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Screening of potential adipokines identifies S100A4 as a marker of pernicious adipose tissue and insulin resistance

Abstract

Background

Adipokines are peptides secreted from white adipose tissue (WAT), which have been linked to WAT dysfunction and metabolic complications of obesity. We set out to identify novel adipokines in subcutaneous WAT (sWAT) linked to insulin resistance (IR).

Methods

Gene expression was determined by microarray and qPCR in obese and non-obese subjects with varying degree of IR. WAT-secreted and circulating protein levels were measured by ELISA.

Results

In sWAT of 80 obese women discordant for IR, 44 genes encoding potential adipose-secreted proteins were differentially expressed. Among these, merely two proteins, S100A4 and MXRA5 were released from sWAT in a time-dependent manner (criterion for true adipokines) but only the circulating levels of S100A4 were higher in IR. In two additional cohorts (n = 29 and n = 56), sWAT S100A4 secretion was positively and BMI-independently associated with IR (determined by clamp or HOMA-IR), ATP-III risk score and adipocyte size (hypertrophy). In non-obese (n = 20) and obese subjects before and after bariatric surgery (n = 21), circulating and sWAT-secreted levels were highest in the obese and normalized following weight loss. Serum S100A4 concentrations were higher in subjects with type 2 diabetes. S100A4 sWAT expression associated positively with genes involved in inflammation/extracellular matrix formation and inversely with genes in metabolic pathways. Although S100A4 was expressed in both stromal cells and adipocytes, only the expression in adipocytes associated with BMI.

Conclusions

S100A4 is a novel adipokine associated with IR and sWAT inflammation/adipocyte hypertrophy independently of BMI. Its value as a circulating marker for dysfunctional WAT and IR needs to be validated in larger cohorts.

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References

  1. Rosen ED, Spiegelman BM. What we talk about when we talk about fat. Cell. 2014;156:20–44.

    Article  CAS  Google Scholar 

  2. Leal Vde O, Mafra D. Adipokines in obesity. Clin Chim Acta. 2013;419:87–94.

    Article  Google Scholar 

  3. Kim J, Choi YS, Lim S, Yea K, Yoon JH, Jun DJ, et al. Comparative analysis of the secretory proteome of human adipose stromal vascular fraction cells during adipogenesis. Proteomics. 2010;10:394–405.

    Article  CAS  Google Scholar 

  4. Lamers D, Famulla S, Wronkowitz N, Hartwig S, Lehr S, Ouwens DM, et al. Dipeptidyl peptidase 4 is a novel adipokine potentially linking obesity to the metabolic syndrome. Diabetes. 2011;60:1917–25.

    Article  CAS  Google Scholar 

  5. Rosenow A, Arrey TN, Bouwman FG, Noben JP, Wabitsch M, Mariman EC, et al. Identification of novel human adipocyte secreted proteins by using SGBS cells. J Proteome Res. 2010;9:5389–401.

    Article  CAS  Google Scholar 

  6. Zhong J, Krawczyk SA, Chaerkady R, Huang H, Goel R, Bader JS, et al. Temporal profiling of the secretome during adipogenesis in humans. J Proteome Res. 2010;9:5228–38.

    Article  CAS  Google Scholar 

  7. Zvonic S, Lefevre M, Kilroy G, Floyd ZE, DeLany JP, Kheterpal I, et al. Secretome of primary cultures of human adipose-derived stem cells: modulation of serpins by adipogenesis. Mol Cell Proteom. 2007;6:18–28.

    Article  CAS  Google Scholar 

  8. Hotamisligil GS, Arner P, Caro JF, Atkinson RL, Spiegelman BM. Increased adipose tissue expression of tumor necrosis factor-alpha in human obesity and insulin resistance. J Clin Invest. 1995;95:2409–15.

    Article  CAS  Google Scholar 

  9. Lonnqvist F, Nordfors L, Jansson M, Thorne A, Schalling M, Arner P. Leptin secretion from adipose tissue in women. Relationship to plasma levels and gene expression. J Clin Invest. 1997;99:2398–404.

    Article  CAS  Google Scholar 

  10. Eto H, Suga H, Matsumoto D, Inoue K, Aoi N, Kato H, et al. Characterization of structure and cellular components of aspirated and excised adipose tissue. Plast Reconstr Surg. 2009;124:1087–97.

