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Genetic and functional characterization of clonally derived adult human brown adipocytes


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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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.

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Authors and Affiliations



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.

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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).

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