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Genetic and functional characterization of clonally derived adult human brown adipocytes

Abstract

Brown adipose tissue (BAT) acts in mammals as a natural defense system against hypothermia, and its activation to a state of increased energy expenditure is believed to protect against the development of obesity. Even though the existence of BAT in adult humans has been widely appreciated1,2,3,4,5,6,7,8, its cellular origin and molecular identity remain elusive largely because of high cellular heterogeneity within various adipose tissue depots. To understand the nature of adult human brown adipocytes at single cell resolution, we isolated clonally derived adipocytes from stromal vascular fractions of adult human BAT from two individuals and globally analyzed their molecular signatures. We used RNA sequencing followed by unbiased genome-wide expression analyses and found that a population of uncoupling protein 1 (UCP1)-positive human adipocytes possessed molecular signatures resembling those of a recruitable form of thermogenic adipocytes (that is, beige adipocytes). In addition, we identified molecular markers that were highly enriched in UCP1-positive human adipocytes, a set that included potassium channel K3 (KCNK3) and mitochondrial tumor suppressor 1 (MTUS1). Further, we functionally characterized these two markers using a loss-of-function approach and found that KCNK3 and MTUS1 were required for beige adipocyte differentiation and thermogenic function. The results of this study present new opportunities for human BAT research, such as facilitating cell-based disease modeling and unbiased screens for thermogenic regulators.

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Figure 1: Isolation of clonal brown adipocytes from adult human BAT.
Figure 2: Genome-wide gene expression analyses indicate a close relationship between human brown adipocytes and mouse beige adipocytes.
Figure 3: Identification of human brown adipocyte markers.
Figure 4: Mtus1 and Kcnk3 are required for beige adipocyte differentiation and thermogenic function.

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Gene Expression Omnibus

References

  1. Cypess, A.M. et al. Identification and importance of brown adipose tissue in adult humans. N. Engl. J. Med. 360, 1509–1517 (2009).

    CAS  Article  Google Scholar 

  2. Ouellet, V. et al. Outdoor temperature, age, sex, body mass index, and diabetic status determine the prevalence, mass, and glucose-uptake activity of 18F-FDG-detected BAT in humans. J. Clin. Endocrinol. Metab. 96, 192–199 (2011).

    CAS  Article  Google Scholar 

  3. van Marken Lichtenbelt, W.D. et al. Cold-activated brown adipose tissue in healthy men. N. Engl. J. Med. 360, 1500–1508 (2009).

    CAS  Article  Google Scholar 

  4. Yoneshiro, T. et al. Age-related decrease in cold-activated brown adipose tissue and accumulation of body fat in healthy humans. Obesity (Silver Spring) 19, 1755–1760 (2011).

    Article  Google Scholar 

  5. Saito, M. et al. High incidence of metabolically active brown adipose tissue in healthy adult humans: effects of cold exposure and adiposity. Diabetes 58, 1526–1531 (2009).

    CAS  Article  Google Scholar 

  6. Nedergaard, J., Bengtsson, T. & Cannon, B. Unexpected evidence for active brown adipose tissue in adult humans. Am. J. Physiol. Endocrinol. Metab. 293, E444–E452 (2007).

    CAS  Article  Google Scholar 

  7. Virtanen, K.A. et al. Functional brown adipose tissue in healthy adults. N. Engl. J. Med. 360, 1518–1525 (2009).

    CAS  Article  Google Scholar 

  8. Lidell, M.E. et al. Evidence for two types of brown adipose tissue in humans. Nat. Med. 19, 631–634 (2013).

    CAS  Article  Google Scholar 

  9. Seale, P. et al. PRDM16 controls a brown fat/skeletal muscle switch. Nature 454, 961–967 (2008).

    CAS  Article  Google Scholar 

  10. Atit, R. et al. Beta-catenin activation is necessary and sufficient to specify the dorsal dermal fate in the mouse. Dev. Biol. 296, 164–176 (2006).

    CAS  Article  Google Scholar 

  11. Timmons, J.A. et al. Myogenic gene expression signature establishes that brown and white adipocytes originate from distinct cell lineages. Proc. Natl. Acad. Sci. USA 104, 4401–4406 (2007).

