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Pharmacological effect of human melanocortin-2 receptor accessory protein 2 variants on hypothalamic melanocortin receptors

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Abstract

Purpose

Melanocortin-3 receptor (MC3R), melanocortin-4 receptor (MC4R), and a recently identified melanocortin-2 receptor accessory protein 2 (MRAP2), are highly expressed in hypothalamus and coordinately regulate energy homeostasis, but the single cellular transcriptome of melanocortin system remains unknown. Several infrequent MRAP2 variants are reported from severe obese human patients but the mechanisms on how they affect melanocortin signaling are unclear.

Methods

First, we performed in silico analysis of mouse hypothalamus RNA sequencing datasets at single-cell resolution from two independent studies. Next, we inspected the three-dimensional conformational alteration of three mutations on MRAP2 protein. Finally, the influence of MRAP2 variants on MC3R and MC4R signaling was analyzed in vitro.

Results

(1) We confirmed the actual co-expression of Mrap2 with Mc3r and Mc4r, and demonstrated more broad distribution of Mrap2-positive neuronal populations than Mc3r or Mc4r in mouse hypothalamus. (2) Compared with wild-type MRAP2, MRAP2N88Y, and MRAP2R125C showed impaired α-MSH-induced MC4R or MC3R stimulation. (3) MRAP2N88Yexhibited enhanced interaction with MC4R protein and its own.

Conclusions

This is the first dedicated description of single-cell transcriptome signature of Mrap2, Mc3r, and Mc4r in the central nerve system and the first evidence describing the unique dimer formation, conformational change, and pharmacological effect of MRAP2 mutations on MC3R signaling.

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Acknowledgements

We would like to thank Xin Xie for providing the pCRE-luc plasmids. We thank Wenbiao Chen for helpful discussion and valuable comments on the manuscript preparation. The work was supported by grants from National Key Research and Development Program of China (Grant No. 2017YFA0103900, 2017YFA0103902, and 2016YFA0102200); The National Natural Science Foundation of China (Grant No. 81570760 and 31771283); One Thousand Youth Talents Program of China to Chao Zhang and Bing Luan; The Program for Professor of Special Appointment (Eastern Scholar) at Shanghai Institutions of Higher Learning (No. A11323); The Shanghai Rising-Star Program (Grant No 15QA1403600); the Fundamental Research Funds for the Central Universities of Tongji University.

Author contributions

J.L., L.L., L.G., and C.Z. conceived and designed the experiments; J.L., L.L., X.J., B.X., and L.P. performed experiments; J.L., L.L., X.J., B.X., S.L., W.Z., and C.Z. analyzed data; J.L., L.L., C.Z., B.L., L.G., and C.Z. interpreted results of experiments; J.L., L.L., and C.Z. approved final version of manuscript.

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Author notes

  1. These authors contributed equally: Jinye Liang and Lei Li.

Authors and Affiliations

  1. Translational Medical Center for Stem Cell Therapy and Institute for Regenerative Medicine, Shanghai East Hospital, Shanghai Key Laboratory of Signaling and Disease Research, School of Life Sciences and Technology, Tongji University, Shanghai, China

    Jinye Liang, Lei Li, Xuanxuan Jin, Bingxin Xu, Linyu Pi, Shangyun Liu, Wei Zhu, Cong Zhang, Lulu Gong & Chao Zhang

  2. Department of Endocrinology, Shanghai Tenth People’s Hospital, School of Medicine, Tongji University, Shanghai, China

    Bing Luan

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  1. Jinye Liang
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Liang, J., Li, L., Jin, X. et al. Pharmacological effect of human melanocortin-2 receptor accessory protein 2 variants on hypothalamic melanocortin receptors. Endocrine 61, 94–104 (2018). https://doi.org/10.1007/s12020-018-1596-2

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  • DOI: https://doi.org/10.1007/s12020-018-1596-2

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