    Article  CAS  Google Scholar 

  11. Berry R, Jeffery E, Rodeheffer MS. Weighing in on adipocyte precursors. Cell Metab. 2014;19:8–20.

    Article  CAS  Google Scholar 

  12. Bluher M. Are there still healthy obese patients? Curr Opin Endocrinol Diabetes Obes. 2012;19:341–6.

    Article  Google Scholar 

  13. Arner P, Sahlqvist AS, Sinha I, Xu H, Yao X, Waterworth D, et al. The epigenetic signature of systemic insulin resistance in obese women. Diabetologia. 2016;59:2393–405.

    Article  CAS  Google Scholar 

  14. Eriksson Hogling D, Petrus P, Gao H, Backdahl J, Dahlman I, Laurencikiene J, et al. Adipose and circulating CCL18 levels associate with metabolic risk factors in women. J Clin Endocrinol Metab. 2016;101:4021–9. jc20162390

    Article  Google Scholar 

  15. Arner E, Mejhert N, Kulyte A, Balwierz PJ, Pachkov M, Cormont M, et al. Adipose tissue microRNAs as regulators of CCL2 production in human obesity. Diabetes. 2012;61:1986–93.

    Article  CAS  Google Scholar 

  16. Acosta JR, Douagi I, Andersson DP, Backdahl J, Ryden M, Arner P, et al. Increased fat cell size: a major phenotype of subcutaneous white adipose tissue in non-obese individuals with type 2 diabetes. Diabetologia. 2016;59:560–70.

    Article  CAS  Google Scholar 

  17. Ryden M, Andersson DP, Bergstrom IB, Arner P. Adipose tissue and metabolic alterations: regional differences in fat cell size and number matter, but differently: a cross-sectional study. J Clin Endocrinol Metab. 2014;99:E1870–6.

    Article  CAS  Google Scholar 

  18. Matthews DR, Hosker JP, Rudenski AS, Naylor BA, Treacher DF, Turner RC. Homeostasis model assessment: insulin resistance and beta-cell function from fasting plasma glucose and insulin concentrations in man. Diabetologia. 1985;28:412–9.

    Article  CAS  Google Scholar 

  19. Third Report of the National Cholesterol Education Program (NCEP) expert panel on detection, evaluation, and treatment of high blood cholesterol in adults (adult treatment panel III) final report. Circulation. 2002;106:3143–421.

  20. Gao H, Mejhert N, Fretz JA, Arner E, Lorente-Cebrian S, Ehrlund A, et al. Early B cell factor 1 regulates adipocyte morphology and lipolysis in white adipose tissue. Cell Metab. 2014;19:981–92.

    Article  CAS  Google Scholar 

  21. Hoffstedt J, Arvidsson E, Sjolin E, Wahlen K, Arner P. Adipose tissue adiponectin production and adiponectin serum concentration in human obesity and insulin resistance. J Clin Endocrinol Metab. 2004;89:1391–6.

    Article  CAS  Google Scholar 

  22. Lonnqvist F, Nordfors L, Jansson M, Thorne A, Schalling M, Arner P. Leptin secretion from adipose tissue in women. Relationship to plasma levels and gene expression. J Clin Invest. 1997;99:2398–404.

    Article  CAS  Google Scholar 

  23. Hotamisligil GS, Arner P, Caro JF, Atkinson RL, Spiegelman BM. Increased adipose tissue expression of tumor necrosis factor-alpha in human obesity and insulin resistance. J Clin Invest. 1995;95:2409–15.

    Article  CAS  Google Scholar 

  24. Fain JN. Release of inflammatory mediators by human adipose tissue is enhanced in obesity and primarily by the nonfat cells: a review. Mediat Inflamm. 2010;2010:513948.

    Article  Google Scholar 

  25. Esteve D, Boulet N, Volat F, Zakaroff-Girard A, Ledoux S, Coupaye M, et al. Human white and brite adipogenesis is supported by MSCA1 and is impaired by immune cells. Stem Cells. 2015;33:1277–91.

