Abstract

Menopause is associated with dyslipidemia and an increased risk of cardio-cerebrovascular disease. The classic view assumes that the underlying mechanism of dyslipidemia is attributed to an insufficiency of estrogen. In addition to a decrease in estrogen, circulating follicle-stimulating hormone (FSH) levels become elevated at menopause. In this study, we find that blocking FSH reduces serum cholesterol via inhibiting hepatic cholesterol biosynthesis. First, epidemiological results show that the serum FSH levels are positively correlated with the serum total cholesterol levels, even after adjustment by considering the effects of serum estrogen. In addition, the prevalence of hypercholesterolemia is significantly higher in peri-menopausal women than that in pre-menopausal women. Furthermore, we generated a mouse model of FSH elevation by intraperitoneally injecting exogenous FSH into ovariectomized (OVX) mice, in which a normal level of estrogen (E2) was maintained by exogenous supplementation. Consistently, the results indicate that FSH, independent of estrogen, increases the serum cholesterol level in this mouse model. Moreover, blocking FSH signaling by anti-FSHβ antibody or ablating the FSH receptor (FSHR) gene could effectively prevent hypercholesterolemia induced by FSH injection or high-cholesterol diet feeding. Mechanistically, FSH, via binding to hepatic FSHRs, activates the Gi2α/β-arrestin-2/Akt pathway and subsequently inhibits the binding of FoxO1 with the SREBP-2 promoter, thus preventing FoxO1 from repressing SREBP-2 gene transcription. This effect, in turn, results in the upregulation of SREBP-2, which drives HMGCR nascent transcription and de novo cholesterol biosynthesis, leading to the increase of cholesterol accumulation. This study uncovers that blocking FSH signaling might be a new strategy for treating hypercholesterolemia during menopause, particularly for women in peri-menopause characterized by FSH elevation only.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    Randolph, J. J. F. et al. Change in follicle-stimulating hormone and estradiol across the menopausal transition: effect of age at the final menstrual period. J. Clin. Endocrinol. Metab. 96, 746–754 (2011).

  2. 2.

    Yang, W. et al. Serum lipids and lipoproteins in Chinese men and women. Circulation 125, 2212–2221 (2012).

  3. 3.

    Wenger, N. K. Clinical characteristics of coronary heart disease in women: emphasis on gender differences. Cardiovasc. Res. 53, 558–567 (2002).

  4. 4.

    van Beresteijn, E. C. H. et al. Perimenopausal increase in serum cholesterol: a 10-year longitudinal study. Am. J. Epidemiol. 137, 383–392 (1993).

  5. 5.

    Rossouw, J. E. et al. Risks and benefits of estrogen plus progestin in healthy postmenopausal women: principal results from the Women’s Health Initiative randomized controlled trial. JAMA 288, 321–333 (2002).

  6. 6.

    Randolph, J. F. et al. The value of follicle-stimulating hormone concentration and clinical findings as markers of the late menopausal transition. J. Clin. Endocrinol. Metab. 91, 3034–3040 (2006).

  7. 7.

    Matthews, K. A. et al. Are changes in cardiovascular disease risk factors in midlife women due to chronological aging or to the menopausal transition? J. Am. Coll. Cardiol. 54, 2366–2373 (2009).

  8. 8.

    Zhou, J. L. et al. Serum lipid profile changes during the menopausal transition in Chinese women: a community-based cohort study. Menopause 17, 997–1003 (2010).

  9. 9.

    Randolph, J. J. F. et al. Change in estradiol and follicle-stimulating hormone across the early menopausal transition: effects of ethnicity and age. J. Clin. Endocrinol. Metab. 89, 1555–1561 (2004).

  10. 10.

    Green, J. S. et al. The effects of exercise training on abdominal visceral fat, body composition, and indicators of the metabolic syndrome in postmenopausal women with and without estrogen replacement therapy: The HERITAGE family study. Metabolism 53, 1192–1196 (2004).

  11. 11.

    Song, Y. et al. Follicle-stimulating hormone induces postmenopausal dyslipidemia through inhibiting hepatic cholesterol metabolism. J. Clin. Endocrinol. Metab. 101, 254–263 (2016).

  12. 12.

