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Prediction and validation of regulatory role of microRNAs in zebrafish (Danio rerio) responses to nanoparticle exposure with in silico and in vitro toxicological approaches

Electronic Theses of LMU Munich (Ludwig-Maximilians-Universität München) 2019 Score: 30 ? 0–100 AI score estimating relevance to the microplastics field. Papers below 30 are filtered from public browse.
Moyan Hu, Moyan Hu

Summary

This thesis developed in silico and in vitro methods to predict how microRNAs regulate zebrafish responses to nanoparticle exposure, providing new tools for assessing nanoparticle toxicity. As nanoplastics are a type of nanoparticle, these methods have direct application to understanding how nanoplastics affect aquatic organisms at the molecular level.

Polymers
Body Systems
Study Type In vitro

The release of engineered nanoparticles as by-product of human activities in the environment can interfere with normal biology and health of the exposed organisms. MicroRNAs have been suggested as potential toxicology biomarkers, however the information about expression and role of microRNA in regulation of signaling pathways in organisms exposed to nanoparticles (NP) is limited. Summary of reported biological and pathological outcomes of NP induced toxicity in zebrafish was followed with in silico analysis of the genes potentially responsible for observed toxicological effects. After identifying relevant genes, we constructed six miRNA-mRNA regulatory networks involved in nanoparticle induced toxicological responses in zebrafish. Based on our prediction and selection criteria, we identified six miRNAs that overlapped in networks with high prediction scores, and were validated by previous mammalian and zebrafish microRNA profiling studies: dre-miR-124, -144, -148, -155, -19a, -223. As the next step, we validated the expression of these six miRNAs in THP-1 human monocytic cell line after the exposure to Polystyrene (PS NPs) and ARS labeled Titanium dioxide nanoparticles (TiO2-ARS NPs). Also, identification of miRNAs expression post exposure to PLGA nanoparticles and E. coli BioParticles was used to exclude potential activation and engagement of miRNAs through phagocytosis or pro-inflammatory specific responses. In our study, miR-155-5p showed the most promise as biomarker for PS NPs and TiO2-ARS NPs induced adverse effects. To determine potential for PS NPs and TiO2-ARS NPs for genotoxicity, time and dose dependent DNA damage profile induced by PS NPs or TiO2-ARS NPs was established by comet assay. Results indicated the severe DNA damage was triggered by both PS NPs and TiO2-ARS NPs. However, we observed that the expression of DNA damage repairing genes was elevated post TiO2-ARS NPs but not post PS NPs exposure, questioning the utility of the comet assay as universal assessment tool for genotoxicity induced by nanoparticles in general. It was observed that after PS NPs exposure the successful transfection of miR-155-5p mimic induced the expression of ATM, TAOK1, TRIP13, and APAF-1 while the expression of ERCC1 was attenuated. The ATM, APAF-1 and RAD51 were strongly activated post TiO2-ARS NPs stimulation in mimic-transfected cells. These observations suggest there is significant involvement of miR-155-5p in PS NPs and TiO2-ARS NPs induced adverse effects.

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