    Article  CAS  Google Scholar 

  26. Lehr S, Hartwig S, Lamers D, Famulla S, Muller S, Hanisch FG, et al. Identification and validation of novel adipokines released from primary human adipocytes. Mol Cell Proteom. 2012;11:M111 010504.

    Article  Google Scholar 

  27. Rosenow A, Noben JP, Bouwman FG, Mariman EC, Renes J. Hypoxia-mimetic effects in the secretome of human preadipocytes and adipocytes. Biochim Biophys Acta. 2013;1834:2761–71.

    Article  CAS  Google Scholar 

  28. Lim JM, Wollaston-Hayden EE, Teo CF, Hausman D, Wells L. Quantitative secretome and glycome of primary human adipocytes during insulin resistance. Clin Proteom. 2014;11:20.

    Article  Google Scholar 

  29. Kuk JL, Ardern CI. Are metabolically normal but obese individuals at lower risk for all-cause mortality? Diabetes Care. 2009;32:2297–9.

    Article  Google Scholar 

  30. Gross SR, Sin CG, Barraclough R, Rudland PS. Joining S100 proteins and migration: for better or for worse, in sickness and in health. Cell Mol life Sci: CMLS. 2014;71:1551–79.

    Article  CAS  Google Scholar 

  31. Malashkevich VN, Varney KM, Garrett SC, Wilder PT, Knight D, Charpentier TH, et al. Structure of Ca2+-bound S100A4 and its interaction with peptides derived from nonmuscle myosin-IIA. Biochemistry. 2008;47:5111–26.

    Article  CAS  Google Scholar 

  32. Davies M, Harris S, Rudland P, Barraclough R. Expression of the rat, S-100-related, calcium-binding protein gene, p9Ka, in transgenic mice demonstrates different patterns of expression between these two species. DNA Cell Biol. 1995;14:825–32.

    Article  CAS  Google Scholar 

  33. Naaman C EL, Grum-Schwensen B, Mansouri A, Grigorian M, Santoni-Rugiu E, Hansen T. et al. Cancer predisposition in mice deficient for the metastasis-associated Mts1(S100A4) gene. Oncogene. 2004;23:3670–80.

    Article  Google Scholar 

  34. Davies MP, Rudland PS, Robertson L, Parry EW, Jolicoeur P, Barraclough R. Expression of the calcium-binding protein S100A4 (p9Ka) in MMTV-neu transgenic mice induces metastasis of mammary tumours. Oncogene. 1996;13:1631–7.

    CAS  PubMed  Google Scholar 

  35. Hapangama DK, Raju RS, Valentijn AJ, Barraclough D, Hart A, Turner MA, et al. Aberrant expression of metastasis-inducing proteins in ectopic and matched eutopic endometrium of women with endometriosis: implications for the pathogenesis of endometriosis. Human Reprod. 2012;27:394–407.

    Article  CAS  Google Scholar 

  36. Takenaga K, Nakamura Y, Sakiyama S. Cellular localization of pEL98 protein, an S100-related calcium binding protein, in fibroblasts and its tissue distribution analyzed by monoclonal antibodies. Cell Struct Funct. 1994;19:133–41.

    Article  CAS  Google Scholar 

  37. Flynn AM, Rudland PS, Barraclough R. Protein interactions between S100A4 (p9Ka) and other cellular proteins identified using in vitro methods. Biochem Soc Trans. 1996;24:341S.

    Article  CAS  Google Scholar 

  38. Ford HL, Silver DL, Kachar B, Sellers JR, Zain SB. Effect of Mts1 on the structure and activity of nonmuscle myosin II. Biochemistry. 1997;36:16321–7.

    Article  CAS  Google Scholar 

  39. Schmidt-Hansen B, Ornas D, Grigorian M, Klingelhofer J, Tulchinsky E, Lukanidin E, et al. Extracellular S100A4(mts1) stimulates invasive growth of mouse endothelial cells and modulates MMP-13 matrix metalloproteinase activity. Oncogene. 2004;23:5487–95.

    Article  CAS  Google Scholar 

  40. Andersen K, Mori H, Fata J, Bascom J, Oyjord T, Maelandsmo GM, et al. The metastasis-promoting protein S100A4 regulates mammary branching morphogenesis. Dev Biol. 2011;352:181–90.