    Warner, M. & Gustafsson, J.-A. On estrogen, cholesterol metabolism, and breast cancer. N. Engl. J. Med. 370, 572–573 (2014).

  13. 13.

    Manson, J. E. et al. Estrogen plus progestin and the risk of coronary heart disease. N. Engl. J. Med. 349, 523–534 (2003).

  14. 14.

    Sairam, M. R. in Encyclopedia of Reproduction, (eds Knobiland, E. & Niell, J. D.) 552–565 (Academic Press, Inc, New York, 1999).

  15. 15.

    Sowers, M. R. et al. Amount of bone loss in relation to time around the final menstrual period and follicle-stimulating hormone staging of the transmenopause. J. Clin. Endocrinol. Metab. 95, 2155–2162 (2010).

  16. 16.

    Sun, L. et al. FSH directly regulates bone mass. Cell 125, 247–260 (2006).

  17. 17.

    Liu, P. et al. Blocking FSH induces thermogenic adipose tissue and reduces body fat. Nature 546, 107 (2017).

  18. 18.

    Zhu, L. L. et al. Blocking antibody to the β-subunit of FSH prevents bone loss by inhibiting bone resorption and stimulating bone synthesis. Proc. Natl Acad. Sci. 109, 14574–14579 (2012).

  19. 19.

    Chu, M. C., Rath, K. M., Huie, J. & Taylor, H. S. Elevated basal FSH in normal cycling women is associated with unfavourable lipid levels and increased cardiovascular risk. Hum. Reprod. 18, 1570–1573 (2003).

  20. 20.

    Conti, M. Specificity of the cyclic adenosine 3′,5′-monophosphate signal in granulosa cell function. Biol. Reprod. 67, 1653–1661 (2002).

  21. 21.

    Liu, X. M. et al. FSH regulates fat accumulation and redistribution in aging through the Gαi/Ca2+/CREB pathway. Aging Cell 14, 409–420 (2015).

  22. 22.

    Qi, S. et al. HSP27 phosphorylation modulates TRAIL-induced activation of Src-Akt/ERK signaling through interaction with β-arrestin2. Cell. Signal. 26, 594–602 (2014).

  23. 23.

    min, H. K. et al. Increased hepatic synthesis and dysregulation of cholesterol metabolism is associated with the severity of nonalcoholic fatty liver disease. Cell. Metab. 15, 665–674 (2012).

  24. 24.

    Bloch, K. The biological synthesis of cholesterol. Science 150, 19–28 (1965).

  25. 25.

    Tian, L. et al. A novel role for thyroid-stimulating hormone: up-regulation of hepatic 3-hydroxy-3-methyl-glutaryl-coenzyme a reductase expression through the cyclic adenosine monophosphate/protein kinase A/cyclic adenosine monophosphate–responsive element binding protein pathway. Hepatology 52, 1401–1409 (2010).

  26. 26.

    Pertusa, M., Morenilla-Palao, C., Carteron, C., Viana, F. & Cabedo, H. Transcriptional control of cholesterol biosynthesis in schwann cells by axonal neuregulin 1. J. Biol. Chem. 282, 28768–28778 (2007).

  27. 27.

    Horton, J. D., Goldstein, J. L. & Brown, M. S. SREBPs: activators of the complete program of cholesterol and fatty acid synthesis in the liver. J. Clin. Invest. 109, 1125–1131 (2002).

  28. 28.

    Faulds, M. H., Zhao, C., Wright, K. D. & Gustafsson, J.-Å. The diversity of sex steroid action: regulation of metabolism by estrogen signaling. J. Endocrinol. 212, 3–12 (2012).

  29. 29.

    Demeestere, I. et al. Gonadotropin-releasing hormone agonist for the prevention of chemotherapy-induced ovarian failure in patients with lymphoma: 1-year follow-up of a prospective randomized trial. J. Clin. Oncol. 31, 903–909 (2013).

  30. 30.

    Jiang, X., Dias, J. A. & He, X. Structural biology of glycoprotein hormones and their receptors: insights to signaling. Mol. Cell. Endocrinol. 382, 424–451 (2014).

  31. 31.

    Jiang, X. et al. Structure of follicle-stimulating hormone in complex with the entire ectodomain of its receptor. Proc. Natl Acad. Sci. 109, 12491–12496 (2012).

  32. 32.

