*Article* **Role of Cryptochrome-1 and Cryptochrome-2 in Aldosterone-Producing Adenomas and Adrenocortical Cells**

**Martina Tetti 1, Isabella Castellano 2, Francesca Veneziano 2, Corrado Magnino 1, Franco Veglio 1, Paolo Mulatero <sup>1</sup> and Silvia Monticone 1,\***


Received: 7 April 2018; Accepted: 31 May 2018; Published: 5 June 2018

**Abstract:** Mice lacking the core-clock components, cryptochrome-1 (CRY1) and cryptochrome-2 (CRY2) display a phenotype of hyperaldosteronism, due to the upregulation of type VI 3β-hydroxyl-steroid dehydrogenase (*Hsd3b6*), the murine counterpart to the human type I 3β-hydroxyl-steroid dehydrogenase (*HSD3B1*) gene. In the present study, we evaluated the role of *CRY1* and *CRY2* genes, and their potential interplay with *HSD3B* isoforms in adrenal pathophysiology in man. Forty-six sporadic aldosterone-producing adenomas (APAs) and 20 paired adrenal samples were included, with the human adrenocortical cells HAC15 used as the in vitro model. In our cohort of sporadic APAs, *CRY1* expression was 1.7-fold [0.75–2.26] higher (*p* = 0.016), while *CRY2* showed a 20% lower expression [0.80, 0.52–1.08] (*p* = 0.04) in APAs when compared with the corresponding adjacent adrenal cortex. Type II 3β-hydroxyl-steroid dehydrogenase (*HSD3B2*) was 317-fold [200–573] more expressed than *HSD3B1*, and is the main *HSD3B* isoform in APAs. Both dehydrogenases were more expressed in APAs when compared with the adjacent cortex (5.7-fold and 3.5-fold, respectively, *p* < 0.001 and *p* = 0.001) and *HSD3B1* was significantly more expressed in APAs composed mainly of zona glomerulosa-like cells. Treatment with angiotensin II (AngII) resulted in a significant upregulation of *CRY1* (1.7 ± 0.25-fold, *p* < 0.001) at 6 h, and downregulation of *CRY2* at 12 h (0.6 ± 0.1-fold, *p* < 0.001), through activation of the AngII type 1 receptor. Independent silencing of *CRY1* and *CRY2* genes in HAC15 cells resulted in a mild upregulation of *HSD3B2* without affecting *HSD3B1* expression. In conclusion, our results support the hypothesis that *CRY1* and *CRY2*, being AngII-regulated genes, and showing a differential expression in APAs when compared with the adjacent adrenal cortex, might be involved in adrenal cell function, and in the regulation of aldosterone production.

**Keywords:** aldosterone-producing adenoma; *CRY1*; *CRY2*; *HSD3B1*; *HSD3B2*
