a locked nucleic acid antisense oligonucleotide (lna) silences pcsk9 and enhances ldlr expression in vitro and in vivo一个锁定的核酸反义寡核苷酸(放大器)沉默pcsk9和增强ldlr表达在体外和体内.pdfVIP

A Locked Nucleic Acid Antisense Oligonucleotide (LNA) Silences PCSK9 and Enhances LDLR Expression In Vitro and In Vivo 1. 2. 1 2 2 Nidhi Gupta , Niels Fisker , Marie-Claude Asselin , Marie Lindholm , Christoph Rosenbohm , Henrik 2 ´ 2 1 2 Ørum , Joacim Elmen , Nabil G. Seidah * , Ellen Marie Straarup 1 Laboratory of Biochemical Neuroendocrinology, Clinical Research Institute of Montreal, Montreal, Quebec, Canada, 2 Santaris Pharma A/S, Hørsholm, Denmark Abstract Background: The proprotein convertase subtilisin/kexin type 9 (PCSK9) is an important factor in the etiology of familial hypercholesterolemia (FH) and is also an attractive therapeutic target to reduce low density lipoprotein (LDL) cholesterol. PCSK9 accelerates the degradation of hepatic low density lipoprotein receptor (LDLR) and low levels of hepatic PCSK9 activity are associated with reduced levels of circulating LDL-cholesterol. Methodology/Principal Findings: The present study presents the first evidence for the efficacy of a locked nucleic acid (LNA) antisense oligonucleotide (LNA ASO) that targets both human and mouse PCSK9. We employed human hepatocytes derived cell lines HepG2 and HuH7 and a pancreatic mouse b-TC3 cell line known to express high endogenous levels of PCSK9. LNA ASO efficiently reduced the mRNA and protein levels of PCSK9 with a concomitant increase in LDLR protein levels after transfection in these cells. In vivo efficacy of LNA ASO was further investigated in mice by tail vein intravenous administration of LNA ASO in saline solution. The level of PCSK9 mRNA was reduced by ,60%, an effect lasting more than