    Zhu, L. L. et al. Blocking FSH action attenuates osteoclastogenesis. Biochem. Biophys. Res. Commun. 422, 54–58 (2012).

  33. 33.

    Scheja, L. & Heeren, J. Metabolic interplay between white, beige, brown adipocytes and the liver. J. Hepatol. 64, 1176–1186 (2016).

  34. 34.

    Geelen, M. H., Gibson, D. & Rodwell, V. Hydroxymethylglutaryl-CoA reductase—the rate-limiting enzyme of cholesterol biosynthesis. FEBS Lett. 201, 183–186 (1986).

  35. 35.

    Ng, R. et al. Inhibition of microRNA-24 expression in liver prevents hepatic lipid accumulation and hyperlipidemia. Hepatology 60, 554–564 (2014).

  36. 36.

    Tao, R., Xiong, X., DePinho, R. A., Deng, C.-X. & Dong, X. C. Hepatic SREBP-2 and cholesterol biosynthesis are regulated by FoxO3 and Sirt6. J. Lipid Res. 54, 2745–2753 (2013).

  37. 37.

    Beaulieu, J. M. et al. An Akt/β-arrestin 2/PP2A signaling complex mediates dopaminergic neurotransmission and behavior. Cell 122, 261–273 (2005).

  38. 38.

    Lefkowitz, R. J. & Shenoy, S. K. Transduction of receptor signals by ß-arrestins. Science 308, 512–517 (2005).

  39. 39.

    Yang, F. et al. Phospho-selective mechanisms of arrestin conformations and functions revealed by unnatural amino acid incorporation and 19F-NMR. Nat. Commun. 6, 8202 (2015).

  40. 40.

    Pedram, A. et al. Estrogen reduces lipid content in the liver exclusively from membrane receptor signaling. Sci. Signal. 6, ra36–ra36 (2013).

  41. 41.

    Oka, R. et al. Relationships between alanine aminotransferase (ALT), visceral adipose tissue (AT) and metabolic risk factors in a middle-aged Japanese population. J. Atheroscler. Thromb. 21, 582–592 (2014).

  42. 42.

    Choi, Y. et al. Menopausal stages and serum lipid and lipoprotein abnormalities in middle-aged women. Maturitas 80, 399–405 (2015).

  43. 43.

    Grundy, S. M. HMG-CoA reductase inhibitors for treatment of hypercholesterolemia. N. Engl. J. Med. 319, 24–33 (1988).

  44. 44.

    Fak, A. S. et al. Effects of simvastatin only or in combination with continuous combined hormone replacement therapy on serum lipid levels in hypercholesterolaemic post-menopausal women. Eur. Heart J. 21, 190–197 (2000).

  45. 45.

    Lemay, A., Dodin, S., Turcot, L., Déchêne, F. & Forest, J. C. Estrogen/progesterone replacement versus pravastatin and their sequential association in hypercholesterolemic postmenopausal women. Maturitas 40, 247–257 (2001).

  46. 46.

    Serruys, P. W. et al. A randomized placebo-controlled trial of fluvastatin for prevention of restenosis after successful coronary balloon angioplasty: final results of the fluvastatin angiographic restenosis (FLARE) trial. Eur. Heart J. 20, 58–69 (1999).

  47. 47.

    Bradford, R. H. et al. Efficacy and tolerability of lovastatin in 3390 women with moderate hypercholesterolemia. Ann. Intern. Med. 118, 850–855 (1993).

  48. 48.

    Thurston, R. C. et al. Gains in body fat and vasomotor symptom reporting over the menopausal transition: the Study of Women’s Health Across the Nation. Am. J. Epidemiol. 170, 766–774 (2009).

  49. 49.

    Haring, B. et al. Cardiovascular disease and cognitive decline in postmenopausal women: results from the Women’s Health Initiative Memory Study. J. Am. Heart Assoc. 2, e000369 (2013).

  50. 50.

    van den Berghe, G. in Journal of Inherited Metabolic Disease (eds R. A. Harkness, R. J. Pollitt, & G. M. Addison) 407–420 (Springer Netherlands, 1991).

  51. 51.

    Nakagawa, Y. & Shimano, H. CREBH regulates systemic glucose and lipid metabolism. Int. J. Mol. Sci. 19, 1396 (2018).

