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 6

Supernatant analysis of MS patient astrocyte/NGN2-neuron cocultures.

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Supernatant analysis of MS patient astrocyte/NGN2-neuron cocultures. (A)...
(A) Supernatants of TNF-α/IL-17A–treated and untreated NGN2 and BMS astrocytes were collected after 24 hours of stimulation and applied on monocultures of TNF-α/IL-17A–preincubated NGN2-neurons. (B) NGN2-neurons were treated with TNF-α/IL-17A (50 ng/mL) for 24 hours (represented by “+”), and medium was replaced with untreated or treated coculture supernatants. Each data point represents a microscopic field of view (641 × 479 μm) of 3 independent experiments depicted by different symbols; pooled data show the mean from 3 patients (different colors) and 3 independent experiments (different symbols). Neurons showed protection when given supernatants of treated BMS cocultures. “+” represents an arbitrary proportion of the target in tBMS compared with tPMS; “=” indicates an equal level. (C and D) Bead-based multiplex assay and ELISA (CTGF) analysis of supernatants. LIF, BDNF, and TGF-β1 concentrations were significantly higher in tCBMS than in tCPMS. Each data point represents 1 sample collectively of 3 individual patients (different colors) and 3 independent experiments (different symbols). Statistical significance was tested with a Kruskal-Wallis test; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.