Mina Sara G, Alaybeyoglu Begum, Murphy William L, Thomson James A, Stokes Cynthia L, Cirit Murat
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, United States.
Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, United States.
Front Big Data. 2019 Jun 26;2:23. doi: 10.3389/fdata.2019.00023. eCollection 2019.
Early assessment of adverse drug effects in humans is critical to avoid long-lasting harm. However, current approaches for early detection of adverse effects still lack predictive and organ-specific biomarkers to evaluate undesired responses in humans. Microphysiological systems (MPSs) are representations of human tissues and provide organ-specific translational insights for physiological processes. In this study, a brain MPS was utilized to assess molecular signatures of neurotoxic and non-neurotoxic compounds using targeted and untargeted molecular approaches. The brain MPS comprising of human embryonic stem (ES) cell-derived neural progenitor cells seeded on three-dimensional (3D), chemically defined, polyethylene glycol hydrogels was treated with the neurotoxic drug, bortezomib and the non-neurotoxic drug, tamoxifen over 14-days. Possible toxic effects were monitored with human N-acetylaspartic acid (NAA) kinetics, which correlates the neuronal function/health and DJ-1/PARK7, an oxidative stress biomarker. Changes in NAA levels were observed as early as 2-days post-bortezomib treatment, while onset detection of oxidative stress (DJ-1) was delayed until 4-days post-treatment. Separately, the untargeted extracellular metabolomics approach revealed distinct fingerprints 2-days post-bortezomib treatment as perturbations in cysteine and glycerophospholipid metabolic pathways. These results suggest accumulation of reactive oxygen species associated with oxidative stress, and disruption of membrane structure and integrity. The NAA response was strongly correlated with changes in a subset of the detected metabolites at the same time point 2-days post-treatment. Moreover, these metabolite changes correlated strongly with DJ-1 levels measured at the later time point (4-days post-treatment). This suggests that early cellular metabolic dysfunction leads to later DJ-1 leakage and cell death, and that early measurement of this subset of metabolites could predict the later occurrence of cell death. While the approach demonstrated here provides an individual case study for proof of concept, we suggest that this approach can be extended for preclinical toxicity screening and biomarker discovery studies.
对人类药物不良反应进行早期评估对于避免长期伤害至关重要。然而，当前用于早期检测不良反应的方法仍缺乏预测性和器官特异性生物标志物来评估人类的不良反应。微生理系统（MPSs）是人体组织的模型，可为生理过程提供器官特异性的转化见解。在本研究中，利用脑MPS，采用靶向和非靶向分子方法评估神经毒性和非神经毒性化合物的分子特征。将由人胚胎干细胞（ES）衍生的神经祖细胞接种在三维（3D）、化学成分明确的聚乙二醇水凝胶上构建的脑MPS，用神经毒性药物硼替佐米和非神经毒性药物他莫昔芬处理14天。通过与人神经元功能/健康相关的人N-乙酰天门冬氨酸（NAA）动力学以及氧化应激生物标志物DJ-1/PARK7监测可能的毒性作用。硼替佐米治疗后2天就观察到NAA水平变化，而氧化应激（DJ-1）的起始检测延迟至治疗后4天。另外，非靶向细胞外代谢组学方法显示，硼替佐米治疗后2天，半胱氨酸和甘油磷脂代谢途径的扰动呈现出明显的指纹图谱。这些结果表明与氧化应激相关的活性氧积累以及膜结构和完整性的破坏。治疗后2天，NAA反应与同一时间点检测到的一部分代谢物的变化密切相关。此外，这些代谢物变化与后期（治疗后4天）测得的DJ-1水平密切相关。这表明早期细胞代谢功能障碍会导致后期DJ-1泄漏和细胞死亡，并且对这部分代谢物的早期测量可以预测后期细胞死亡的发生。虽然这里展示的方法提供了一个概念验证的个体案例研究，但我们认为这种方法可以扩展到临床前毒性筛查和生物标志物发现研究中。
