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While the exact disease mechanism remains unknown, previous studies suggest that mutant GFAP influences many cellular processes, including cytoskeleton stability, mechanosensing, metabolism, and proteasome function. While most studies have primarily focused on GFAP\u2010expressing astrocytes, GFAP is also expressed by radial glia and neural progenitor cells, prompting questions about the impact of GFAP mutations on central nervous system (CNS) development. In this study, we observed impaired differentiation of astrocytes and neurons in co\u2010cultures of astrocytes and neurons, as well as in neural organoids, both generated from AxD patient\u2010derived induced pluripotent stem (iPS) cells with a GFAP\n                    <jats:sup>R239C<\/jats:sup>\n                    mutation. Leveraging single\u2010cell RNA sequencing (scRNA\u2010seq), we identified distinct cell populations and transcriptomic differences between the mutant GFAP cultures and a corrected isogenic control. These findings were supported by results obtained with immunocytochemistry and proteomics. In co\u2010cultures, the GFAP\n                    <jats:sup>R239C<\/jats:sup>\n                    mutation resulted in an increased abundance of immature cells, while in unguided neural organoids and cortical organoids, we observed altered lineage commitment and reduced abundance of astrocytes. Gene expression analysis revealed increased stress susceptibility, cytoskeletal abnormalities, and altered extracellular matrix and cell\u2013cell communication patterns in the AxD cultures, which also exhibited higher cell death after stress. Overall, our results point to altered cell differentiation in AxD patient\u2010derived iPS\u2010cell models, opening new avenues for AxD research.\n                  <\/jats:p>","DOI":"10.1002\/glia.24618","type":"journal-article","created":{"date-parts":[[2024,9,23]],"date-time":"2024-09-23T05:18:43Z","timestamp":1727068723000},"page":"57-79","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Aberrant neurodevelopment in human\n                    <scp>iPS<\/scp>\n                    cell\u2010derived models of Alexander disease"],"prefix":"10.1002","volume":"73","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5014-5801","authenticated-orcid":false,"given":"Zuzana","family":"Matusova","sequence":"first","affiliation":[{"name":"Laboratory of Gene Expression Institute of Biotechnology of the Czech Academy of Sciences  Vestec Czechia"},{"name":"Faculty of Science Charles University  Prague Czechia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7663-4689","authenticated-orcid":false,"given":"Werner","family":"Dykstra","sequence":"additional","affiliation":[{"name":"Department of Translational Neuroscience University Medical Centre Utrecht Brain Centre, Utrecht University  Utrecht The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5208-2114","authenticated-orcid":false,"given":"Yolanda","family":"de Pablo","sequence":"additional","affiliation":[{"name":"Laboratory of Astrocyte Biology and CNS Regeneration, Center for Brain Repair, Department of Clinical Neuroscience Institute of Neuroscience and Physiology, Sahlgrenska Academy at the University of Gothenburg  Gothenburg Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1010-4287","authenticated-orcid":false,"given":"Oskar G.","family":"Zetterdahl","sequence":"additional","affiliation":[{"name":"Stem Cells, Aging and Neurodegeneration Lab, Department of Experimental Medical Science, Faculty of Medicine, Lund Stem Cell Center Lund University  Lund Sweden"},{"name":"Glial and Neuronal Biology Lab, Department of Experimental Medical Science, Faculty of Medicine, Lund Stem Cell Center Lund University  Lund Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2689-268X","authenticated-orcid":false,"given":"Isaac","family":"Canals","sequence":"additional","affiliation":[{"name":"Glial and Neuronal Biology Lab, Department of Experimental Medical Science, Faculty of Medicine, Lund Stem Cell Center Lund University  Lund Sweden"},{"name":"Division of Metabolism University Children's Hospital Zurich, University of Zurich  Zurich Switzerland"},{"name":"ITINERARE\u2013Innovative therapies in rare diseases, University Research Priority Program, University of Zurich  Zurich Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6110-0502","authenticated-orcid":false,"given":"Charlotte A. 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