Institute of Molecular and Cell Biology, Proteos, Singapore.
Institute of Bioengineering and Nanotechnology, Nanos, Singapore.
Stem Cells Transl Med. 2017 Mar;6(3):851-863. doi: 10.5966/sctm.2016-0129. Epub 2016 Oct 7.
Atherosclerosis underlies many cardiovascular and cerebrovascular diseases. Nutraceuticals are emerging as a therapeutic moiety for restoring vascular health. Unlike small-molecule drugs, the complexity of ingredients in nutraceuticals often confounds evaluation of their efficacy in preclinical evaluation. It is recognized that the liver is a vital organ in processing complex compounds into bioactive metabolites. In this work, we developed a coculture system of human pluripotent stem cell-derived endothelial cells (hPSC-ECs) and human pluripotent stem cell-derived hepatocytes (hPSC-HEPs) for predicting vascular-protective effects of nutraceuticals. To validate our model, two compounds (quercetin and genistein), known to have anti-inflammatory effects on vasculatures, were selected. We found that both quercetin and genistein were ineffective at suppressing inflammatory activation by interleukin-1β owing to limited metabolic activity of hPSC-ECs. Conversely, hPSC-HEPs demonstrated metabolic capacity to break down both nutraceuticals into primary and secondary metabolites. When hPSC-HEPs were cocultured with hPSC-ECs to permit paracrine interactions, the continuous turnover of metabolites mitigated interleukin-1β stimulation on hPSC-ECs. We observed significant reductions in inflammatory gene expressions, nuclear translocation of nuclear factor κB, and interleukin-8 production. Thus, integration of hPSC-HEPs could accurately reproduce systemic effects involved in drug metabolism in vivo to unravel beneficial constituents in nutraceuticals. This physiologically relevant endothelial-hepatic platform would be a great resource in predicting the efficacy of complex nutraceuticals and mechanistic interrogation of vascular-targeting candidate compounds. Stem Cells Translational Medicine 2017;6:851-863.
动脉粥样硬化是许多心血管和脑血管疾病的基础。营养保健品作为一种恢复血管健康的治疗药物正在兴起。与小分子药物不同,营养保健品成分的复杂性常常使人们难以评估其在临床前评估中的疗效。人们认识到,肝脏是将复杂化合物转化为生物活性代谢物的重要器官。在这项工作中,我们开发了一种人多能干细胞衍生的内皮细胞(hPSC-ECs)和人多能干细胞衍生的肝细胞(hPSC-HEPs)共培养系统,用于预测营养保健品的血管保护作用。为了验证我们的模型,选择了两种已知对血管具有抗炎作用的化合物(槲皮素和染料木黄酮)。我们发现,由于 hPSC-ECs 的代谢活性有限,槲皮素和染料木黄酮都不能抑制白细胞介素-1β引起的炎症激活。相反,hPSC-HEPs 表现出将两种营养保健品分解为初级和次级代谢物的代谢能力。当 hPSC-HEPs 与 hPSC-ECs 共培养以允许旁分泌相互作用时,代谢物的连续转化减轻了白细胞介素-1β对 hPSC-ECs 的刺激。我们观察到炎症基因表达、核因子κB 的核转位和白细胞介素-8 产生的显著减少。因此,整合 hPSC-HEPs 可以准确再现体内药物代谢涉及的全身效应,从而揭示营养保健品中的有益成分。这种具有生理相关性的内皮-肝平台将成为预测复杂营养保健品疗效和血管靶向候选化合物的机制探究的重要资源。《干细胞转化医学》2017 年;6:851-863。