  52. 52.

    Mota, M., Banini, B. A., Cazanave, S. C. & Sanyal, A. J. Molecular mechanisms of lipotoxicity and glucotoxicity in nonalcoholic fatty liver disease. Metabolism 65, 1049–1061 (2016).

  53. 53.

    Qi, X. et al. Follicle-stimulating hormone enhances hepatic gluconeogenesis by GRK2-mediated AMPK hyperphosphorylation at Ser485 in mice. Diabetologia 61, 1180–1192 (2018).

  54. 54.

    Li, Y. et al. A global perspective on FOXO1 in lipid metabolism and lipid-related diseases. Prog. Lipid Res. 66, 42–49 (2017).

  55. 55.

    Matsumoto, M., Han, S., Kitamura, T. & Accili, D. Dual role of transcription factor FoxO1 in controlling hepatic insulin sensitivity and lipid metabolism. J. Clin. Invest. 116, 2464–2472 (2006).

  56. 56.

    Lee, S. & Dong, H. H. FoxO integration of insulin signaling with glucose and lipid metabolism. J. Endocrinol. 233, R67–R79 (2017).

  57. 57.

    Smith, B. K. & Steinberg, G. R. AMP-activated protein kinase, fatty acid metabolism, and insulin sensitivity. Curr. Opin. Clin. Nutr. Metab. Care 20, 248–253 (2017).

  58. 58.

    Gopoju, R., Panangipalli, S. & Kotamraju, S. Metformin treatment prevents SREBP2-mediated cholesterol uptake and improves lipid homeostasis during oxidative stress-induced atherosclerosis. Free Radic. Biol. Med. 118, 85–97 (2018).

  59. 59.

    Zhang, B. B., Zhou, G. & Li, C. AMPK: an emerging drug target for diabetes and the metabolic syndrome. Cell. Metab. 9, 407–416 (2009).

  60. 60.

    Oppenheim, D. S., Greenspan, S. L., Zervas, N. T., Schoenfeld, D. A. & Anne Klibanski, M. Elevated serum lipids in hypogonadal men with and without hyperprolactinemia. Ann. Intern. Med. 111, 288–92 (1989).

  61. 61.

    Derby, C. A. et al. Lipid changes during the menopause transition in relation to age and weight: the Study of Women’s Health Across the Nation. Am. J. Epidemiol. 169, 1352–1361 (2009).

  62. 62.

    Nagaraj, N. et al. Complement proteins and arterial calcification in middle aged women: cross-sectional effect of cardiovascular fat. The SWAN Cardiovascular Fat Ancillary Study. Atherosclerosis 243, 533–539 (2015).

  63. 63.

    Luepker, R. V., Evans, A., McKeigue, P. & Reddy, K. S. in Cardiovascular Survey Methods (World Health Organization, Geneva, 2004).

  64. 64.

    Yang, W. et al. Prevalence of diabetes among men and women in China. N. Engl. J. Med. 362, 1090–1101 (2010).

  65. 65.

    Guidelines for the management of dyslipidaemia in chinese adults. Chin. Circ. J. 31, 937–953 (2016).

  66. 66.

    Expert Panel on, D. Evaluation, and Treatment of High Blood Cholesterol in Adults. Executive summary of the 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). JAMA 285, 2486–2497 (2001).

  67. 67.

    Fafalios, A. et al. A hepatocyte growth factor receptor (Met)−insulin receptor hybrid governs hepatic glucose metabolism. Nat. Med. 17, 1577 (2011).

  68. 68.

    Li, Y. et al. A novel role for CRTC2 in hepatic cholesterol synthesis through SREBP-2. Hepatology 66, 481–497 (2017).

  69. 69.

    Zhang, X. et al. Thyroid-stimulating hormone decreases HMG-CoA reductase phosphorylation via AMP-activated protein kinase in the liver. J. Lipid Res. 56, 963–971 (2015).

  70. 70.

    Aryal, B. et al. Absence of ANGPTL4 in adipose tissue improves glucose tolerance and attenuates atherogenesis. JCI Insight 3, e97918 (2018).

  71. 71.

    Prensner, J. R. et al. The long noncoding RNA SChLAP1 promotes aggressive prostate cancer and antagonizes the SWI/SNF complex. Nat. Genet. 45, 1392 (2013).