    CAS  Article  Google Scholar 

  12. Ohno, H., Shinoda, K., Ohyama, K., Sharp, L.Z. & Kajimura, S. EHMT1 controls brown adipose cell fate and thermogenesis through the PRDM16 complex. Nature 504, 163–167 (2013).

    CAS  Article  Google Scholar 

  13. Harms, M. & Seale, P. Brown and beige fat: development, function and therapeutic potential. Nat. Med. 19, 1252–1263 (2013).

    CAS  Article  Google Scholar 

  14. Kajimura, S. & Saito, M. A new era in brown adipose tissue biology: molecular control of brown fat development and energy homeostasis. Annu. Rev. Physiol. 76, 225–249 (2014).

    CAS  Article  Google Scholar 

  15. Waldén, T.B., Hansen, I.R., Timmons, J.A., Cannon, B. & Nedergaard, J. Recruited vs. nonrecruited molecular signatures of brown, “brite,” and white adipose tissues. Am. J. Physiol. Endocrinol. Metab. 302, E19–E31 (2012).

    Article  Google Scholar 

  16. Wu, J. et al. Beige adipocytes are a distinct type of thermogenic fat cell in mouse and human. Cell 150, 366–376 (2012).

    CAS  Article  Google Scholar 

  17. Ohno, H., Shinoda, K., Spiegelman, B.M. & Kajimura, S. PPARgamma agonists induce a white-to-brown fat conversion through stabilization of PRDM16 protein. Cell Metab. 15, 395–404 (2012).

    CAS  Article  Google Scholar 

  18. Sharp, L.Z. et al. Human BAT possesses molecular signatures that resemble beige/brite cells. PLoS ONE 7, e49452 (2012).

    CAS  Article  Google Scholar 

  19. Lee, P., Werner, C.D., Kebebew, E. & Celi, F.S. Functional thermogenic beige adipogenesis is inducible in human neck fat. Int. J. Obes. (Lond). 38, 170–176 (2014).

    Article  Google Scholar 

  20. Jespersen, N.Z. et al. A classical brown adipose tissue mRNA signature partly overlaps with brite in the supraclavicular region of adult humans. Cell Metab. 17, 798–805 (2013).

    CAS  Article  Google Scholar 

  21. Cypess, A.M. et al. Anatomical localization, gene expression profiling and functional characterization of adult human neck brown fat. Nat. Med. 19, 635–639 (2013).

    CAS  Article  Google Scholar 

  22. Villarroya, F. & Vidal-Puig, A. Beyond the sympathetic tone: the new brown fat activators. Cell Metab. 17, 638–643 (2013).

    CAS  Article  Google Scholar 

  23. Tews, D. et al. Comparative gene array analysis of progenitor cells from human paired deep neck and subcutaneous adipose tissue. Mol. Cell. Endocrinol. 395, 41–50 (2014).

    CAS  Article  Google Scholar 

  24. Long, J.Z. et al. A smooth muscle-like origin for beige adipocytes. Cell Metab. 19, 810–820 (2014).

    CAS  Article  Google Scholar 

  25. Rajakumari, S. et al. EBF2 determines and maintains brown adipocyte identity. Cell Metab. 17, 562–574 (2013).

    CAS  Article  Google Scholar 

  26. Schulz, T.J. et al. Brown-fat paucity due to impaired BMP signalling induces compensatory browning of white fat. Nature 495, 379–383 (2013).

    CAS  Article  Google Scholar 

  27. Svensson, P.A. et al. Gene expression in human brown adipose tissue. Int. J. Mol. Med. 27, 227–232 (2011).

    CAS  Article  Google Scholar 

  28. Søndergaard, E. et al. Chronic adrenergic stimulation induces brown adipose tissue differentiation in visceral adipose tissue. Diabet. Med. 32, e4–e8 (2015).

    Article  Google Scholar 

  29. Nouet, S. et al. Trans-inactivation of receptor tyrosine kinases by novel angiotensin II AT2 receptor-interacting protein, ATIP. J. Biol. Chem. 279, 28989–28997 (2004).

    CAS  Article  Google Scholar 

  30. Chondronikola, M. et al. Brown adipose tissue improves whole-body glucose homeostasis and insulin sensitivity in humans. Diabetes 63, 4089–4099 (2014).

    CAS  Article  Google Scholar 

  31. van der Lans, A.A. et al. Cold acclimation recruits human brown fat and increases nonshivering thermogenesis. J. Clin. Invest. 123, 3395–3403 (2013).

