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DPP8 and DPP9 inhibition induces pro-caspase-1-dependent monocyte and macrophage pyroptosis

Nature Chemical Biology volume 13, pages 46–53 (2017)Cite this article

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Abstract

Val-boroPro (Talabostat, PT-100), a nonselective inhibitor of post-proline cleaving serine proteases, stimulates mammalian immune systems through an unknown mechanism of action. Despite this lack of mechanistic understanding, Val-boroPro has attracted substantial interest as a potential anticancer agent, reaching phase 3 trials in humans. Here we show that Val-boroPro stimulates the immune system by triggering a proinflammatory form of cell death in monocytes and macrophages known as pyroptosis. We demonstrate that the inhibition of two serine proteases, DPP8 and DPP9, activates the pro-protein form of caspase-1 independent of the inflammasome adaptor ASC. Activated pro-caspase-1 does not efficiently process itself or IL-1β but does cleave and activate gasdermin D to induce pyroptosis. Mice lacking caspase-1 do not show immune stimulation after treatment with Val-boroPro. Our data identify what is to our knowledge the first small molecule that induces pyroptosis and reveals a new checkpoint that controls the activation of the innate immune system.

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Figure 1: Val-boroPro is cytotoxic to monocytes and macrophages.
Figure 2: DPP8/9 inhibition induces cell death.
Figure 3: DPP8/9 inhibitor cytotoxicity is caspase-1 dependent.
Figure 4: DPP8/9 inhibition activates pro-caspase-1 without autoproteolysis.
Figure 5: GSDMD is cleaved after DPP8/9 inhibition and contributes to cell death.
Figure 6: Val-boroPro does not induce cytokines in caspase-1 knockout mice.

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Acknowledgements

We thank S. Fujisawa for microscopy assistance, E. De Stanchina and B. Qeriqi for assistance harvesting mBMDMs, C. Taabazuing for helpful comments and A. Kentsis and F. Brown (Molecular Pharmacology Program, Memorial Sloan Kettering Cancer Center) for the HL-60 cell line. This work was supported by the Josie Robertson Foundation (D.A.B.), the MSKCC Core Grant (P30 CA008748), the NCI (grant no. U54CA112962 to T.R.G.), HHMI (T.R.G.) and the NIH (CA174008-01A1 to W.W.B. and NIH NIGMS T32 GM115327-Tan to D.C.J.).

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Authors and Affiliations

  1. Chemical Biology Program, Memorial Sloan Kettering Cancer Center, New York, New York, USA

    Marian C Okondo & Daniel A Bachovchin

  2. Tri-Institutional PhD Program in Chemical Biology, Memorial Sloan Kettering Cancer Center, New York, New York, USA

    Darren C Johnson, Eun Bin Go, Ashley J Chui & Daniel A Bachovchin

  3. Graduate Program in Pharmacology, Weill Cornell Graduate School of Medical Sciences, New York, New York, USA

    Ramya Sridharan, Mitchell S Wang & Daniel A Bachovchin

  4. Department of Developmental, Chemical & Molecular Biology, Sackler School of Graduate Biomedical Sciences, Tufts University, Boston, Massachusetts, USA

    Sarah E Poplawski, Wengen Wu, Yuxin Liu, Jack H Lai, David G Sanford, Michael O Arciprete & William W Bachovchin

  5. The Eli and Edythe L. Broad Institute, Cambridge, Massachusetts, USA

    Todd R Golub

  6. Department of Pediatric Oncology, Dana-Farber Cancer Institute, Boston, Massachusetts, USA

    Todd R Golub

  7. Harvard Medical School, Boston, Massachusetts, USA

    Todd R Golub

  8. Howard Hughes Medical Institute, Chevy Chase, Maryland, USA

    Todd R Golub

  9. Arisaph Pharmaceuticals, Boston, Massachusetts, USA

    William W Bachovchin

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Contributions

D.A.B. conceived and directed the project, performed experiments, analyzed data and wrote the paper; M.C.O., D.C.J., R.S., E.B.G., A.J.C. and M.S.W. performed experiments and analyzed data; S.E.P. performed the in vivo mouse experiments and the intracellular DPP8/9 inhibition experiment; W.W.B. and D.G.S. directed the in vivo mouse experiments; W.W., Y.L. and J.H.L. synthesized Val-boroPro, 1G244, L-allo-isoleucin-isoindoline, L-allo-isoleucine-thiazolidine, compound 5385 and FP-biotin; M.O.A. characterized compound 5385. T.R.G. and W.W.B. helped plan the study.

Corresponding author

Correspondence to Daniel A Bachovchin.

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Competing interests

W.W.B. is a co-founder, advisor and board member of Arisaph Pharmaceuticals, a biotechnology company interested in developing boronic acid–based inhibitors of serine proteases as therapeutics.

Supplementary information

Supplementary Text and Figures

Supplementary Results, Supplementary Tables 1 and 2 and Supplementary Figures 1–12. (PDF 14969 kb)

Supplementary Video 1

DMSO-treated RAW 264.7 cells. (MOV 17579 kb)

Supplementary Video 2

Etoposide-induced apoptosis in RAW 264.7 cells. (MOV 13930 kb)

Supplementary Video 3

Val-boroPro-induced pyroptosis in RAW 264.7 cells. (MOV 15726 kb)

Supplementary Video 4

1G244-induced pyroptosis in RAW 264.7 cells. (MOV 13860 kb)

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Okondo, M., Johnson, D., Sridharan, R. et al. DPP8 and DPP9 inhibition induces pro-caspase-1-dependent monocyte and macrophage pyroptosis. Nat Chem Biol 13, 46–53 (2017). https://doi.org/10.1038/nchembio.2229

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