IFN-α Induces Heterogenous ROS Production in Human β-Cells

Wagner, L. E.; Melnyk, O.; Turner, A.; Duffett, B. E.; Muralidharan, C.; Martinez-Irizarry, M. M.; Arvin, M. C.; Orr, K. S.; Manduchi, E.; Kaestner, K. H.; Brozinick, J. T.; Linnemann, A. K.

doi:10.1101/2025.02.19.639120

bioRxiv · 10.1101/2025.02.19.639120

IFN-α Induces Heterogenous ROS Production in Human β-Cells

Wagner, L. E.·Melnyk, O.·Turner, A.·Duffett, B. E.·Muralidharan, C.·Martinez-Irizarry, M. M.·Arvin, M. C.·Orr, K. S.·Manduchi, E.·Kaestner, K. H.·Brozinick, J. T.·Linnemann, A. K.

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Abstract

Type 1 diabetes (T1D) is a multifactorial disease involving genetic and environmental factors, including viral infection. We investigated the impact of interferon alpha (IFN-), a cytokine produced during the immune response to viral infection or the presence of un-edited endogenous double-stranded RNAs, on human {beta}-cell physiology. Intravital microscopy on transplanted human islets using a {beta}-cell-selective reactive oxygen species (ROS) biosensor (RIP1-GRX1-roGFP2), revealed a subset of human {beta}-cells that acutely produce ROS in response to IFN-. Comparison to Integrated Islet Distribution Program (IIDP) phenotypic data revealed that healthier donors had more ROS accumulating cells. In vitro IFN- treatment of human islets similarly elicited a heterogenous increase in superoxide production that originated in the mitochondria. To determine the unique molecular signature predisposing cells to IFN- stimulated ROS production, we flow sorted human islets treated with IFN-. RNA sequencing identified genes involved in inflammatory and immune response in the ROS-producing cells. Comparison with single cell RNA-Seq datasets available through the Human Pancreas Analysis Program (HPAP) showed that genes upregulated in ROS-producing cells are enriched in control {beta}-cells rather than T1D donors. Combined, these data suggest that IFN- stimulates mitochondrial ROS production in healthy human {beta}-cells, potentially predicting a more efficient antiviral response.

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Wagner, L. E., Melnyk, O., Turner, A., Duffett, B. E., Muralidharan, C., Martinez-Irizarry, M. M., Arvin, M. C., Orr, K. S., Manduchi, E., Kaestner, K. H., Brozinick, J. T., Linnemann, A. K.. 2025-02-20. IFN-α Induces Heterogenous ROS Production in Human β-Cells. https://doi.org/10.1101/2025.02.19.639120

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