The Response Of An In Vitro Cynomolgus Macaque System To Statin Treatments Mirrors The Human In Vitro System And Clinical Outcomes

Introduction: There is an urgent unmet need to improve the predictive value of animal and in vitro systems for drug development. The purpose of this study was to develop an in vitro vascular system that reflects the in vivo biology of the cynomolgus macaque (cyno). Methods: We co-cultured endothelial (EC) and smooth muscle cells (SMC) from cyno arteries and exposed them to physiologically relevant hemodynamics. Cyno velocity profiles were obtained from arterial regions prone to atherosclerosis (atheroprone) or healthy regions. Velocity waveforms were applied to the co-culture along with oxidized LDL from cyno or humans, and TNFα, which are pathogenic in humans and mimic advanced inflammatory conditions (AIC). Using RNA sequencing, we quantified the response of the system to 4 different statin treatments (atorvastatin, simvastatin, cerivastatin, rosuvastatin) under AIC. Using computational analyses of differentially expressed genes (DEGs), we characterized the response of our system by identifying functional biological themes that were associated with each statin treatment. Results: We found that AIC in the cyno system dramatically increased inflammatory DEGs and decreased vascular health DEGs. Cerivastatin elicited the most DEGs in both the cyno and human vascular system while the response to rosuvastatin did not reach statistical significance for any genes. Statin treatment decreased inflammatory cytokine gene expression, but not adhesion molecule expression under AIC. Except for rosuvastatin, all statins tested significantly increased the expression of genes that promote vascular health, while suppressing the expression of inflammatory genes. Biological themes were regulated by statin treatment in both the human and cyno vascular systems including: cholesterol biosynthesis, thrombosis, ethanol degradation, cell cycle regulation and notch signaling. Finally, we discovered functional themes related to glycogen metabolism, which may be relevant as a potential mechanism driving the risk of hyperglycemia with statin treatment. Conclusions: The cyno vascular system described here mirrors many of the well-known phenotypes of the human vascular system, and should provide a valuable predictive tool for in vivo studies.