  72. 72.

    Abe, E. et al. TSH is a negative regulator of skeletal remodeling. Cell 115, 151–162 (2003).

  73. 73.

    Wang, Y. et al. Tomato nuclear proteome reveals the involvement of specific E2 ubiquitin-conjugating enzymes in fruit ripening. Genome Biol. 15, 548 (2014).

  74. 74.

    Roberts, T. C. et al. Quantification of nascent transcription by bromouridine immunocapture nuclear run-on RT-qPCR. Nat. Protoc. 10, 1198 (2015).

  75. 75.

    Dentin, R. et al. Insulin modulates gluconeogenesis by inhibition of the coactivator TORC2. Nature 449, 366 (2007).

  76. 76.

    Honda, A. et al. Differences in hepatic levels of intermediates in bile acid biosynthesis between Cyp27−/− mice and CTX. J. Lipid Res. 42, 291–300 (2001).

  77. 77.

    Honda, A. et al. Highly sensitive assay of HMG-CoA reductase activity by LC-ESI-MS/MS. J. Lipid Res. 48, 1212–1220 (2007).

  78. 78.

    Li, T. et al. Overexpression of cholesterol 7α-hydroxylase promotes hepatic bile acid synthesis and secretion and maintains cholesterol homeostasis. Hepatology 53, 996–1006 (2011).

  79. 79.

    Wang, H. M. et al. A stress response pathway in mice upregulates somatostatin level and transcription in pancreatic delta cells through Gs and β-arrestin 1. Diabetologia 57, 1899–1910 (2014).

  80. 80.

    Song, Y. et al. Thyroid-stimulating hormone regulates hepatic bile acid homeostasis via SREBP-2/HNF-4α/CYP7A1 axis. J. Hepatol. 62, 1171–1179 (2015).

Download references

Acknowledgements

We thank Prof. Yingli Lu (Shanghai JiaoTong University School of Medicine, China) for the assistance in the measurement of de novo cholesterol biosynthesis. Great thanks to Prof. Jinpeng Sun and Prof. Xiao Yu (Shandong University School of Medicine, China) for their technical assistance referring to G proteins and β-arrestins in this project. We thank Shengfeng Wang (Department of Epidemiology and Biostatistics, Peking University Health Science Center, China) for providing professional instructions in statistical analyses. We are grateful to Prof. Yongsheng Chang (Chinese Academy of Medical Sciences and Peking Union Medical College) for his suggestions in writing the manuscript. And great thanks to Prof. Shucun Qin and his team (Taishan Medical University, China). This work was supported by grants from the National Natural Science Foundation (81870607, 81670796, 81430020, 81230018, 81300644 and 31640020) and National Key R&D Program (SQ2017YFSF090203) of China.

Author information

Affiliations

  1. Department of Endocrinology, Shandong Provincial Hospital Affiliated to Shandong University, 250021, Jinan, Shandong, China

    • Yanjing Guo
    • , Meng Zhao
    • , Shizhan Ma
    • , Lu Liu
    • , Yujie Li
    • , Chunxiao Yu
    • , Xiaoyi Qi
    • , Qian Wang
    • , Haiqing Zhang
    • , Qingbo Guan
    • , Lifang Zhao
    • , Shimeng Xuan
    • , Huili Yan
    • , Qihang Li
    • , Yongfeng Song
    •  & Jiajun Zhao
  2. Shandong Key Laboratory of Endocrinology and Lipid Metabolism, 250021, Jinan, Shandong, China

    • Yanjing Guo
    • , Meng Zhao
    • , Shizhan Ma
    • , Zhao He
    • , Lu Liu
    • , Yujie Li
    • , Chunxiao Yu
    • , Xiaoyi Qi
    • , Qian Wang
    • , Haiqing Zhang
    • , Qingbo Guan
    • , Lifang Zhao
    • , Shimeng Xuan
    • , Huili Yan
    • , Qihang Li
    • , Yongfeng Song
    •  & Jiajun Zhao
  3. Institute of Endocrinology and metabolism, Shandong Academy of Clinical Medicine, 250021, Jinan, Shandong, China

