20.500.12556/DiRROS-21158 Transcriptomic and proteomic profiling of bi-partite and tri-partite murine iPSC-derived neurospheroids under steady-state and inflammatory condition Induced-pluripotent stem cell (iPSC)-derived neurospheroid (NSPH) models are an emerging in vitro toolkit to study the influence of inflammatory triggers on neurodegeneration and repair in a 3D neural environment. In contrast to their human counterpart, the absence of murine iPSC-derived NSPHs for profound characterisation and validation studies is a major experimental research gap, even though they offer the only possibility to truly compare or validate in vitro NSPH responses with in vivo brain responses. To contribute to these developments, we here describe the generation and characterisation of 5-week-old CX3CR1eGFP+/- CCR2RFP+/- murine (m)iPSC-derived bi-partite (neurons + astrocytes) and tri-partite (neurons + astrocytes + microglia) NSPH models that can be subjected to cellular activation following pro-inflammatory stimulation. First, cytokine analysis demonstrates that both bi-partite and tri-partite NSPHs can be triggered to release IL6 and CXCL10 following three days of stimulation with, respectively, TNFα + IL1β + IFNγ and LPS + IFNγ. Additionally, immunocytochemical analysis for G3BP1 and PABPC1 revealed the development of stress granules in both bi-partite and tri-partite NSPHs after 3 days of stimulation. To further investigate the observed signs of inflammatory response and cellular stress, we performed an untargeted transcriptomic and proteomic analysis of bi- and tri-partite NSPHs under steady-state and inflammatory conditions. Here, using the combined differential gene and protein expression profiles between unstimulated and stimulated NSPHs, Ingenuity Pathway Analysis (IPA) confirms the activation of canonical pathways associated with inflammation and cellular stress in both bi-partite and tri-partite NSPHs. Moreover, our multi-omics analysis suggests a higher level of downstream inflammatory responses, impairment of homeostatic and developmental processes, as well as activation of cell death processes in stimulated tri-partite NSPHs compared to bi-partite NSPHs. Concluding, these results emphasise the advantages of including microglia in NSPH research to study inflammation-induced neurodegeneration in a 3D neural environment. murine induced-pluripotent cells neurospheroids inflammatory response stress granules transcriptomics proteomics vnetni odziv proteomika true false true Elsevier Nizozemska Angleški jezik Slovenski jezik © 2024 Elsevier Inc. All rights are reserved, including those for text and data mining, AI training, and similar technologies. Neznano 2025-01-08 13:26:39 2025-01-08 13:26:40 2025-01-09 03:38:02 0000-00-00 00:00:00 2024 0 0 Nasl. z nasl. zaslona; Soavtorja iz Slovenije: Helena Motaln, Boris Rogelj; Opis vira z dne 2. 9. 2024; str. 1-12 Vol. 121 Oct. 2024 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 2024-05-24 2024-07-06 2024-07-14 57 1090-2139 10.1016/j.bbi.2024.07.008 206077443 https://www.sciencedirect.com/science/article/pii/S0889159124004756?via%3Dihub 1 d07a1b59-cdbb-11ef-ba3f-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=30017 RAZ_Di_Stefano_Julia_2024.pdf RAZ_Di_Stefano_Julia_2024.pdf 1 BB60590C9445DC51CC8735372AAB1566 7b224af68251c868ed047c5830da790d592bde90d6162b13e3fd28d60dbe0793 22f4cd5a-cdbd-11ef-ba3f-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=30018 Institut Jožef Stefan 0 0 0