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High-content analysis for mitophagy response to nanoparticles: A potential sensitive biomarker for nanosafety assessment

Chengyong He,Shengwei Jiang,Huan Yao,Liyin Zhang,Chuanli Yang,Shan Jiang,Fengkai Ruan,Denglin Zhan,Gang Liu,Zhongning Lin,Yuchun Lin,Xiaoyuan Chen

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 Ca 2+ . 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.