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Nat Chem Biol. 2014 Dec;10(12):1043-8. doi: 10.1038/nchembio.1661. Epub 2014 Oct 26.

Hydrolysis of 2'3'-cGAMP by ENPP1 and design of nonhydrolyzable analogs.

Author information

1
Department of Systems Biology, Harvard Medical School, Boston, Massachusetts, USA.
2
Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, Massachusetts, USA.
3
Sanford Children's Health Research Center, Sanford-Burnham Medical Research Institute, La Jolla, California, USA.

Abstract

Agonists of mouse STING (TMEM173) shrink and even cure solid tumors by activating innate immunity; human STING (hSTING) agonists are needed to test this therapeutic hypothesis in humans. The endogenous STING agonist is 2'3'-cGAMP, a second messenger that signals the presence of cytosolic double-stranded DNA. We report activity-guided partial purification and identification of ecto-nucleotide pyrophosphatase/phosphodiesterase (ENPP1) to be the dominant 2'3'-cGAMP hydrolyzing activity in cultured cells. The hydrolysis activity of ENPP1 was confirmed using recombinant protein and was depleted in tissue extracts and plasma from Enpp1(-/-) mice. We synthesized a hydrolysis-resistant bisphosphothioate analog of 2'3'-cGAMP (2'3'-cG(s)A(s)MP) that has similar affinity for hSTING in vitro and is ten times more potent at inducing IFN-β secretion from human THP1 monocytes. Studies in mouse Enpp1(-/-) lung fibroblasts indicate that resistance to hydrolysis contributes substantially to its higher potency. 2'3'-cG(s)A(s)MP is therefore improved over natural 2'3'-cGAMP as a model agonist and has potential as a vaccine adjuvant and cancer therapeutic.

PMID:
25344812
PMCID:
PMC4232468
DOI:
10.1038/nchembio.1661
[Indexed for MEDLINE]
Free PMC Article

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