1810-P: Beta-Cell Extracellular Vesicles—Transfer to and Impact on Neighboring Cells

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Introduction and Objective: Extracellular vesicles (EVs) are small membrane-bound particles secreted by cells, playing a crucial role in intercellular communication and cellular homeostasis.Methods: Human EndoC βH1 beta cells were obtained from Human Cell Design. To assess impacts of EV transfer on recipient cells, small EVs from cells treated with or without a cytokine mix were isolated using ultracentrifugation and wild-type recipient EndoC βH1 cells were treated with 5×108 EVs from each group for 48 hours, followed by total and small RNA-sequencing. Data analysis was performed using Qiagen’s Ingenuity Pathway Analysis (IPA). To fluorescently label EVs for quantification of transfer, EndoC βH1-CD81-mCherry and EndoC βH1-iRFP720 cells were generated by transfecting plasmids encoding CD81-mCherry and iRFP720 into EndoC βH1 cells. To assess impacts of proinflammatory cytokine treatment on EV transfer, cells were co-seeded on the same plate and after 48 hours of treatment with or without 5 ng/mL IL-1β, 100 ng/mL IFN-γ, CD81+ EV transfer to neighboring cells was assessed using flow cytometry quantification of mCherry+ iRFP720+ cells.Results: Treatment with EVs from cytokine-treated parent cells, yielded differential gene expression in recipient cells, marked by increased type I and type II interferon signaling, antigen presentation, macrophage classical activation signaling, and class I MHC-mediated antigen processing. Treatment of cocultured cells with cytokine mix significantly enhanced EV transfer, as evidenced by a 1.7-fold increase in mCherry+ iRFP720+ cells compared to controls (p<0.05).Conclusion: Inflammatory EVs exhibited altered mRNA and miRNA expression profiles, consistent with activation of interferon signaling and antigen presentation in recipient cells. Further, cytokine-exposure enhanced EV transfer to neighboring cells. These findings suggest that beta cell derived EVs may play a role in propagating and amplifying beta cell cytokine signaling.

Disclosure

J. Xu: None. E.K. Sims: Consultant; Sanofi. Speaker’s Bureau; Med Learning Group. Other Relationship; American Diabetes Association.



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