TNF-alpha-induced microglia activation requires miR-342: impact on NF-kB signaling and neurotoxicity
| Autor: | Mário A. Barbosa, Teresa Summavielle, Susana G. Santos, Jaime Freitas, Joana Bravo, Maria Inês Almeida, Joao Paulo Bras |
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| Přispěvatelé: | Instituto de Investigação e Inovação em Saúde, Repositório Científico do Instituto Politécnico do Porto |
| Rok vydání: | 2020 |
| Předmět: |
Cancer Research
Gene Expression Regulation / drug effects Neurotoxins / toxicity Neuroimmunology Inflammation / genetics Tumor Necrosis Factor-alpha / pharmacology Inflammation / pathology Primary microglia Microglia lcsh:Cytology Chemistry Enzyme-linked immunosorbent assay (ELISA) NF-kappa B MicroRNAs / metabolism Cell biology DNA-Binding Proteins medicine.anatomical_structure Microglia / metabolism miRNAs Cytokines Tumor necrosis factor alpha Neurotoxins Immunology Mixed glial cells Context (language use) Models Biological Article NF-kappa B / metabolism Cell Line Cellular and Molecular Neuroscience Mediator Downregulation and upregulation medicine Animals Rats Wistar lcsh:QH573-671 Neuroinflammation Inflammation Microglia / pathology Transcription Factors / metabolism Tumor Necrosis Factor-alpha Neurotoxicity N9 microglial cells DNA-Binding Proteins / metabolism Cell Biology Microglia / drug effects medicine.disease miR-342 Mice Inbred C57BL MicroRNAs Cytokines / metabolism MicroRNAs / genetics Animals Newborn Gene Expression Regulation nervous system TNF-alpha-induced microglia activation Neuron Transcription Factors |
| Zdroj: | Cell Death and Disease, Vol 11, Iss 6, Pp 1-15 (2020) Cell Death & Disease Repositório Científico de Acesso Aberto de Portugal Repositório Científico de Acesso Aberto de Portugal (RCAAP) instacron:RCAAP |
| ISSN: | 2041-4889 |
| DOI: | 10.1038/s41419-020-2626-6 |
| Popis: | Growing evidences suggest that sustained neuroinflammation, caused by microglia overactivation, is implicated in the development and aggravation of several neurological and psychiatric disorders. In some pathological conditions, microglia produce increased levels of cytotoxic and inflammatory mediators, such as tumor necrosis factor alpha (TNF-α), which can reactivate microglia in a positive feedback mechanism. However, specific molecular mediators that can be effectively targeted to control TNF-α-mediated microglia overactivation, are yet to be uncovered. In this context, we aim to identify novel TNF-α-mediated micro(mi)RNAs and to dissect their roles in microglia activation, as well as to explore their impact on the cellular communication with neurons. A miRNA microarray, followed by RT-qPCR validation, was performed on TNF-α-stimulated primary rat microglia. Gain- and loss-of-function in vitro assays and proteomic analysis were used to dissect the role of miR-342 in microglia activation. Co-cultures of microglia with hippocampal neurons, using a microfluidic system, were performed to understand the impact on neurotoxicity. Stimulation of primary rat microglia with TNF-α led to an upregulation of Nos2, Tnf, and Il1b mRNAs. In addition, ph-NF-kB p65 levels were also increased. miRNA microarray analysis followed by RT-qPCR validation revealed that TNF-α stimulation induced the upregulation of miR-342. Interestingly, miR-342 overexpression in N9 microglia was sufficient to activate the NF-kB pathway by inhibiting BAG-1, leading to increased secretion of TNF-α and IL-1β. Conversely, miR-342 inhibition led to a strong decrease in the levels of these cytokines after TNF-α activation. In fact, both TNF-α-stimulated and miR-342-overexpressing microglia drastically affected neuron viability. Remarkably, increased levels of nitrites were detected in the supernatants of these co-cultures. Globally, our findings show that miR-342 is a crucial mediator of TNF-α-mediated microglia activation and a potential target to tackle microglia-driven neuroinflammation. We would like to thank Dr. João Relvas laboratory for the help with N9 microglia cell culture; Dr. Sofia Lamas for the guidance on the animal welfare and support with animal experiments (Animal facility, i3S); and to LC Sciences for the miRNA microarray data and analysis. The mass spectrometry technique was performed by Hugo Osório at the i3S Proteomics Scientific Platform with support from the Portuguese Mass Spectrometry Network, integrated in the National Roadmap of Research Infrastructures of Strategic Relevance (ROTEIRO/0028/2013; LISBOA-01–0145-FEDER-022125). This work was funded by project NORTE-01–0145-FEDER-000012, supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). J.P.B. and J.B. are supported by FCT–Fundação para a Ciência e Tecnologia, through BiotechHealth PhD program fellowship (PD/BD/135490/2018) and Areas of Basic and Applied Biology PhD program fellowship (PD/BD/135450/2017), respectively. |
| Databáze: | OpenAIRE |
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