Abstract 402: miRNA Targeting of Oxysterol Binding Protein-like 6 (OSBPL6) Regulates Cholesterol Trafficking and Efflux

Abstract

Cholesterol homeostasis is fundamental to human health, and is thus, tightly regulated. MicroRNAs exert potent effects on biological pathways, including cholesterol metabolism, by repressing genes with related functions. We reasoned that this mode of pathway regulation could be exploited to identify novel genes involved in cholesterol homeostasis. Here, we identify oxysterol binding protein-like 6 ( OSBPL6 ) as a novel target of 2 miRNA hubs regulating cholesterol homeostasis: miR-33 and miR-27b. Characterization of OSBPL6 revealed that it is transcriptionally regulated in macrophages and hepatocytes by LXR and in response to cholesterol loading, and in mice and non-human primates by western diet feeding. OSBPL6 encodes the OSBPL-related protein 6 (ORP6), which contains a pleckstrin homology membrane-targeting domain and a FFAT domain predicted to interact with VAP proteins in the ER. Indeed, subcellular localization studies showed that ORP6 is associated with the plasma membrane, early endosomes, lysosomes and ER, suggesting a putative role for ORP6 in cholesterol trafficking between these compartments. Consistent with this, siRNA-mediated knockdown of OSBPL6 results in aberrant clustering of endosomes and promotes the accumulation of free cholesterol in these structures. As a consequence, esterification of cholesterol at the ER is reduced, triggering activation of SREBP2. Conversely, ORP6 overexpression enhances cholesterol trafficking and efflux to apoA1 in macrophages and hepatocytes. Moreover, we show that hepatic expression of OSBPL6 is positively correlated with plasma levels of HDL cholesterol in a cohort of 200 healthy individuals, whereas its expression is reduced in human atherosclerotic plaques. Collectively, these studies identify ORP6 as a novel regulator of cholesterol trafficking that is part of the miR-33 and miR-27b target gene networks that contribute to the maintenance of cholesterol homeostasis.