A1 polarized IPSCs-derived astrocytes with the PSEN1EΔ9 mutation show deregulated inflammatory dynamics

Abstract

Event Abstract Back to Event A1 polarized IPSCs-derived astrocytes with the PSEN1EΔ9 mutation show deregulated inflammatory dynamics Sofia Ferreira1, Gonçalo F. Garcia1, Sara Pinto1, Filipa Moreira1, Minna Oksanen2, Jari Koistinaho2, Adelaide Fernandes1, 3 and Dora Brites1, 3* 1 Research Institute for Medicines (iMed.ULisboa), Portugal 2 A.I. Virtanen Institute for Molecular Sciences, Faculty of Health Sciences, University of Eastern Finland, Finland 3 Faculdade de Farmácia, Universidade de Lisboa, Department of Biochemistry and Human Biology, Portugal Astrocytes with an A1 phenotype were identified in the prefrontal cortex of Alzheimer’s disease (AD) patients, and revealed to occur upon stimulation with specific inflammatory molecules released by activated microglia in an in vitro murine model (1). Differentiation of astrocytes from iPSCs generated from AD patients may overcome species differences and allow a better understanding of early alterations in AD pathogenesis. So, we investigated the phenotypes of astrocytes differentiated from AD-iPSCs, before and after A1-stimulation, focusing on the dynamics of inflammatory-associated mediators/miRNAs. iPSCs from patients with PSEN1ΔE9 mutation and controls were obtained under the JPND-MADGIC-project (2), and astroglial differentiation was performed as previously described (3). Cells were treated with TNF-α+IL-1α+C1q to induce A1 polarization (1) and small extracellular vesicles (sEVs) isolated by ultracentrifugation. Astrocyte characterization was accomplished by GFAP- and S100B-immunostaining, and detection of released sAPPβ by WB. Expression of pro-/anti-inflammatory markers and associated miRNAs was determined by qRT-PCR. Although almost 100% of astrocytes derived from iPSCs stained positive for S100B, less than 80% of AD ones were positive for GFAP. A1-activation of AD astrocytes further decreased GFAP+ cell number and led to increased sAPPβ secretion. No differences were observed in TNF-α and IL-10 gene expression between non-polarized control and AD astrocytes. However, A1-polarization elicited downregulation of TNF-α and IL-10 in control cells, in contrast with their marked upregulation in AD astrocytes. Further, AD non-polarized cells showed upregulated miR-21 and miR-125b vs. control astrocytes, while a marked downregulation occurred upon A1 polarization. Most attractively, these miRNAs were shown to be decreased in sEVs of AD non-polarized cells, but increased in AD astrocyte media, as well as in AD sEVs and media after A1 induction. Our data support the existence of deregulated inflammatory dynamics in iPSCs-derived astrocytes with the PSEN1EΔ9 mutation, and unveil miR-21 and miR-125b as potential biomarkers for AD. Acknowledgements Supported by EU-JPND research (grant agreement Nº643417 to DB) and by Fundação para a Ciência e a Tecnologia (JPco-fuND/0003/2015 to DB; UID/DTP/04138/2013 to iMed.ULIsboa and SFRH/BD/128738/2017 to GG). References 1- Liddelow SA. et al., Neurotoxic reactive astrocytes are induced by activated microglia, Nature. 2017; 541:481-487 2- http://www.cellmadgic.eu/p1 3- Oksanen M. et al., SEN1 Mutant iPSC-derived model reveals severe astrocyte pathology in Alzheimer's disease, Stem Cell Reports. 2017; 9:1885-1897. Keywords: Alzheimer’s disease, Astrocyte pathology, Inflammatory markers, Inflammatory-associated miRNAs, iPSC = induced pluripotent stem cell Conference: XVI Meeting of the Portuguese Society for Neuroscience (SPN2019), Lisboa, Portugal, 30 May - 1 Jun, 2019. Presentation Type: Poster presentation Topic: Glia / Neuroinflammation Citation: Ferreira S, Garcia GF, Pinto S, Moreira F, Oksanen M, Koistinaho J, Fernandes A and Brites D (2019). A1 polarized IPSCs-derived astrocytes with the PSEN1EΔ9 mutation show deregulated inflammatory dynamics. Front. Cell. Neurosci. Conference Abstract: XVI Meeting of the Portuguese Society for Neuroscience (SPN2019). doi: 10.3389/conf.fncel.2019.01.00025 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: 16 Apr 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 Sofia Ferreira Gonçalo F Garcia Sara Pinto Filipa Moreira Minna Oksanen Jari Koistinaho Adelaide Fernandes Dora Brites Google Sofia Ferreira Gonçalo F Garcia Sara Pinto Filipa Moreira Minna Oksanen Jari Koistinaho Adelaide Fernandes Dora Brites Google Scholar Sofia Ferreira Gonçalo F Garcia Sara Pinto Filipa Moreira Minna Oksanen Jari Koistinaho Adelaide Fernandes Dora Brites PubMed Sofia Ferreira Gonçalo F Garcia Sara Pinto Filipa Moreira Minna Oksanen Jari Koistinaho Adelaide Fernandes 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.