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Reversal of indoleamine 2,3-dioxygenase–mediated cancer immune suppression by systemic kynurenine depletion with a therapeutic enzyme

Abstract

Increased tryptophan (Trp) catabolism in the tumor microenvironment (TME) can mediate immune suppression by upregulation of interferon (IFN)-γ-inducible indoleamine 2,3-dioxygenase (IDO1) and/or ectopic expression of the predominantly liver-restricted enzyme tryptophan 2,3-dioxygenase (TDO)1,2,3,4,5. Whether these effects are due to Trp depletion in the TME or mediated by the accumulation of the IDO1 and/or TDO (hereafter referred to as IDO1/TDO) product kynurenine (Kyn) remains controversial5,6,7,8,9,10,11,12,13. Here we show that administration of a pharmacologically optimized enzyme (PEGylated kynureninase; hereafter referred to as PEG-KYNase) that degrades Kyn into immunologically inert, nontoxic and readily cleared metabolites inhibits tumor growth. Enzyme treatment was associated with a marked increase in the tumor infiltration and proliferation of polyfunctional CD8+ lymphocytes. We show that PEG-KYNase administration had substantial therapeutic effects when combined with approved checkpoint inhibitors or with a cancer vaccine for the treatment of large B16-F10 melanoma, 4T1 breast carcinoma or CT26 colon carcinoma tumors. PEG-KYNase mediated prolonged depletion of Kyn in the TME and reversed the modulatory effects of IDO1/TDO upregulation in the TME.

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Figure 1: Kynureninase administration reduces tumor growth by depleting kynurenine from the tumor microenvironment.
Figure 2: Kynureninase treatment increases the frequency of effector CD8+ TILs.
Figure 3: Combination of PEG-kynureninase with other immunotherapies for the treatment of large, established tumors in multiple tumor models.

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Acknowledgements

We are grateful to N. Ashoura for assistance with various aspects of this work, data interpretation and comments on the manuscript. This work was supported by funding from NIH 1 RO1 CA189623 (E. Stone and G.G.), the Cancer Prevention and Research Institute of Texas grant DP150061 (E. Stone, L.I.R.E. and G.G.) and Kyn Therapeutics (G.G. and E. Stone). Support from the American Cancer Society was provided to T.A.T. (Postdoctoral Fellowship 126584-PF-14-216-01-TBF), N.M. (Postdoctoral Fellowship 123506-PF-13-354-01-CDD) and J.B. (Postdoctoral Fellowship 128252-PF-15-143-01-CDD). M.D. acknowledges support by a postdoctoral fellowship from the Cancer Prevention and Research Institute of Texas (RP140108).

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Contributions

T.A.T., K.C.G., N.M. and M.D. designed and performed key experiments; J.B., C.L., A.Q., B.T., Y.T., W.-C.L., C.S.K., K.F. and M.S.Y. expressed, cloned, characterized and prepared enzymes for in vitro and in vivo studies; B.T. and Y.K. developed and performed pharmacokinetic assays; J.D.D., M.M., X.M.Z., G.F., K.M., S.C. and T.H.S. designed and performed certain in vivo and in vitro experiments; E. Sentandreu and S.T. performed metabolomics analyses; T.A.T., K.C.G., M.D., J.D.D., L.I.R.E., G.G. and E. Stone interpreted the data; and G.G., T.A.T., K.C.G. and E. Stone wrote the manuscript.

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Correspondence to Everett Stone or George Georgiou.

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Competing interests

G.G. and E. Stone are inventors on intellectual property related to this work, and G.G., E. Stone, X.M.Z., K.M., S.C. and M.M. have equity interest in Kyn Therapeutics, a company pursuing the commercial development of this technology.

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Triplett, T., Garrison, K., Marshall, N. et al. Reversal of indoleamine 2,3-dioxygenase–mediated cancer immune suppression by systemic kynurenine depletion with a therapeutic enzyme. Nat Biotechnol 36, 758–764 (2018). https://doi.org/10.1038/nbt.4180

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