Spermidine Dampens Inflammation by Directly Inhibiting Th17 Cytokine Production through a PRDX1 Associated Antioxidant Pathway

Abstract

The activation of IL-17 signaling has been linked to the pathogenesis of many chronic, inflammatory lung diseases including cystic fibrosis. Through unbiased single-cell RNAseq screening, we found that IL-17+ T cells highly express Srm and Smox, which encode two key enzymes involved in spermidine synthesis, spermidine synthase and spermine oxidase respectively. Spermidine has been shown to reduce inflammation by regulating macrophage activation and balancing Th17/Treg differentiation; however its direct effects on Th17 cytokine production have not been carefully investigated. Here, using already differentiated Th17 cells from cultured mouse splenocytes, we found that exogenous spermidine directly inhibits IL-1β/IL-23-induced IL-17 production. Blockade of endogenous spermidine synthesis enhanced IL-17 production above native levels, further supporting the notion that spermidine is a direct regulator of cytokine secretion independent of differentiation. In vivo, spermidine alleviates lung inflammation in both Pseudomonas aeruginosa (PA) and LPS induced acute lung injury models. Further RNA-seq analysis suggests that spermidine suppression of Th17 cytokine production is mediated through its PRDX1-dependent antioxidant activity. Our data suggests that spermidine is a direct regulator of Type-17 T cell cytokine production and has potent anti-inflammatory effects against lung inflammation.

Keywords

Spermidine / Interleukin-17 / Inflammation / Th17 Cells

Acknowledgements

This project is supported by HL137709, HL154231 and AI153549 from the National Institute of Health.

Author Contributions

Conceptualization, G.T.-N. and K.C.; Methodology, J.L., Y.J.K., S.C., X.A., L.F., C.E., D.L., A.A.F., Y.P., G.T.-N., K.C.; Validation, S.C. and L.F.; Formal Analysis, J.L., S.C. and D.L.; Investigation, X.X.; Resources, X.X.; Data Curation, X.X.; Writing—J.L., G.T.-N., K.C.; Writing—Review & Editing, J.L., Y.J.K., S.C., D.L., Y.P., G.T.-N., K.C.; Visualization, S.C., D.L.; Supervision, G.T.-N. and K.C.; Funding Acquisition, G.T.-N. and K.C.

Ethics Statement

All the mice were used with approval from the University of Pittsburgh Institutional Animal Care and Use Committee, Protocol #: 21028745, 2/2/2021.

Informed Consent Statement

Not applicable.

Data Availability Statement

R code used for RNA-seq analysis is available upon request. The sequencing data reported in this paper will be deposited in the Gene Expression Omnibus.

Funding

This project is supported by HL137709 (to K.C.), HL154231 (to G.T.-N.) and AI153549 (to G.T.-N.)

Declaration of Competing Interest

None of authors have any conflict of interest to report.

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