iPSC-derived reactive astrocytes from patients with multiple sclerosis protect cocultured neurons in inflammatory conditions
Janis Kerkering, … , Marlen Alisch, Volker Siffrin
Janis Kerkering, … , Marlen Alisch, Volker Siffrin
Published May 23, 2023
Citation Information: J Clin Invest. 2023;133(13):e164637. https://doi.org/10.1172/JCI164637.
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Research Article Inflammation

iPSC-derived reactive astrocytes from patients with multiple sclerosis protect cocultured neurons in inflammatory conditions

Abstract

Multiple sclerosis (MS) is the most common chronic central nervous system inflammatory disease. Individual courses are highly variable, with complete remission in some patients and relentless progression in others. We generated induced pluripotent stem cells (iPSCs) to investigate possible mechanisms in benign MS (BMS), compared with progressive MS (PMS). We differentiated neurons and astrocytes that were then stressed with inflammatory cytokines typically associated with MS phenotypes. TNF-α/IL-17A treatment increased neurite damage in MS neurons from both clinical phenotypes. In contrast, TNF-α/IL-17A–reactive BMS astrocytes cultured with healthy control neurons exhibited less axonal damage compared with PMS astrocytes. Accordingly, single-cell transcriptomic BMS astrocyte analysis of cocultured neurons revealed upregulated neuronal resilience pathways; these astrocytes showed differential growth factor expression. Furthermore, supernatants from BMS astrocyte/neuronal cocultures rescued TNF-α/IL-17–induced neurite damage. This process was associated with a unique LIF and TGF-β1 growth factor expression, as induced by TNF-α/IL-17 and JAK-STAT activation. Our findings highlight a potential therapeutic role of modulation of astrocyte phenotypes, generating a neuroprotective milieu. Such effects could prevent permanent neuronal damage.

Authors

Janis Kerkering, Bakhrom Muinjonov, Kamil S. Rosiewicz, Sebastian Diecke, Charlotte Biese, Juliane Schiweck, Claudia Chien, Dario Zocholl, Thomas Conrad, Friedemann Paul, Marlen Alisch, Volker Siffrin

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Figure 5

Bulk transcriptome analysis of MS patient astrocytes from monocultures and cocultures with NGN2 neurons.

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Bulk transcriptome analysis of MS patient astrocytes from monocultures a...
Cells were treated for 24 hours with TNF-α/IL-17A or left untreated (control). Neurons were washed off, and remaining astrocytes were used for further processing. (A) PCA plot of bulk RNA-Seq of coculture-derived astrocytes shows distinct segregation of samples according to patient group (PC1) and according to cytokine treatment versus control (PC2). (B) Venn chart representing shared and unique genes comparing mono- and coculture-derived astrocytes. (C) Heatmap showing the top 25 regulated genes ranked by the adjusted P value. (D) Heatmap representing expression of genes encoding neurotrophic and JAK/STAT–related factors. The z score was calculated including all 12 samples. (E) Reverse transcriptase qPCR of selected targets confirmed differentially expressed genes between BMS and PMS after treatment. Statistical significance was tested with an unpaired 2-tailed t test; *P < 0.05, **P < 0.01, ***P < 0.001. (F and G) Enrichr analysis of activated pathways (F) and Gene Ontology (GO) labels of biological processes (G) comparing tCBMS versus tCPMS.