Uric acid priming in human monocytes is driven by the AKT-PRAS40 autophagy pathway.

Abstract

Metabolic triggers are important inducers of the inflammatory processes in gout. Whereas the high serum urate levels observed in patients with gout predispose them to the formation of monosodium urate (MSU) crystals, soluble urate also primes for inflammatory signals in cells responding to gout-related stimuli, but also in other common metabolic diseases. In this study, we investigated the mechanisms through which uric acid selectively lowers human blood monocyte production of the natural inhibitor IL-1 receptor antagonist (IL-1Ra) and shifts production toward the highly inflammatory IL-1β. Monocytes from healthy volunteers were first primed with uric acid for 24 h and then subjected to stimulation with lipopolysaccharide (LPS) in the presence or absence of MSU. Transcriptomic analysis revealed broad inflammatory pathways associated with uric acid priming, with NF-κB and mammalian target of rapamycin (mTOR) signaling strongly increased. Functional validation did not identify NF-κB or AMP-activated protein kinase phosphorylation, but uric acid priming induced phosphorylation of AKT and proline-rich AKT substrate 40 kDa (PRAS 40), which in turn activated mTOR. Subsequently, Western blot for the autophagic structure LC3-I and LC3-II (microtubule-associated protein 1A/1B-light chain 3) fractions, as well as fluorescence microscopy of LC3-GFP-overexpressing HeLa cells, revealed lower autophagic activity in cells exposed to uric acid compared with control conditions. Interestingly, reactive oxygen species production was diminished by uric acid priming. Thus, the Akt-PRAS40 pathway is activated by uric acid, which inhibits autophagy and recapitulates the uric acid-induced proinflammatory cytokine phenotype.

• Authors: Tania O Crişan, Maartje C P Cleophas, Boris Novakovic, Kathrin Erler, Frank L van de Veerdonk, Hendrik G Stunnenberg, Mihai G Netea, Charles A Dinarello, Leo A B Joosten
• Journal: Proceedings of the National Academy of Sciences of the United States of America (Proc Natl Acad Sci U S A) Vol. 114 Issue 21 Pg. 5485-5490 (05 23 2017) ISSN: 1091-6490 [Electronic] United States
• PMID: 28484006 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• AKT1S1 protein, human
• Adaptor Proteins, Signal Transducing
• IL1B protein, human
• Interleukin 1 Receptor Antagonist Protein
• Interleukin-1beta
• Reactive Oxygen Species
• Uric Acid
• Urate Oxidase
• Proto-Oncogene Proteins c-akt

Topics

• Adaptor Proteins, Signal Transducing (metabolism)
• Animals
• Autophagy
• Disease Models, Animal
• Gout (metabolism)
• Granulomatous Disease, Chronic (metabolism)
• HeLa Cells
• Humans
• Hyperuricemia (metabolism)
• Interleukin 1 Receptor Antagonist Protein (metabolism)
• Interleukin-1beta (metabolism)
• Mice
• Monocytes (metabolism)
• Proto-Oncogene Proteins c-akt (metabolism)
• Reactive Oxygen Species (metabolism)
• Transcriptome
• Urate Oxidase (antagonists & inhibitors)
• Uric Acid (metabolism)