Department of Cardiovascular Surgery of the First Affiliated Hospital & Institute for Cardiovascular Science, Collaborative Innovation Center of Hematology, State Key Laboratory of Radiation Medicine and Protection, Medical College, Soochow University, Suzhou, 215000, China.
Institute of Neuroscience, Soochow University, Suzhou, 215123, China.
Acta Pharmacol Sin. 2022 Jan;43(1):240-250. doi: 10.1038/s41401-021-00621-8. Epub 2021 Mar 8.
Cardiovascular safety assessment is vital for drug development, yet human cardiovascular cell models are lacking. In vitro mass-generated human pluripotent stem cell (hPSC)-derived cardiovascular cells are a suitable cell model for preclinical cardiovascular safety evaluations. In this study, we established a preclinical toxicology model using same-origin hPSC-differentiated cardiomyocytes (hPSC-CMs) and endothelial cells (hPSC-ECs). For validation of this cell model, alirocumab, a human antibody against proprotein convertase subtilisin kexin type 9 (PCSK9), was selected as an emerging safe lipid-lowering drug; atorvastatin, a common statin (the most effective type of lipid-lowering drug), was used as a drug with reported side effects at high concentrations, while doxorubicin was chosen as a positive cardiotoxic drug. The cytotoxicity of these drugs was assessed using CCK8, ATP, and lactate dehydrogenase release assays at 24, 48, and 72 h. The influences of these drugs on cardiomyocyte electrophysiology were detected using the patch-clamp technique, while their effects on endothelial function were determined by tube formation and Dil-acetylated low-density lipoprotein (Dil-Ac-LDL) uptake assays. We showed that alirocumab did not affect the cell viability or cardiomyocyte electrophysiology in agreement with the clinical results. Atorvastatin (5-50 μM) dose-dependently decreased cardiovascular cell viability over time, and at a high concentration (50 μM, ~100 times the normal peak serum concentration in clinic), it affected the action potentials of hPSC-CMs and damaged tube formation and Dil-Ac-LDL uptake of hPSC-ECs. The results demonstrate that the established same-origin hPSC-derived cardiovascular cell model can be used to evaluate lipid-lowering drug safety in cardiovascular cells and allow highly accurate preclinical assessment of potential drugs.
心血管安全性评估对于药物开发至关重要，但目前缺乏人类心血管细胞模型。体外大量生成的人多能干细胞（hPSC）衍生的心血管细胞是进行临床前心血管安全性评估的合适细胞模型。在本研究中，我们使用同源 hPSC 分化的心肌细胞（hPSC-CMs）和内皮细胞（hPSC-ECs）建立了临床前毒理学模型。为了验证该细胞模型，我们选择了一种新型降脂药——针对前蛋白转化酶枯草溶菌素 9（PCSK9）的人源单克隆抗体阿里西尤单抗；阿托伐他汀是一种常见的他汀类药物（最有效的降脂药之一），我们用它来验证高浓度下药物的已知副作用；阿霉素则被选为阳性心脏毒性药物。我们使用 CCK8、ATP 和乳酸脱氢酶释放测定法在 24、48 和 72 小时评估这些药物的细胞毒性。使用膜片钳技术检测这些药物对心肌细胞电生理的影响，通过管形成和 Dil-乙酰化低密度脂蛋白（Dil-Ac-LDL）摄取测定法检测其对内皮功能的影响。结果表明，与临床结果一致，阿里西尤单抗不影响细胞活力或心肌细胞电生理。阿托伐他汀（5-50 μM）随时间呈剂量依赖性降低心血管细胞活力，在高浓度（50 μM，约为临床正常峰值血清浓度的 100 倍）时，它会影响 hPSC-CMs 的动作电位，并破坏 hPSC-ECs 的管形成和 Dil-Ac-LDL 摄取。结果表明，建立的同源 hPSC 衍生心血管细胞模型可用于评估降脂药在心血管细胞中的安全性，并允许对潜在药物进行高度准确的临床前评估。
