitening of interscapular BAT (iBAT). Single nucleus RNA-sequencing analysis shows the dynamic changes of cell heterogeneity in iBAT of Wtap-BKO mice. Adult mice with WTAP deficiency in BAT display hypothermic and succumb to acute cold challenge. Mechanistically, WTAP deficiency decreases m⁶A mRNA modification by reducing the protein stability of METTL3. BAT-specific overexpression of Mettl3 partially rescues the phenotypes observed in Wtap-BKO mice. These data demonstrate that WTAP/METTL3 plays an essential role in iBAT postnatal development and thermogenesis.

Keywords

WTAP / BAT / m⁶A / postnatal development / METTL3 / PRDM16

References

[1] Lowell BB , Spiegelman BM . Towards a molecular understanding of adaptive thermogenesis Nature 2000; 404: 652- 60.

[2] Wu J , Boström P , Sparks Lauren M et al. Beige adipocytes are a distinct type of thermogenic fat cell in mouse and human Cell 2012; 150: 366- 76.

[3] Xue B , Rim J-S , Hogan JC et al. Genetic variability affects the development of brown adipocytes in white fat but not in interscapular brown fat J Lipid Res 2007; 48: 41- 51.

[4] Harms MJ , Ishibashi J , Wang W et al. Prdm16 is required for the maintenance of brown adipocyte identity and function in adult mice Cell Metab 2014; 19: 593- 604.

[5] Wang Y , Gao M , Zhu F et al. METTL3 is essential for postnatal development of brown adipose tissue and energy expenditure in mice Nat Commun 2020; 11: 1648.

[6] Seale P , Kajimura S , Yang W et al. Transcriptional control of brown fat determination by PRDM16 Cell Metab 2007; 6: 38- 54.

[7] Harms MJ , Lim HW , Ho Y et al. PRDM16 binds MED1 and controls chromatin architecture to determine a brown fat transcriptional program Genes Dev 2015; 29: 298- 307.

[8] Lefterova MI , Lazar MA . New developments in adipogenesis Trends Endocrinol Metab 2009; 20: 107- 14.

[9] Liu J , Yue Y , Han D et al. A METTL3-METTL14 complex mediates mammalian nuclear RNA N⁶-adenosine methylation Nat Chem Biol 2014; 10: 93- 5.

[10] Ping XL , Sun BF , Wang L et al. Mammalian WTAP is a regulatory subunit of the RNA N⁶-methyladenosine methyltransferase Cell Res 2014; 24: 177- 89.

[11] Geula S , Moshitch-Moshkovitz S , Dominissini D et al. m⁶A mRNA methylation facilitates resolution of naïve pluripotency toward differentiation Science 2015; 347: 1002- 6.

[12] Ma C , Chang M , Lv H et al. RNA m⁶A methylation participates in regulation of postnatal development of the mouse cerebellum Genome Biol 2018; 19: 68.

[13] Li X , Jiang Y , Sun X et al. METTL3 is required for maintaining β-cell function Metabolism 2021; 116: 154702.

[14] Li X , Yuan B , Lu M et al. The methyltransferase METTL3 negatively regulates nonalcoholic steatohepatitis (NASH) progression Nat Commun 2021; 12: 7213.

[15] Moindrot B , Cerase A , Coker H et al. A pooled shRNA screen identifies Rbm15, spen, and wtap as factors required for Xist RNA-mediated silencing Cell Rep 2015; 12: 562- 72.

[16] Horiuchi K , Umetani M , Minami T et al. Wilms’ tumor 1-associating protein regulates G2/M transition through stabilization of cyclin A2 mRNA Proc Natl Acad Sci USA 2006; 103: 17278- 83.

[17] Kobayashi M , Ohsugi M , Sasako T et al. The RNA methyltransferase complex of WTAP, METTL3, and METTL14 regulates mitotic clonal expansion in adipogenesis Mol Cell Biol 2018; 38: e00116- 18.

[18] Chen L , Wang X . Relationship between the genetic expression of WTAP and bladder cancer and patient prognosis Oncol Lett 2018; 16: 6966- 70.

[19] Jo HJ , Shim HE , Han ME et al. WTAP regulates migration and invasion of cholangiocarcinoma cells J Gastroenterol 2013; 48: 1271- 82.

[20] Chen Y , Peng C , Chen J et al. WTAP facilitates progression of hepatocellular carcinoma via m⁶A-HuR-dependent epigenetic silencing of ETS1 Mol Cancer 2019; 18: 127.

