27-Hydroxylation of oncosterone by CYP27A1 switches its activity from pro-tumor to anti-tumor

Oncosterone (6-oxo-cholestane-3 beta,5 alpha-diol; OCDO) is an oncometabolite and a tumor promoter on estrogen receptor alpha-positive breast cancer (ER(+) BC) and triple-negative breast cancers (TN BC). OCDO is an oxysterol formed in three steps from cholesterol: 1) oxygen addition at the double bond to give alpha-or beta-isomers of 5,6-epoxycholestanols (5,6-EC), 2) hydrolyses of the epoxide ring of 5,6-ECs to give cholestane-3 beta,5 alpha,6 beta-triol (CT), and 3) oxidation of the C6 hydroxyl of CT to give OCDO. On other hand, cholesterol can be hydroxylated CYP27A1 at the ultimate methyl carbon of its side chain to give 27-hydroxycholesterol ((25R)-Cholest-5-ene-3beta,26-diol, 27HC), which is a tumor promoter for ER(+) BC. It is currently unknown whether OCDO and its precursors can be hydroxylated position C27 by CYP27A1, as is the impact of such modification on the proliferation of ER(+) and TN BC cells. We investigated, herein, whether 27H-5,6-ECs ((25R)-5,6-epoxycholestan-3 beta,26-diol), 27H-CT ((25R)-cholestane-3 beta,5 alpha,6 beta,26-tetrol) and 27H-OCDO ((25R)-cholestane-6-oxo-3 beta,5 alpha,26-triol) exist as metabolites and can be produced by cells expressing CYP27A1. We report, for the first time, that these compounds exist as metabolites in humans. We give pharmacological and genetic evidence that CYP27A1 is responsible for their production. Importantly, we found that 27-hydroxy-OCDO (27H-OCDO) inhibits BC cell proliferation and blocks OCDO and HC-induced proliferation in BC cells, showing that this metabolic conversion commutes the proliferative properties of OCDO into antiproliferative ones. These data suggest an unprecedented role CYP27A1 in the control of breast carcinogenesis inhibiting the tumor promoter activities of oncosterone and 27-HC.