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Eradication of large established tumors in mice by combination immunotherapy that engages innate and adaptive immune responses

Abstract

Checkpoint blockade with antibodies specific for cytotoxic T lymphocyte–associated protein (CTLA)-4 or programmed cell death 1 (PDCD1; also known as PD-1) elicits durable tumor regression in metastatic cancer, but these dramatic responses are confined to a minority of patients. This suboptimal outcome is probably due in part to the complex network of immunosuppressive pathways present in advanced tumors, which are unlikely to be overcome by intervention at a single signaling checkpoint. Here we describe a combination immunotherapy that recruits a variety of innate and adaptive immune cells to eliminate large tumor burdens in syngeneic tumor models and a genetically engineered mouse model of melanoma; to our knowledge tumors of this size have not previously been curable by treatments relying on endogenous immunity. Maximal antitumor efficacy required four components: a tumor-antigen-targeting antibody, a recombinant interleukin-2 with an extended half-life, anti-PD-1 and a powerful T cell vaccine. Depletion experiments revealed that CD8+ T cells, cross-presenting dendritic cells and several other innate immune cell subsets were required for tumor regression. Effective treatment induced infiltration of immune cells and production of inflammatory cytokines in the tumor, enhanced antibody-mediated tumor antigen uptake and promoted antigen spreading. These results demonstrate the capacity of an elicited endogenous immune response to destroy large, established tumors and elucidate essential characteristics of combination immunotherapies that are capable of curing a majority of tumors in experimental settings typically viewed as intractable.

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Figure 1: AIPV immunotherapy cures large established tumors and establishes protective memory in multiple tumor models.
Figure 2: AIPV therapy primes sustained vaccine-specific T cell responses and remodels the microenvironment of established tumors.
Figure 3: AIPV therapy induces pronounced immune infiltration of tumors with efficacy dependent on innate and adaptive immune cells.
Figure 4: Combination therapy elicits antibody-enhanced antigen spreading and de novo T cell responses.
Figure 5: AIPV therapy induces de novo endogenous tumor-specific antibody responses.
Figure 6: AIPV with a trivalent vaccine is curative for established B16F10 tumors and induces regression in Braf/Pten autochthonous melanoma.

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Acknowledgements

This work was supported in part by the Koch Institute Support (core) grant P30-CA14051 from the National Cancer Institute, the US National Institutes of Health (NIH) grant CA174795 (K.D.W.), the Bridge Project partnership between the Koch Institute for Integrative Cancer Research and the Dana Farber–Harvard Cancer Center (DF–HCC) (D.J.I.), the V Foundation (D.J.I.) and the Ragon Institute (D.J.I.). K.D.M. and J.M.E. are supported by the Fannie and John Hertz Foundation Fellowship; K.D.M., C.F.O., J.M.E., B.H.K. and E.F.Z. are supported by NSF Graduate Research Fellowships; A.M.R. is supported by the NIGMS–NIH Interdepartmental Biotechnology Training Program (NIH #T32GM008334); A.T. is supported by the Siebel Scholarship; and G.L.S. was supported by the NIH with a Ruth L. Kirschstein National Research Service Award (CA180586). We thank T.C. Wu (Johns Hopkins University) for kindly providing the TC-1 tumor cells, D. Sabatini (Whitehead Institute) for providing the Cas9 constructs and G. Dranoff (Dana-Farber Cancer Institute) for providing the DD-Her2/neu and B16-OVA cells. We thank M. Ghebremichael (Ragon Institute of MGH, MIT and Harvard) for helpful statistical advice. We thank the Koch Institute Swanson Biotechnology Center for technical support, specifically the applied therapeutics and whole-animal imaging core facility, the histology and the flow cytometry core facilities. D.J.I. is an investigator of the Howard Hughes Medical Institute.

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Contributions

K.D.M., C.F.O., D.J.I. and K.D.W. designed the studies and wrote the manuscript; K.D.M. and C.F.O. carried out experiments; E.F.Z., A.T., B.H.K., M.J.K. and H.S. assisted with protein production; J.M.E. assisted in generating the Trp2-knockout line using CRISPR–Cas9; K.R., A.T., E.F.Z., W.A., R.T.W., A.M.R., R.L.K., N.K.M. and K.H. assisted with experiments; G.L.S. designed and analyzed intratumoral Luminex, and assisted in flow cytometry design and writing of the manuscript; D.A.L. aided with multivariate analysis of intratumoral Luminex data; G.L.S. and M.H.Z. conducted intratumoral Luminex and assisted with Braf/Pten mouse experiments; S.K. assisted with immunofluorescence microscopy; and J.R.C. supplied 2.5F–Fc reagents.

Corresponding authors

Correspondence to K Dane Wittrup or Darrell J Irvine.

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Competing interests

D.J.I. holds equity in Vedantra Pharmaceuticals, which holds a license to the amphiphile–vaccine technology used in these studies.

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Moynihan, K., Opel, C., Szeto, G. et al. Eradication of large established tumors in mice by combination immunotherapy that engages innate and adaptive immune responses. Nat Med 22, 1402–1410 (2016). https://doi.org/10.1038/nm.4200

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