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Selective blockade of the lyso-PS lipase ABHD12 stimulates immune responses in vivo

Abstract

ABHD12 metabolizes bioactive lysophospholipids, including lysophosphatidylserine (lyso-PS). Deleterious mutations in human ABHD12 cause the neurological disease PHARC, and ABHD12–/– mice display PHARC-like phenotypes, including hearing loss, along with elevated brain lyso-PS and features of stimulated innate immune cell function. Here, we develop a selective and in vivo–active inhibitor of ABHD12 termed DO264 and show that this compound elevates lyso-PS in mouse brain and primary human macrophages. Unlike ABHD12–/– mice, adult mice treated with DO264 exhibited minimal perturbations in auditory function. On the other hand, both DO264-treated and ABHD12–/– mice displayed heightened immunological responses to lymphocytic choriomeningitis virus (LCMV) clone 13 infection that manifested as severe lung pathology with elevated proinflammatory chemokines. These results reveal similarities and differences in the phenotypic impact of pharmacological versus genetic blockade of ABHD12 and point to a key role for this enzyme in regulating immunostimulatory lipid pathways in vivo.

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Fig. 1: Development of a high-throughput screening (HTS) assay for ABHD12 inhibitors.
Fig. 2: Discovery and optimization of (thio)urea inhibitors of ABHD12.
Fig. 3: DO264 inhibits ABHD12 and increases lyso-PS content in human monocytic cells.
Fig. 4: DO264 inhibits ABHD12 and increases brain lyso-PS content in vivo.
Fig. 5: Effects of ABHD12 inhibition on mouse auditory capacity and brain lyso-PS content in vivo.
Fig. 6: Heightened immunopathological responses to LCMV-clone 13 (Cl13) infection of mice with genetic or pharmacological inactivation of ABHD12.

Data availability

All data generated or analyzed during this study are included in this published article (and its supplementary information files) or are available from the corresponding author on reasonable request.

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Acknowledgements

This work was supported by the NIH (DA033760, NS092980, AI123210, AI117175) and the Skaggs Institute for Chemical Biology. J.B. is supported by a fellowship from Celgene. We thank J. Olucha for recombinant expression of mABHD12 in HEK293F cells; J. Chen (Automated Synthesis Facility at Scripps Research) for measuring enantiopurity of (S)-DO271; C.E. Moore and M. Gembicky (UCSD) for X-ray crystallographic analysis of DO253; and J. Wang and C. Chen (Pharmaron) for the measurement of tissue drug concentration.

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D.O., T.-A.I., V.F.V., J.R.T., and B.F.C. conceived the project and designed the experiments. T.-A.I. performed the HTS study. S.B. assisted with HTS assay development, and H.R. provided the small molecule library and screening instruments. D.O. synthesized and chemically characterized the compounds. J.J.H. synthesized JJH350. D.O., T.-A.I., A.R., O.U., and H.J. performed the biochemical and cellular experiments. J.B. performed the experiments with primary human macrophages. D.O. and T.-A.I. performed the in vivo compound dosing and lipidomic studies. A.R. performed the auditory tests. V.F.V. and J.R.T. performed the immunological experiments. T.-A.I., D.O., J.R.T. and B.F.C. wrote the manuscript.

Corresponding authors

Correspondence to John R. Teijaro or Benjamin F. Cravatt.

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O.U. and A.R. are employees of and B.F.C. is a founder and advisor to Abide Therapeutics, a biotechnology company interested in developing serine hydrolase inhibitors and therapeutics.

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Supplementary information

Supplementary Information

Supplementary Tables 1–2, Supplementary Figures 1–29

Reporting Summary

Supplementary Note

Synthetic Procedures

Supplementary Table 3

Complete mass spectrometry–based ABPP data

Supplementary Table 4

Complete targeted lipidomics data

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Ogasawara, D., Ichu, TA., Vartabedian, V.F. et al. Selective blockade of the lyso-PS lipase ABHD12 stimulates immune responses in vivo. Nat Chem Biol 14, 1099–1108 (2018). https://doi.org/10.1038/s41589-018-0155-8

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