(280) Peroxynitrite Biochemical Signaling Regulates Micro RNA (miR21) Activation of NLRP3 Inflammasome and is Key to PCOS Severity

Abstract Authors: Annika Chatterjee¹, Ayushi Trivedi², Shuo Xiao³, Stephen C. Bondy²
1. University High School, Irvine, CA, USA
2. Environmental and Occupational Health, University of California, Irvine, Irvine, CA USA
3. Department of Pharmacology and Toxicology, Ernest Mario School of Pharmacy, Environmental and Occupational Health Sciences Institute (EOHSI), Rutgers University,  Piscataway, NJ USA

Abstract Text:

Biochemical signaling plays a key role in molecular pathways in various disease processes of women’s reproductive health. Though the role of oxidative stress in various reproductive pathology is known, precise role of peroxynitrite (ONOO-) a key nitric oxide metabolite in stimulating micro RNAs in connecting molecular biology of polycystic ovarian syndrome is unknown. I hypothesize that peroxynitrite will trigger the activation of micro-RNA (miR)-21 that ultimately will activate inflammasome in mouse organoids and human granulosa (KGN) cells. Using both a novel mouse ovarian organoid system and an in vitro cellular model, I show that both systems produced a significant increase in miR21 and a parallel increase in ONOO-. Specific inhibitors of peroxynitrite such as phenyl boronic acid, DMPO and apocynin, a NOX2 inhibitor significantly decreased miR21 levels suggesting that ONOO- is involved in miR21 activation. It also showed that ONOO- resulted from an activated NOX2 enzyme since apocynin, an inhibitor of p47 phox translocation to the membrane blocked ONOO- formation. To study whether ONOO- also modulated the downstream inflammasome activation, I conducted dual labelled immunofluorescence microscopy for visualizing NLRP3-ASC2 colocalization, Results showed that ONOO- inhibitors significantly decreased inflammasome activation and release of IL1β, a secreted proinflammatory cytokine. In conclusion, my data represented a unique redox pathway that was upstream of miR21-NLRP3 axis and may be used as a therapeutic target in PCOS.