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Diabetes reversal by inhibition of the low-molecular-weight tyrosine phosphatase

Abstract

Obesity-associated insulin resistance plays a central role in type 2 diabetes. As such, tyrosine phosphatases that dephosphorylate the insulin receptor (IR) are potential therapeutic targets. The low-molecular-weight protein tyrosine phosphatase (LMPTP) is a proposed IR phosphatase, yet its role in insulin signaling in vivo has not been defined. Here we show that global and liver-specific LMPTP deletion protects mice from high-fat diet-induced diabetes without affecting body weight. To examine the role of the catalytic activity of LMPTP, we developed a small-molecule inhibitor with a novel uncompetitive mechanism, a unique binding site at the opening of the catalytic pocket, and an exquisite selectivity over other phosphatases. This inhibitor is orally bioavailable, and it increases liver IR phosphorylation in vivo and reverses high-fat diet-induced diabetes. Our findings suggest that LMPTP is a key promoter of insulin resistance and that LMPTP inhibitors would be beneficial for treating type 2 diabetes.

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Figure 1: Genetic deletion of LMPTP improves glucose tolerance of obese mice and increases liver insulin receptor signaling.
Figure 2: High-throughput screen of NIH chemical library identifies selective inhibitors of LMPTP.
Figure 3: The LMPTP inhibitor series is selective and displays an uncompetitive mechanism of action.
Figure 4: Structural determinants of LMPTP inhibition.
Figure 5: Compound 23 increases liver cell insulin signaling, is orally bioavailable and reverses diabetes in obese mice.

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Acknowledgements

The authors are grateful to L. Tautz at the Sanford Burnham Prebys Medical Discovery Institute for providing recombinant VHR and LYP proteins, to E. Santelli for critical review of the manuscript, to the University of California Davis Mouse Biology Program for help with image preparation, to S. Gupta for assistance with sample irradiation at the Advanced Light Source of Lawrence Berkeley National Laboratory, and to Z. Mikulski and A. Lamberth for help with histological analysis at the Histology and Microscopy Core at the La Jolla Institute for Allergy and Immunology. This work was supported by grants R03DA033986 (to N.B.) and R01DK106233 from the National Institutes of Health (to N.B. and A.B.P.). S.M.S. was supported by the American Diabetes Association Pathway to Stop Diabetes Grant 1-15-INI-13 and by the University of California San Diego Diabetes Research Center grant P30DK063491 from the National Institutes of Health. The X-ray footprinting was carried out by S. Gupta at beamline 3.2.1 of the Advanced Light Source of Lawrence Berkeley National Laboratory, which is supported by the Director, Office of Science, Office of Basic Energy Sciences, of the US Department of Energy under Contract No. DE-AC02-05CH11231. This is manuscript #1882 from the La Jolla Institute for Allergy and Immunology.

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S.M.S., A.E.A., T.D.Y.C., T.M., S.S., L.A.B., R.C.L., A.B.P. and N.B. participated in study conception and design. S.M.S., V.Z., M.R.B., Y.L., A.B. and S.S. performed in vivo experiments. S.M.S., V.Z. and M.R.B. performed cell biology experiments. S.M.S., V.Z., M.P.H., M.R.B., F.Y. and S.K. performed in vitro enzymatic assays. R.J.A., J.Z., S.R.G. and A.B.P. designed chemical compounds and/or performed chemical syntheses. R.J.A. and A.B.P. coordinated compound stability and PK studies. A.E.A., M.P.H., M.R.B., A.A.B., Y.L., G.W.C. and L.A.B. produced recombinant proteins. A.A.B. performed ITC experiments. A.E.A., G.W.C., J.Y. and L.A.B. performed NMR and X-ray crystallography experiments. J.K. performed hydroxyl radical footprinting experiments. S.M.S., A.E.A., V.Z., R.J.A., M.P.H., J.Z., S.R.G., M.R.B., F.Y., A.A.B., J.K., Y.L., G.W.C., S.K., J.Y., A.B., T.D.Y.C., T.M., S.S., L.A.B., R.C.L., A.B.P. and N.B. analyzed and interpreted data. S.M.S., R.C.L., A.B.P. and N.B. wrote the manuscript.

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Correspondence to Nunzio Bottini.

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The La Jolla Institute for Allergy and Immunology and Sanford Burnham Medical Discovery Institute hold a pending patent, WO 2016/061280 A1, “Inhibitors of low molecular weight protein tyrosine phosphatase and uses thereof,” with N.B., J.Z., S.R.G., S.M.S., A.B.P., T.D.Y.C., M.P.H. and R.J.A. named as inventors.

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Stanford, S., Aleshin, A., Zhang, V. et al. Diabetes reversal by inhibition of the low-molecular-weight tyrosine phosphatase. Nat Chem Biol 13, 624–632 (2017). https://doi.org/10.1038/nchembio.2344

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