Immune response of BV-2 microglial cells is impacted by peroxisomal beta-oxidation

Ali Tawbeh; Quentin Raas; Mounia Tahri-Joutey; Céline Keime; Romain Kaiser; Doriane Trompier; Boubker Nasser; Emma Bellanger; Marie Dessard; Yannick Hamon; Alexandre Benani; Francesca Di Cara; Tânia Cunha Alves; Johannes Berger; Isabelle Weinhofer; Stéphane Mandard; Mustapha Cherkaoui-Malki; Pierre Andreoletti; Catherine Gondcaille; Stéphane Savary

doi:10.3389/fnmol.2023.1299314

Immune response of BV-2 microglial cells is impacted by peroxisomal beta-oxidation

Front Mol Neurosci. 2023 Dec 18:16:1299314. doi: 10.3389/fnmol.2023.1299314. eCollection 2023.

Authors

Ali Tawbeh^#¹, Quentin Raas^#¹, Mounia Tahri-Joutey^{1

2}, Céline Keime³, Romain Kaiser³, Doriane Trompier¹, Boubker Nasser², Emma Bellanger⁴, Marie Dessard⁴, Yannick Hamon⁴, Alexandre Benani⁵, Francesca Di Cara⁶, Tânia Cunha Alves⁷, Johannes Berger⁷, Isabelle Weinhofer⁷, Stéphane Mandard⁸, Mustapha Cherkaoui-Malki¹, Pierre Andreoletti¹, Catherine Gondcaille¹, Stéphane Savary¹

Affiliations

¹ Laboratoire Bio-PeroxIL EA7270, University of Bourgogne, Dijon, France.
² Laboratory of Biochemistry, Neurosciences, Natural Resources and Environment, Faculty of Sciences and Techniques, University Hassan I, Settat, Morocco.
³ Plateforme GenomEast, IGBMC, CNRS UMR 7104, Inserm U1258, University of Strasbourg, Illkirch, France.
⁴ Aix Marseille Univ, CNRS, INSERM, CIML, Marseille, France.
⁵ Centre des Sciences du Goût et de l'Alimentation, CNRS, INRAE, Institut Agro, University of Bourgogne, Dijon, France.
⁶ Department of Microbiology and Immunology, Dalhousie University, IWK Health Centre, Halifax, NS, Canada.
⁷ Department of Pathobiology of the Nervous System, Center for Brain Research, Medical University of Vienna, Vienna, Austria.
⁸ LipSTIC LabEx, University of Bourgogne, INSERM LNC UMR1231, Dijon, France.

^# Contributed equally.

Abstract

Microglia are crucial for brain homeostasis, and dysfunction of these cells is a key driver in most neurodegenerative diseases, including peroxisomal leukodystrophies. In X-linked adrenoleukodystrophy (X-ALD), a neuroinflammatory disorder, very long-chain fatty acid (VLCFA) accumulation due to impaired degradation within peroxisomes results in microglial defects, but the underlying mechanisms remain unclear. Using CRISPR/Cas9 gene editing of key genes in peroxisomal VLCFA breakdown (Abcd1, Abcd2, and Acox1), we recently established easily accessible microglial BV-2 cell models to study the impact of dysfunctional peroxisomal β-oxidation and revealed a disease-associated microglial-like signature in these cell lines. Transcriptomic analysis suggested consequences on the immune response. To clarify how impaired lipid degradation impacts the immune function of microglia, we here used RNA-sequencing and functional assays related to the immune response to compare wild-type and mutant BV-2 cell lines under basal conditions and upon pro-inflammatory lipopolysaccharide (LPS) activation. A majority of genes encoding proinflammatory cytokines, as well as genes involved in phagocytosis, antigen presentation, and co-stimulation of T lymphocytes, were found differentially overexpressed. The transcriptomic alterations were reflected by altered phagocytic capacity, inflammasome activation, increased release of inflammatory cytokines, including TNF, and upregulated response of T lymphocytes primed by mutant BV-2 cells presenting peptides. Together, the present study shows that peroxisomal β-oxidation defects resulting in lipid alterations, including VLCFA accumulation, directly reprogram the main cellular functions of microglia. The elucidation of this link between lipid metabolism and the immune response of microglia will help to better understand the pathogenesis of peroxisomal leukodystrophies.

Keywords: adrenoleukodystrophy; antigen presentation; immune response; inflammation; microglia; peroxisome; phagocytosis.

Copyright © 2023 Tawbeh, Raas, Tahri-Joutey, Keime, Kaiser, Trompier, Nasser, Bellanger, Dessard, Hamon, Benani, Di Cara, Cunha Alves, Berger, Weinhofer, Mandard, Cherkaoui-Malki, Andreoletti, Gondcaille and Savary.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. The authors warmly acknowledge the Fondation Maladies Rares, which supported our transcriptomic analysis project (GenOmics: High throughput sequencing and rare diseases, call 2017-20170615). Sequencing was performed by the GenomEast platform, a member of the France Génomique consortium (ANR-10-INBS-0009). The authors would like to acknowledge the Canadian New Frontiers Research Funds Exploration (NFRF-E 19-00007) and the regional council of Bourgogne Franche-Comté (Project PERSIL 2019) for their support. We are also grateful for the networking support by the COST Action CA 16,112 NutRedOx (Personalized Nutrition in aging society: redox control of major age-related diseases), supported by COST (European Cooperation in Science and Technology). This study was supported by institutional grants from INSERM, CNRS, and Aix-Marseille University to the CIML and a program grant from the French National Research Agency (ANR-17-CE15-0032). The project leading to this publication has received funding from the Excellence Initiative of Aix-Marseille University—A^*MIDEX, a French Investissements d'Avenir program. Part of this research was also funded by grants from the Institut National de la Santé et de la Recherche Médicale (INSERM) and by a French Government grant managed by the French National Research Agency under the program “Investissements d'Avenir” with reference ANR-11 LABX-0021 (Lipstic Labex). The laboratory BioPeroxIL was funded by the Ministère de l'Education Nationale, de l'Enseignement Supérieur et de la Recherche (France), and by the University of Bourgogne. MT-J was funded by CNRST (PhD excellence grant number: 17UHP2019, Morocco) and by the Action Intégrée of the Comité Mixte Inter-Universitaire Franco-Marocain (no. TBK 19/92 no. Campus France: 41501RJ) from the PHC Toubkal program, Ministère des Affaires Étrangères. IW was supported by the Austrian Science Fund KLI 837-B and by the European Leukodystrophy Association (ELA) Germany.