Abstract 168: Effects of the CETP Inhibitor Anacetrapib on HDL3-to-HDL2 Transfer: Comparison of in Vitro and in Vivo Methodologies

Cholesteryl ester transfer protein (CETP) is a target for the treatment of dyslipidemia and coronary artery disease. In addition to the well-known effect of CETP to transfer CE from HDL to LDL and to VLDL, in vitro , CETP has been reported to transfer CE between small and large HDL particles (HDL2 and HDL3, respectively). We sought to understand how the CETP inhibitor anacetrapib (ANA) affects HDL3-to-HDL2 transfer under both in vitro and in vivo conditions. In vitro , ANA dose-dependently inhibited transfer of 3 H-CE from total HDL to LDL (IC50 30nM), and from isolated HDL3 to HDL2 particles (IC50 200nM). In human CETP transgenic mice, animals treated with a single dose of ANA (100mg/kg) displayed 80% maximal reduction in plasma CETP activity and a 22% increase in total HDL cholesterol. In animals treated with either vehicle or ANA, 3 H-CE-labeled HDL3 was injected intravenously and 3 H-tracer was monitored in lipoprotein fractions following injection. Animals treated with ANA showed an increase in the amount 3 H-tracer present in HDL2 compared to vehicle over time (20-70% increase across 6 hrs post 3 H-CE-HDL3 injection, P<0.05 vs vehicle). The HDL2 CE pool was also increased with ANA treatment, and 3 H-cholesterol flux into HDL2 was increased with ANA treatment when adjusted to the change in pool size (at 2 and 4 hrs post 3H-CE-HDL3 injection). No change in HDL2 3 H-tracer was seen in C57BL6 mice (lacking CETP) treated with ANA. These results indicate that in contrast to in vitro findings, ANA increases flux of CE into HDL2 in vivo , a process which likely involves multiple pathways. Therefore, the in vitro phenomena of 1) HDL3-to-HDL2 transfer by CETP and 2) inhibition of this transfer by CETP inhibitors are not recapitulated in vivo . It is clear that in vivo approaches are necessary to understand the relevance of HDL3-to-HDL2 transfer in vivo , and to accurately study the effects of CETP inhibition on lipoprotein metabolism.