Menevse, A. N.; Hussein, A.; Sax, J.; Sorrentino, A.; Volpin, V.; Khandelwal, N.; Stamova, S.; Muehlbauer, J.; Schuch, N.; Ott-Roetzer, B.; Engelhorn, A.; Linder, H.; Rathinasamy, A.; Chen, C.-Y.; Sanna, F. C.; Wurzel, A.; Bellersheim, L.; Xydia, M.; Lohmayer, R.; Tuemen, D.; Guelow, K.; Michels, T.; Beckhove, P.

Multiple sclerosis (MS) is a progressive neuroinflammatory demyelinating disease of the central nervous system (CNS) that remains incurable. Autoreactive myelin-specific T cells contribute to immunopathology by directly targeting and damaging oligodendrocytes in situ. In oligodendrocytes, several immune-modulatory functions have been described that can ameliorate immune damage. However, a systematic discovery of cell-intrinsic mechanisms that protect oligodendrocytes against T cell-derived cytotoxic mechanisms has not been performed. We used human MO3.13 oligodendrocytic cells and human antigen-specific cytotoxic T cells to conduct a high-throughput (HTP) RNAi-based screen with altogether 4155 genes to identify oligodendrocyte-intrinsic immune resistance genes (IRGs). The screen revealed 133 candidate IRGs. Among them, we validated 32, which exerted a strong immuno-protective phenotype. We studied IRG expression landscapes in human brain cell subsets from postmortem brain tissues of MS and control individuals. This revealed clustered expression of IRGs in a cell-type and oligodendrocyte subset-specific manner and differential IRG expression between MS patients and controls in distinct oligodendrocyte subclusters. ChEA3 analysis revealed cell type-specific expression of transcription factors that can drive expression of respective IRGs. Explorative molecular mode of action analyses of five selected IRGs, STK11, KCNH8, ABCA2, SLC1A3 and CHRNA1 revealed that these prevented death receptor-mediated apoptosis induced by T cell-derived cytotoxic molecules. In particular, they controlled TRAIL-induced apoptosis by suppressing JNK1 activation through interfering with several upstream pathways regulating metabolic, potassium, cholesterol, glutamate and acetylcholine homeostasis. In addition, STK11, ABCA2, and CHRNA1 regulated TRAIL-R2 surface expression contributing to increased TRAIL-sensitivity whereas KCNH8 expression in oligodendrocytes inhibited secretion of inflammatory cytokines by cytotoxic T cells. Taken together, we here demonstrate the existence of multiple co-expressed IRGs in human oligodendrocytes that regulate multifaceted mechanisms of T cell resistance and are dysregulated in oligodendrocyte subsets of MS patients.