[21] Li BQ , Huang S , Shao QQ et al. WT1-associated protein is a novel prognostic factor in pancreatic ductal adenocarcinoma Oncol Lett 2017; 13: 2531- 8.

[22] Li X , Ding K , Li X et al. Deficiency of WTAP in hepatocytes induces lipoatrophy and non-alcoholic steatohepatitis (NASH) Nat Commun 2022; 13: 4549.

[23] Fukusumi Y , Naruse C , Asano M . Wtap is required for differentiation of endoderm and mesoderm in the mouse embryo Dev Dyn 2008; 237: 618- 29.

[24] Claflin KE , Flippo KH , Sullivan AI et al. Conditional gene targeting using UCP1-Cre mice directly targets the central nervous system beyond thermogenic adipose tissues Mol Metab 2022; 55: 101405.

[25] Li L , Li B , Li M et al. Brown adipocytes can display a mammary basal myoepithelial cell phenotype in vivo Mol Metab 2017; 6: 1198- 211.

[26] Tokuyama K , Himms-Hagen J . Brown adipose tissue thermogenesis, torpor, and obesity of glutamate-treated mice Am J Physiol Endocrinol Metab 1986; 251: E407- 15.

[27] Vijgen GHEJ , Bouvy ND , Teule GJJ et al. Brown adipose tissue in morbidly obese subjects PLoS One 2011; 6: e17247.

[28] Sun W , Modica S , Dong H et al. Plasticity and heterogeneity of thermogenic adipose tissue Nat Metab 2021; 3: 751- 61.

[29] Dominissini D , Moshitch-Moshkovitz S , Schwartz S et al. Topology of the human and mouse m⁶A RNA methylomes revealed by m⁶A-seq Nature 2012; 485: 201- 6.

[30] Wang W , Ishibashi J , Trefely S et al. A PRDM16-driven metabolic signal from adipocytes regulates precursor cell fate Cell Metab 2019; 30: 174- 89.e5.

[31] Tews D , Fischer-Posovszky P , Fromme T et al. FTO deficiency induces UCP-1 expression and mitochondrial uncoupling in adipocytes Endocrinology 2013; 154: 3141- 51.

[32] Wu R , Chen Y , Liu Y et al. m⁶A methylation promotes white-to-beige fat transition by facilitating Hif1a translation EMBO Rep 2021; 22: e52348.

[33] Ren X , Li X , Jia L et al. A small-molecule inhibitor of NF-κB-inducing kinase (NIK) protects liver from toxin-induced inflammation, oxidative stress, and injury FASEB J 2017; 31: 711- 8.

[34] Li X , Jia L , Chen X et al. Islet α-cell inflammation induced by NF-κB inducing kinase (NIK) leads to hypoglycemia, pancreatitis, growth retardation, and postnatal death in mice Theranostics 2018; 8: 5960- 71.

[35] Yao L , Cui X , Chen Q et al. Cold-inducible SIRT6 regulates thermogenesis of brown and beige fat Cell Rep 2017; 20: 641- 54.

[36] Rajakumari S , Wu J , Ishibashi J et al. EBF2 determines and maintains brown adipocyte identity Cell Metab 2013; 17: 562- 74.

[37] Hsu PJ , He C . Identifying the m⁶A methylome by affinity purification and sequencing Methods Mol Biol 2018; 1649: 49- 57.

[38] Dominissini D , Moshitch-Moshkovitz S , Salmon-Divon M et al. Transcriptome-wide mapping of N⁶-methyladenosine by m⁶A-seq based on immunocapturing and massively parallel sequencing Nat Protoc 2013; 8: 176- 89.

[39] Nagy C , Maitra M , Tanti A et al. Single-nucleus transcriptomics of the prefrontal cortex in major depressive disorder implicates oligodendrocyte precursor cells and excitatory neurons Nat Neurosci 2020; 23: 771- 81.

[40] Li J , Li E , Czepielewski RS et al. Neurotensin is an anti-thermogenic peptide produced by lymphatic endothelial cells Cell Metab 2021; 33: 1449- 65.e6.

[41] Emont MP , Jacobs C , Essene AL et al. A single-cell atlas of human and mouse white adipose tissue Nature 2022; 603: 926- 33.

[42] Zhang WC , Wells JM , Chow KH et al. miR-147b-mediated TCA cycle dysfunction and pseudohypoxia initiate drug tolerance to EGFR inhibitors in lung adenocarcinoma Nat Metab 2019; 1: 460- 74.

RIGHTS & PERMISSIONS

2022 The Author(s) 2022. Published by Oxford University Press on behalf of Higher Education Press.