    • Yanjing Guo
    • , Meng Zhao
    • , Shizhan Ma
    • , Zhao He
    • , Lu Liu
    • , Yujie Li
    • , Chunxiao Yu
    • , Xiaoyi Qi
    • , Qian Wang
    • , Haiqing Zhang
    • , Qingbo Guan
    • , Lifang Zhao
    • , Shimeng Xuan
    • , Huili Yan
    • , Qihang Li
    • , Yongfeng Song
    • , Ling Gao
    •  & Jiajun Zhao
  4. Scientific Center, Shandong Provincial Hospital Affiliated to Shandong University, 250021, Jinan, Shandong, China

    • Tao Bo
    • , Wenbin Chen
    • , Xu Hou
    • , Yanliang Lin
    •  & Ling Gao
  5. Department of Biostatistics, School of Public Health, Shandong University, 250012, Jinan, Shandong, China

    • Zhongshang Yuan
  6. Department of Hepatobiliary Surgery, Shandong Provincial Hospital Affiliated to Shandong University, 250021, Jinan, Shandong, China

    • Jun Liu
    •  & Zhenhai Zhang
  7. Department of Gastroenterology, Shandong Provincial Hospital Affiliated to Shandong University, 250021, Jinan, Shandong, China

    • Qiang Zhu
  8. Department of Pathology, Shandong Provincial Hospital Affiliated to Shandong University, 250021, Jinan, Shandong, China

    • Qiangxiu Wang
    •  & Xiaoyan Lin
  9. Department of Gynecology, Shandong Provincial Hospital Affiliated to Shandong University, 250021, Jinan, Shandong, China

    • Zhongli Yang
    •  & Min Cui
  10. Department of Physiology and Neurobiology, and Institute for Systems Genomics, University of Connecticut, Storrs, CT, 06269, USA

    • Li Wang
    •  & Yongfeng Song

Authors

  1. Search for Yanjing Guo in:

  2. Search for Meng Zhao in:

  3. Search for Tao Bo in:

  4. Search for Shizhan Ma in:

  5. Search for Zhongshang Yuan in:

  6. Search for Wenbin Chen in:

  7. Search for Zhao He in:

  8. Search for Xu Hou in:

  9. Search for Jun Liu in:

  10. Search for Zhenhai Zhang in:

  11. Search for Qiang Zhu in:

  12. Search for Qiangxiu Wang in:

  13. Search for Xiaoyan Lin in:

  14. Search for Zhongli Yang in:

  15. Search for Min Cui in:

  16. Search for Lu Liu in:

  17. Search for Yujie Li in:

  18. Search for Chunxiao Yu in:

  19. Search for Xiaoyi Qi in:

  20. Search for Qian Wang in:

  21. Search for Haiqing Zhang in:

  22. Search for Qingbo Guan in:

  23. Search for Lifang Zhao in:

  24. Search for Shimeng Xuan in:

  25. Search for Huili Yan in:

  26. Search for Yanliang Lin in:

  27. Search for Li Wang in:

  28. Search for Qihang Li in:

  29. Search for Yongfeng Song in:

  30. Search for Ling Gao in:

  31. Search for Jiajun Zhao in:

Contributions

Y.J.G. and Y.F.S. performed most of the experiments, analyzed and interpreted data and wrote the manuscript. T.B. helped to construct the Fshr-/- mouse model and design some primers. H.Q.Z., M.Z., Z.S.Y., and L.L. conducted and analyzed the epidemiological data. M.S.Z. and W.B.C. performed the bioinformatic analysis. J.L. and Z.H.Z. provided the human liver samples. Z.L.Y. and M.C. provided the human ovarian samples. Q.X.W. and X.Y.L. helped to analyze the pathology results. M.X.S. and Q.H.L. helped to maintain the animal models. H.L.Y., and L.F.Z., performed some of the in vitro experiments. Q.W., C.X.Y., X.Y.Q., Y.L.L., and Y.J.L. helped with some of the in vivo experiments. Z.H., X.H., and L.W. helped to write the manuscript. L.G. and J.J.Z supervised the project and wrote the manuscript.

Competing interests

The authors declare no competing interests.

Corresponding authors

Correspondence to Yongfeng Song or Ling Gao or Jiajun Zhao.

Electronic supplementary material

About this article

Publication history

Received

Accepted

Published

Issue Date

DOI

https://doi.org/10.1038/s41422-018-0123-6