    CAS  Article  Google Scholar 

  32. Yoneshiro, T. et al. Recruited brown adipose tissue as an antiobesity agent in humans. J. Clin. Invest. 123, 3404–3408 (2013).

    CAS  Article  Google Scholar 

  33. Lee, P. et al. Temperature-acclimated brown adipose tissue modulates insulin sensitivity in humans. Diabetes 63, 3686–3698 (2014).

    CAS  Article  Google Scholar 

  34. Trapnell, C. et al. Differential analysis of gene regulation at transcript resolution with RNA-seq. Nat. Biotechnol. 31, 46–53 (2013).

    CAS  Article  Google Scholar 

  35. Irizarry, R.A. et al. Exploration, normalization, and summaries of high density oligonucleotide array probe level data. Biostatistics 4, 249–264 (2003).

    Article  Google Scholar 

  36. Li, C. & Wong, W.H. Model-based analysis of oligonucleotide arrays: expression index computation and outlier detection. Proc. Natl. Acad. Sci. USA 98, 31–36 (2001).

    CAS  Article  Google Scholar 

  37. Saeed, A.I. et al. TM4: a free, open-source system for microarray data management and analysis. Biotechniques 34, 374–378 (2003).

    CAS  Article  Google Scholar 

  38. Stöver, B.C. & Muller, K.F. TreeGraph 2: combining and visualizing evidence from different phylogenetic analyses. BMC Bioinformatics 11, 7 (2010).

    Article  Google Scholar 

  39. Huang, W., Sherman, B.T. & Lempicki, R.A. Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat. Protoc. 4, 44–57 (2009).

    CAS  Google Scholar 

  40. Inoue, M., Harada, K., Matsuoka, H., Sata, T. & Warashina, A. Inhibition of TASK1-like channels by muscarinic receptor stimulation in rat adrenal medullary cells. J. Neurochem. 106, 1804–1814 (2008).

    CAS  PubMed  Google Scholar 

  41. Cohen, J. Statistical Power Analysis for the Behavioral Sciences (L. Erlbaum Associates, Hillsdale, N.J., 1988).

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Acknowledgements

We acknowledge support from the National Institutes of Health (NIH) (DK087853 and DK097441), the UCSF Diabetes Research Center grant (DK63720), the UCSF Program for Breakthrough Biomedical Research program, the Pew Charitable Trust, and the Japan Science and Technology Agency (all to S.K.); and from the NIH (P50-GM60338) and the American Dental Association (1-14-TS-35) (both to L.S.S.). K.S. is supported by a fellowship from the Japan Society for the Promotion of Science. Y.H. is supported by the Manpei Suzuki Diabetes Foundation. I.H.N.L. is supported by the Dutch Heart Foundation.

Author information

Authors and Affiliations

Authors

Contributions

K.S. and S.K. designed the experiments. K.S., I.H.N.L., Y.H., H.H., S.B.S., M.K. and S.K. performed the cellular experiments and analyzed the data. M.C., A.M.C., L.S.S. and Y.-H.T. provided adipose tissue samples. K.S., Y.H., H.H., R.X. and S.K. analyzed adipose tissue samples. K.S. and S.K. wrote the manuscript. All authors contributed to editing the manuscript. S.K. conceived and managed the project.

Corresponding author

Correspondence to Shingo Kajimura.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–5 (PDF 864 kb)

Supplementary Table 1

Human brown adipocyte-enriched genes in a differentiated state. (PDF 1206 kb)

Supplementary Table 2

Human brown adipocyte-enriched genes in an undifferentiated state (PDF 1153 kb)

Supplementary Table 3

Expression level of smooth muscle lineage-selective genes in human brown and white adipocytes at undifferentiated and differentiated states. (PDF 79 kb)

Supplementary Table 4

List of core brown fat-selective genes conserved in mice and humans (group A). (PDF 125 kb)

Supplementary Table 5

Primer sequences used for quantitative RT-PCR. (PDF 115 kb)

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Shinoda, K., Luijten, I., Hasegawa, Y. et al. Genetic and functional characterization of clonally derived adult human brown adipocytes. Nat Med 21, 389–394 (2015). https://doi.org/10.1038/nm.3819

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  • DOI: https://doi.org/10.1038/nm.3819

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