    Article  CAS  Google Scholar 

  41. Donato R, Cannon BR, Sorci G, Riuzzi F, Hsu K, Weber DJ, et al. Functions of S100 proteins. Curr Mol Med. 2013;13:24–57.

    Article  CAS  Google Scholar 

  42. Bhaskaran K, Douglas I, Forbes H, dos-Santos-Silva I, Leon DA, Smeeth L. Body-mass index and risk of 22 specific cancers: a population-based cohort study of 5.24 million UK adults. Lancet. 2014;384:755–65.

    Article  Google Scholar 

  43. Laforest S, Labrecque J, Michaud A, Cianflone K, Tchernof A. Adipocyte size as a determinant of metabolic disease and adipose tissue dysfunction. Crit Rev Clin Lab Sci. 2015;52:301–13.

    Article  CAS  Google Scholar 

  44. Arner E, Westermark PO, Spalding KL, Britton T, Ryden M, Frisen J, et al. Adipocyte turnover: relevance to human adipose tissue morphology. Diabetes. 2010;59:105–9.

    Article  CAS  Google Scholar 

  45. Stein U, Arlt F, Walther W, Smith J, Waldman T, Harris ED, et al. The metastasis-associated gene S100A4 is a novel target of beta-catenin/T-cell factor signaling in colon cancer. Gastroenterology. 2006;131:1486–1500.

    Article  CAS  Google Scholar 

  46. Stein U, Arlt F, Smith J, Sack U, Herrmann P, Walther W, et al. Intervening in beta-catenin signaling by sulindac inhibits S100A4-dependent colon cancer metastasis. Neoplasia. 2011;13:131–44.

    Article  CAS  Google Scholar 

  47. Aguilar-Morante D, Morales-Garcia JA, Santos A, Perez-Castillo A. CCAAT/enhancer binding protein beta induces motility and invasion of glioblastoma cells through transcriptional regulation of the calcium binding protein S100A4. Oncotarget. 2015;6:4369–84.

    Article  Google Scholar 

  48. Liu J, Xu ZM, Qiu GB, Zheng ZH, Sun KL, Fu WN. S100A4 is upregulated via the binding of c-Myb in methylation-free laryngeal cancer cells. Oncol Rep. 2014;31:442–9.

    Article  CAS  Google Scholar 

  49. Xu X, Su B, Xie C, Wei S, Zhou Y, Liu H, et al. Sonic hedgehog-Gli1 signaling pathway regulates the epithelial mesenchymal transition (EMT) by mediating a new target gene, S100A4, in pancreatic cancer cells. PLoS ONE. 2014;9:e96441.

    Article  Google Scholar 

  50. Bloor ID, Symonds ME. Sexual dimorphism in white and brown adipose tissue with obesity and inflammation. Horm Behav. 2014;66:95–103.

    Article  Google Scholar 

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Acknowledgements

We would like to thank Ms Kerstin Wåhlén for skilled technical assistance and research nurses Katarina Hertel and Yvonne Widlund for excellent help with the clinical examinations. MR, ID, and PA conceived the study. MR and PA collected all the data, EN and AT recruited all subjects, MR wrote the first version of the manuscript and is the guarantor of the data. MR, PA, PP, DE, ID and AB generated data and prepared figures/tables. MR, ID, AE, DE, PP and PA analyzed data. All authors read and contributed to the final version of the manuscript.

Funding

This work was supported by grants from the EU Innovative Medicines Initiative EMIF, the Swedish Research Council, The Swedish Diabetes Foundation, CIMED, the Diabetes Theme Center at Karolinska Institutet, the Stockholm County Council, the Erling-Persson Family Foundation and the Novo Nordisk Foundation including the Tripartite Immuno-metabolism Consortium (TrIC), Grant Number NNF15CC0018486 and the MSAM Consortium NNF15SA0018346.

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Correspondence to Mikael Rydén.

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Arner, P., Petrus, P., Esteve, D. et al. Screening of potential adipokines identifies S100A4 as a marker of pernicious adipose tissue and insulin resistance. Int J Obes 42, 2047–2056 (2018). https://doi.org/10.1038/s41366-018-0018-0

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