High-content analysis for mitophagy response to nanoparticles: A potential sensitive biomarker for nanosafety assessment |
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| Authors: | Chengyong He Shengwei Jiang Huan Yao Liyin Zhang Chuanli Yang Shan Jiang Fengkai Ruan Denglin Zhan Gang Liu Zhongning Lin Yuchun Lin Xiaoyuan Chen |
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| Institution: | 1. State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Public Health, Xiamen University, Xiamen, Fujian, PR China;2. Laboratory of Molecular Imaging and Nanomedicine, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, United States |
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| Abstract: | Mitophagy, a selective autophagy of mitochondria, clears up damaged mitochondria to maintain cell homeostasis. We performed high-content analysis (HCA) to detect the increase of PINK1, an essential protein controlling mitophagy, in hepatic cells treated with several nanoparticles (NPs). PINK1 immunofluorescence-based HCA was more sensitive than assays and detections for cell viability and mitochondrial functions. Of which, superparamagnetic iron oxide (SPIO)-NPs or graphene oxide-quantum dots (GO-QDs) was selected as representatives for positive or negative inducer of mitophagy. SPIO-NPs, but not GO-QDs, activated PINK1-dependent mitophagy as demonstrated by recruitment of PARKIN to mitochondria and degradation of injured mitochondria. SPIO-NPs caused the loss of mitochondrial membrane potential, decrease in ATP, and increase in mitochondrial reactive oxide species and Ca2+. Blocking mitophagy with PARKIN siRNA aggravated the cytotoxicity of SPIO-NPs. Taken together, PINK1 immunofluorescence-based HCA is considered to be an early, sensitive, and reliable approach to evaluate the bioimpacts of NPs. |
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| Keywords: | Mitophagy Mitochondrial quality High-content analysis Nanosafety assessment Superparamagnetic iron oxide nanoparticles Graphene oxide-quantum dots Au gold 2+ CdTe cadmium telluride 2 cerium dioxide DAPI 4’ 6-diamidino-2-phenylindole DLS dynamic light scattering GO graphene oxide HCA high-content analysis mitoROS mitochondrial reactive oxide species MMP mitochondrial membrane potential MOM mitochondrial outer membrane MRI magnetic resonance imaging NMs nanomaterials NPs nanoparticles PBS phosphate buffered solution PINK1 PTEN-induced putative kinase 1 PLA proximity ligation assay QDs quantum dots ROS reactive oxide species 2 silicon dioxide siRNA small interfering RNA SPIO superparamagnetic iron oxide TEM transmission electron microscopy ZnO zinc oxide |
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