Inflammatory transcriptomic signatures in a human cellular NMOSD model reveal upregulation of NF-?B and IL6 pathways

Neuromyelitis optica spectrum disorder (NMOSD) is a rare neurological autoimmune disease caused autoantibodies targeting the astrocytic water channel aquaporin-4 (AQP4). Binding to AQP4 initiates the activation of innate immune components, especially the complement system. Both in-vivo and in-vitro models have been developed to study the molecular pathophysiology of NMOSD. The aim of our study was to characterize the molecular response of four human cell lines (AQP4-ECFP expressing U-87MG glioblastoma cells, U-87MG expressing only ECFP, HEK293 cells expressing AQP4-EmGFP and human primary astrocytes) to a treatment with AQP4 antibody E5415A and human complement. Complement-dependent cytotoxicity was induced by this treatment in AQP4-expressing cells by the classical complement pathway. Transcriptomic profiles of the in-vitro U-87MG-AQP4-ECFP model and an in-vivo rat model shared a proinflammatory shift towards NF-?B and interleukin-6 pathways. These findings were confirmed on the mRNA and protein levels and treatment with serum samples from AQP4 antibody seropositive NMOSD patients resulted in a similar response. Additionally, NF-?B upregulation was shown by immunohistochemistry in medulla oblongata lesions of NMOSD patients. In conclusion, interleukin-6 and NF-?B pathways play a key role in inflammation caused by the activation of the classical complement pathway in a human cellular model of NMOSD using U-87MG-AQP4-ECFP cells. Brandl, S., Yu, Q., Hagenbuchner, J. et al. Inflammatory transcriptomic signatures in a human cellular NMOSD model reveal upregulation of NF-?B and IL6 pathways. Sci Rep 15, 43346 (2025). https://doi.org/10.1038/s41598-025-27335-9 Overall design: RNA-seq profiling of U-87MG-AQP4-ECFP, U-87MG-ECFP, HA and HEK293-AQP4-EmGFP cells treated with anti-AQP4 antibody E5415A and active and heat-inactivated human complement *************************************************************** Submitter states that only one of the two paired-end raw files is provided for the following samples: S10, S11, S13, S14 and S16 and no raw files are provided S15, due to file loss. ***************************************************************