Salt-sensitive hypertension in circadian clock–deficient Cry-null mice involves dysregulated adrenal Hsd3b6

Abstract

Malfunction of the circadian clock has been linked to the pathogenesis of a variety of diseases. We show that mice lacking the core clock components Cryptochrome-1 (Cry1) and Cryptochrome-2 (Cry2) (Cry-null mice) show salt-sensitive hypertension due to abnormally high synthesis of the mineralocorticoid aldosterone by the adrenal gland. An extensive search for the underlying cause led us to identify type VI 3β-hydroxyl-steroid dehydrogenase (Hsd3b6) as a new hypertension risk factor in mice. Hsd3b6 is expressed exclusively in aldosterone-producing cells and is under transcriptional control of the circadian clock. In Cry-null mice, Hsd3b6 messenger RNA and protein levels are constitutively high, leading to a marked increase in 3β-hydroxysteroid dehydrogenase-isomerase (3β-HSD) enzymatic activity and, as a consequence, enhanced aldosterone production. These data place Hsd3b6 in a pivotal position through which circadian clock malfunction is coupled to the development of hypertension. Translation of these findings to humans will require clinical examination of human HSD3B1 gene, which we found to be functionally similar to mouse Hsd3b6.

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Figure 1: Overproduction of aldosterone by Cry-null adrenal glands.
Figure 2: Chronically high expression of Hsd3b6 in aldosterone-producing zona glomerulosa (ZG) cells of Cry-null adrenal glands.
Figure 3: Coexpression of Hsd3b6 protein and Cyp11b2 mRNA in adrenal zona glomerulosa cells.
Figure 4: Elevated 3β-HSD activity is responsible for aldosterone overproduction in Cry-null mice.
Figure 5: Cry-null mice show salt-sensitive hypertension.
Figure 6: Identification of a zona glomerulosa–specific HSD3B isoform in the human adrenal gland.

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Acknowledgements

We thank T. Ono, A. Hirasawa, T. Koshimizu, K. Terasawa, H. Nishinaga, M. Sato, Y. Yamaguchi, M. Matsuo, J.M. Fustin, K. Toida, H. Sei and K. Ishimura for technical support and valuable discussion. We also thank T. Michel for critical reading of the manuscript. This work was supported in part by the Specially Promoted Research (to H.O.) and Grant-in-Aid for Young Scientists (to M.D.) from the Ministry of Education, Culture, Sports, Science and Technology of Japan and grants from Nakatomi Foundation, SRF (to H.O.), Senri Life Science Foundation, Takeda Science Foundation (to M.D.) and the Netherlands Organization of Scientific research, ZonMW Vici 918.36.619 (to G.T.J.v.d.H.). Trilostane was a generous gift from Mochida Pharmaceutical. Eplerenone was a generous gift from Pfizer.

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M.D. and H.O. designed the research; A.K., O.O., T.T. and G.T.J.v.d.H. supplied the experimental materials; M.D., Y.T., R.K., F.Y., H.Y., S.H., K.T. and H.O. acquired the data; M.D., N.E., Y.O., G.T., K.T. and H.O. analyzed the data; and M.D., G.T.J.v.d.H. and H.O. drafted the manuscript.

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Correspondence to Hitoshi Okamura.

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Doi, M., Takahashi, Y., Komatsu, R. et al. Salt-sensitive hypertension in circadian clock–deficient Cry-null mice involves dysregulated adrenal Hsd3b6. Nat Med 16, 67–74 (2010). https://doi.org/10.1038/nm.2061

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