Targeting astrocyte and motor neuron specific miRNAs to prevent neuro-immune dysregulation in ALS

Event Abstract Back to Event Targeting astrocyte and motor neuron specific miRNAs to prevent neuro-immune dysregulation in ALS Ana R. Vaz1, 2, Catarina Sequeira1, Daniela Vizinha1, Marta Barbosa1, Cátia Gomes1 and Dora Brites1, 2, 3* 1 Research Institute for Medicines (iMed.ULisboa), Portugal 2 Faculdade de Farmácia, Universidade de Lisboa, Department of Biochemistry and Human Biology, Portugal 3 Faculdade de Farmácia, Universidade de Lisboa, Portugal Amyotrophic lateral sclerosis (ALS) is characterized by motor neurons (MNs) loss and glial reactivity in brain and spinal cord (SC). A dysregulated expression of microRNAs (miRNAs) have been implicated in neurodegeneration and neuroinflammation in ALS. MiRNA(miR)-124 is elevated in the cerebrospinal fluid of ALS patients and in the SC of the ALS mice with the G93A mutation in human superoxide dismutase-1 (mSOD1). We demonstrated that this miRNA is upregulated in MN-NSC-34 overexpressing mSOD1 and in small extracellular vesicles isolated from cell secretome [1]. We also observed miR-155 upregulation in the SC of pe-symptomatic mSOD1 mice [2], often associated with mSOD1 microglia [3], but with unknown effect when expressed by astrocytes. In contrast, miR-146a was downregulated either in the cortical brain (CB) of symptomatic mSOD1 mice and in isolated astrocytes [4], suggesting distinct miRNA deregulation depending on the CNS region. Here, we evaluated the effects of miR-155/146a/124 modulation in astrocytic and MN function. For that, we used primary astrocytes from mSOD1 mice pups (CB or SC) and mSOD1 MN-NSC-34. Cells were transfected with anti-miR-155 (SC astrocytes), premiR-146a (CB astrocytes) or pre/anti-miR-124 (MN). We also explored whether changes in MN-miR-124 expression impact on secretome-mediated astrocytic function. Downregulation of miR-155 in mSOD1 SC astrocytes counteracted the inflammatory milieu, by decreasing TNF-α/IL-1β/HMGB1/miR-124, and increasing IL-6/IL-10/SOCS1/miR-146a expression, without changing the astrocytic aberrant phenotype (GFAPlow/Cx43high/S100Bhigh). However, miR-146a upregulation in mSOD1 CB astrocytes neutralized both the phenotypic aberrancy and the inflammatory profile. The upregulation of miR-124 in WT-MNs mimicked the pathological features of mSOD1-MNs, namely inflammatory-associated-miRNAs (increased miR-125b/decreased miR-146a/-21), synaptic dysfunction (increased synaptophysin/decreased PDS-95), neurite loss, reduced mitochondria viability and axonal transport deficits (increased dynein/decreased kinesin), while miR-124 downregulation was able to prevent those dysfunctionalities in mSOD1-MNs. The astrocytic aberrant phenotype of mSOD1 SC astrocytes was observed in WT cells upon incubation with secretome from mSOD1-MNs or pre-miR-124-WT-MN, but prevented in the presence of the secretome of anti-miR-124-mSOD1-MN. More importantly, secretome derived from anti-miR-124-mSOD1-MN rescued the aberrant phenotype of mSOD1 SC astrocytes by increasing GFAP/GLT-1 expression. Overall, our results highlight that combined modulation of specific miRNAs in different target cells may represent a promising therapeutic potential in ALS disease. Acknowledgements Funded by SCML (ALS Research Grant ELA-2015-002) and FCT (PTDC/MED-NEU/31395/2017 and LISBOA-01-0145-FEDER-031395 to DB, UID/DTP/04138/2013 to iMed.ULisboa, and fellowship ARV-SFRH/BPD/76590/2011). References [1] Pinto et al Front Neurosci. 2017 [2] Cunha et al Mol Neurobiol. 2018 [3] Butovsky el al Ann Neurol. 2015 [4] Gomes et al Mol Neurobiol. 2018 Keywords: . miRNA modulation, mSOD1 MN-like NSC-34 cells, astrocytic activation and phenotype, Secretome, regionalized astrocytes miRNA signature Conference: XVI Meeting of the Portuguese Society for Neuroscience (SPN2019), Lisboa, Portugal, 30 May - 1 Jun, 2019. Presentation Type: Oral presentation Topic: Glia / Neuroinflammation Citation: Vaz AR, Sequeira C, Vizinha D, Barbosa M, Gomes C and Brites D (2019). Targeting astrocyte and motor neuron specific miRNAs to prevent neuro-immune dysregulation in ALS. Front. Cell. Neurosci. Conference Abstract: XVI Meeting of the Portuguese Society for Neuroscience (SPN2019). doi: 10.3389/conf.fncel.2019.01.00049 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 28 Feb 2019; Published Online: 27 Sep 2019. * Correspondence: Prof. Dora Brites, Research Institute for Medicines (iMed.ULisboa), Lisboa, Portugal, dbrites@ff.ulisboa.pt Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Ana R Vaz Catarina Sequeira Daniela Vizinha Marta Barbosa Cátia Gomes Dora Brites Google Ana R Vaz Catarina Sequeira Daniela Vizinha Marta Barbosa Cátia Gomes Dora Brites Google Scholar Ana R Vaz Catarina Sequeira Daniela Vizinha Marta Barbosa Cátia Gomes Dora Brites PubMed Ana R Vaz Catarina Sequeira Daniela Vizinha Marta Barbosa Cátia Gomes Dora Brites Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.