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A natural killer–dendritic cell axis defines checkpoint therapy–responsive tumor microenvironments

Nature Medicine volume 24, pages 1178–1191 (2018)Cite this article

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Abstract

Intratumoral stimulatory dendritic cells (SDCs) play an important role in stimulating cytotoxic T cells and driving immune responses against cancer. Understanding the mechanisms that regulate their abundance in the tumor microenvironment (TME) could unveil new therapeutic opportunities. We find that in human melanoma, SDC abundance is associated with intratumoral expression of the gene encoding the cytokine FLT3LG. FLT3LG is predominantly produced by lymphocytes, notably natural killer (NK) cells in mouse and human tumors. NK cells stably form conjugates with SDCs in the mouse TME, and genetic and cellular ablation of NK cells in mice demonstrates their importance in positively regulating SDC abundance in tumor through production of FLT3L. Although anti-PD-1 ‘checkpoint’ immunotherapy for cancer largely targets T cells, we find that NK cell frequency correlates with protective SDCs in human cancers, with patient responsiveness to anti-PD-1 immunotherapy, and with increased overall survival. Our studies reveal that innate immune SDCs and NK cells cluster together as an excellent prognostic tool for T cell–directed immunotherapy and that these innate cells are necessary for enhanced T cell tumor responses, suggesting this axis as a target for new therapies.

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Fig. 1: BDCA-3+ DCs define overall outcome in melanoma patients and predict responsiveness to anti-PD-1 immunotherapy.
Fig. 2: Tumor-resident lymphocytes produce FLT3L.
Fig. 3: FLT3L production by NK cells controls the levels of CD103+ SDCs in tumor.
Fig. 4: NK cells make frequent, stable interactions with XCR1+ DCs and provide prosurvival signals.
Fig. 5: BDCA-3+ DC levels correlate with levels of NK cells in the human melanoma TME.
Fig. 6: NK cell and BDCA-3+ DC levels uniquely correlate with anti-PD-1 responsiveness in patients with melanoma.

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Acknowledgements

We thank L. Lanier, J. Roose and L. Fong for advice, and we thank M. Spasic and N. Khurana for support with response data. This work was supported by National Institutes of Health (NIH) grant R01CA197363, awarded to M.F.K. Acquisition and processing of human melanoma samples in cohort A described in this study was funded in part by contributions from AbbVie, Amgen, and Bristol-Myers Squibb as members of the Immunoprofiler Consortium. Further support came from NIH grant 5P30CA082103, awarded to the University of California, San Francisco (UCSF) Hellen Diller Family Comprehensive Cancer Center. M.B. was supported by the Genentech Predoctoral Research Fellowship, the Margaret A. Cunningham Immune Mechanisms in Cancer Research Fellowship Award, and the Achievement Reward for College Scientists Scholarship. K.C.B. was supported by a postdoctoral fellowship from the Cancer Research Institute and Fibrolamellar Cancer Foundation.

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Authors and Affiliations

  1. Department of Pathology, University of California San Francisco, San Francisco, CA, USA

    Kevin C. Barry, Joy Hsu, Miranda L. Broz, Francisco J. Cueto, Mikhail Binnewies, Alexis J. Combes, Amanda E. Nelson, Raj Kumar, Bushra Samad, Vincent Chan & Matthew F. Krummel

  2. UCSF Immunoprofiler Initiative, University of California San Francisco, San Francisco, CA, USA

    Kevin C. Barry, Joy Hsu, Miranda L. Broz, Alexis J. Combes, Amanda E. Nelson, Kimberly Loo, Raj Kumar, Bushra Samad, Saurabh Asthana, Vincent Chan & Matthew F. Krummel

  3. Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC), Madrid, Spain

    Francisco J. Cueto

  4. Department of Biochemistry, Faculty of Medicine, Universidad Autónoma de Madrid, Madrid, Spain

    Francisco J. Cueto

  5. Melanoma Clinical Research Unit, University of California San Francisco, San Francisco, CA, USA

    Kimberly Loo

  6. Department of Dermatology, University of California San Francisco, San Francisco, CA, USA

    Kimberly Loo, Michael D. Rosenblum, Michael D. Alvarado & Adil I. Daud

  7. Department of Laboratory Medicine, University of California San Francisco, San Francisco, CA, USA

    Denise M. Wolf

  8. Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY, USA

    Dusan Bogunovic

  9. Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA

    Nina Bhardwaj

  10. Department of Otolaryngology, University of California San Francisco, San Francisco, CA, USA

    Patrick K. Ha & William R. Ryan

  11. Pionyr Immunotherapeutics, San Francisco, CA, USA

    Joshua L. Pollack

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Contributions

K.C.B. designed and performed the experiments and wrote and edited the manuscript. J.H. assisted in analysis of tumor-infiltrating myeloid populations and data analysis. M.L.B. designed and performed experiments with melanoma cohort B. F.J.C. assisted in imaging data analysis. M.B. assisted in analysis of tumor-infiltrating myeloid populations and T cell–depletion experiments. A.J.C. assisted in analysis of HNSCC tumor samples. R.K. and A.E.N. assisted in analysis of tumor-infiltrating myeloid populations. K.L. and A.I.D. provided human melanoma biopsies, clinical data, and edited the manuscript. M.D.R. read the manuscript and provided useful discussion. M.D.A. provided human melanoma biopsies and clinical data. D.B. and N.B. provided metastatic melanoma data, and D.B. edited the manuscript. D.M.W. performed statistical analyses. P.K.H and W.R.R provided human HNSCC samples. J.L.P., B.S., and S.A. provided bioinformatics analyses. V.C. managed sample collection, assisted in analysis of tumor-infiltrating myeloid populations, read the manuscript, and provided useful discussion. M.F.K. conceived the project and wrote and edited the manuscript.

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Correspondence to Matthew F. Krummel.

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J.L.P. was an employee at Pionyr Immunotherapeutics at the time of manuscript writing. The other authors declare no competing interests.

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Supplementary Text and Figures

Supplementary Figures 1–7 and Supplementary Tables 1–3

Reporting Summary

Supplementary Video 1

Stable interaction of NK cells and XCR1+ DCs

Supplementary Video 2

NK cells distant from XCR1+ DCs have increased motility

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Barry, K.C., Hsu, J., Broz, M.L. et al. A natural killer–dendritic cell axis defines checkpoint therapy–responsive tumor microenvironments. Nat Med 24, 1178–1191 (2018). https://doi.org/10.1038/s41591-018-0085-8

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