STING priming during immunotherapy
Author
Owczarska, Pamela Wiktoria
Term
4. term
Publication year
2020
Submitted on
2020-05-28
Pages
60
Abstract
Background: Immunotherapies may benefit from activating the STING pathway, which is triggered by cGAMP and promotes type I interferons and dendritic cell maturation. Cholesterol-targeting drugs can modulate this pathway. Aim: To assess whether nystatin or simvastatin can enhance (prime) cGAMP-mediated STING activation. Methods: Human monocyte-derived dendritic cells and murine bone marrow–derived dendritic cells (BALB/c and C57BL/6) were stimulated for 24 hours with nystatin or simvastatin with and without 2’3’-cGAMP. Maturation markers (CD86, MHC-II in mice; CD86, CD83, HLA-DR in humans) were quantified by flow cytometry. THP-1 cells (wild type and STING knockout) were differentiated into macrophages, stimulated similarly, and supernatants were assayed for type I interferon using a HEK-Blue IFN-α/β reporter. Results: In murine dendritic cells, nystatin combined with cGAMP significantly increased CD86 compared with nystatin alone, with no corresponding change in MHC-II. Simvastatin did not increase maturation markers in murine cells. In human dendritic cells, nystatin plus cGAMP tended to raise maturation compared with cGAMP alone, but without statistical significance; simvastatin data were inconclusive due to limited measurements. In THP-1 wild type, cGAMP induced type I interferon secretion, which was further increased by nystatin at both 50 µM and 5 µM cGAMP. Conclusion: Nystatin can enhance cGAMP-induced STING activation, reflected by increased dendritic cell maturation and type I interferon secretion, whereas simvastatin showed no clear effect. Further studies are needed to clarify nystatin’s mechanism and to validate these findings.
Baggrund: Immunterapier kan styrkes ved at aktivere STING-signalvejen, som udløses af cGAMP og fremmer type I interferoner og modning af dendritceller. Kolesterolmålrettede lægemidler kan påvirke denne vej. Formål: At undersøge om nystatin eller simvastatin kan forstærke (primere) cGAMP-medieret aktivering af STING. Metode: Humane monocyt-afledte dendritceller og murine knoglemarvs-afledte dendritceller (BALB/c og C57BL/6) blev stimuleret i 24 timer med nystatin eller simvastatin med og uden 2’3’-cGAMP. Modningsmarkører (CD86, MHC-II i mus; CD86, CD83, HLA-DR i menneske) blev målt ved flowcytometri. THP-1-celler (wild type og STING-knockout) blev differentieret til makrofager, stimuleret tilsvarende, og supernatant blev testet for type I interferon ved HEK-Blue IFN-α/β-reporter. Resultater: I murine dendritceller gav nystatin kombineret med cGAMP en signifikant stigning i CD86 sammenlignet med nystatin alene, uden tilsvarende ændring i MHC-II. Simvastatin øgede ikke modningsmarkører i murine celler. I humane dendritceller sås en tendens til højere modning med nystatin + cGAMP end med cGAMP alene, men uden statistisk signifikans; simvastatinresultaterne var uklare pga. begrænset datamængde. I THP-1 wild type udløste cGAMP sekretion af type I interferon, som yderligere blev øget ved tilsætning af nystatin ved både 50 µM og 5 µM cGAMP. Konklusion: Nystatin kan forstærke cGAMP-induceret STING-aktivering, afspejlet i øget dendritcellemodning og type I interferon-sekretion, mens simvastatin ikke viste en tydelig effekt. Yderligere studier er nødvendige for at klarlægge mekanismerne bag nystatins virkning og bekræfte fundene.
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Keywords