Zhang Tong, Gaffrey Matthew J, Thomas Dennis G, Weber Thomas J, Hess Becky M, Weitz Karl K, Piehowski Paul D, Petyuk Vladislav A, Moore Ronald J, Qian Wei-Jun, Thrall Brian D
Biological Sciences Division, Pacific Northwest National Laboratory, Richland WA 99352.
Signatures Sciences and Technology Division, Pacific Northwest National Laboratory, Richland, WA 99352.
NanoImpact. 2020 Jan;17. doi: 10.1016/j.impact.2019.100194. Epub 2019 Nov 23.
Responsible implementation of engineered nanomaterials (ENMs) into commercial applications is an important societal issue, driving demand for new approaches for rapid and comprehensive evaluation of their bioactivity and safety. An essential part of any research focused on identifying potential hazards of ENMs is the appropriate selection of biological endpoints to evaluate. Herein, we use a tiered strategy employing both targeted biological assays and untargeted quantitative proteomics to elucidate the biological responses of human THP-1 derived macrophages across a library of metal/metal oxide ENMs, raised as priority ENMs for investigation by NIEHS's Nanomaterial Health Implications Research (NHIR) program. Our results show that quantitative cellular proteome profiles readily distinguish ENM types based on their cytotoxic potential according to induction of biological processes and pathways involved in the cellular antioxidant response, TCA cycle, oxidative stress, endoplasmic reticulum stress, and immune responses as major processes impacted. Interestingly, bioinformatics analysis of differentially expressed proteins also revealed new biological processes that were influenced by all ENMs independent of their cytotoxic potential. These included biological processes that were previously implicated as mechanisms cells employ as adaptive responses to low levels of oxidative stress, including cell adhesion, protein translation and protein targeting. Unsupervised clustering revealed the most striking proteome changes that differentiated ENM classes highlight a small subset of proteins involved in the oxidative stress response (HMOX1), protein chaperone functions (HS71B, DNJB1), and autophagy (SQSTM), providing a potential new panel of markers of ENM-induced cellular stress. To our knowledge, the results represent the most comprehensive profiling of the biological responses to a library of ENMs conducted using quantitative mass spectrometry-based proteomics. The results provide a basis to identify the patterns of a diverse set of cellular pathways and biological processes impacted by ENM exposure in an important immune cell type, laying the foundation for multivariate, pathway-level structure activity assessments of ENMs in the future.
将工程纳米材料（ENMs）负责任地应用于商业领域是一个重要的社会问题，这推动了对快速全面评估其生物活性和安全性的新方法的需求。任何旨在识别ENMs潜在危害的研究的一个重要部分是选择合适的生物学终点进行评估。在此，我们采用一种分层策略，结合靶向生物学检测和非靶向定量蛋白质组学，以阐明人类THP-1衍生巨噬细胞对一系列金属/金属氧化物ENMs的生物学反应，这些ENMs是美国国立环境卫生科学研究所（NIEHS）的纳米材料健康影响研究（NHIR）项目列为优先研究对象的。我们的结果表明，定量细胞蛋白质组图谱能够根据细胞抗氧化反应、三羧酸循环、氧化应激、内质网应激和免疫反应等生物学过程和途径的诱导情况，根据其细胞毒性潜力轻松区分ENM类型，这些是受影响的主要过程。有趣的是，对差异表达蛋白质的生物信息学分析还揭示了所有ENMs都能影响的新生物学过程，而与它们的细胞毒性潜力无关。这些过程包括以前被认为是细胞对低水平氧化应激的适应性反应机制的生物学过程，如细胞黏附、蛋白质翻译和蛋白质靶向。无监督聚类揭示了区分ENM类别的最显著蛋白质组变化，突出了一小部分参与氧化应激反应（HMOX1）、蛋白质伴侣功能（HS71B、DNJB1）和自噬（SQSTM）的蛋白质，提供了一组潜在的ENM诱导细胞应激的新标志物。据我们所知，这些结果代表了使用基于定量质谱的蛋白质组学对一系列ENMs的生物学反应进行的最全面分析。这些结果为识别重要免疫细胞类型中受ENM暴露影响的各种细胞途径和生物学过程的模式提供了基础，为未来对ENMs进行多变量、途径水平的结构活性评估奠定了基础。
