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PI3Kγδ inhibitor plus radiation enhances the antitumour immune effect of PD-1 blockade in syngenic murine breast cancer and humanised patient-derived xenograft model

  • Author Footnotes
    1 Contributed equally.
    Min Guk Han
    Footnotes
    1 Contributed equally.
    Affiliations
    Department of Tumour Biology, Graduate School of Medicine, Seoul National University, Seoul, South Korea

    Medical Science Research Institute, Seoul National University Bundang Hospital, Seongnam, Seoul, South Korea
    Search for articles by this author
  • Author Footnotes
    1 Contributed equally.
    Bum-Sup Jang
    Footnotes
    1 Contributed equally.
    Affiliations
    Department of Radiation Oncology, Seoul National University Bundang Hospital, Seongnam, Seoul, South Korea
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  • Mi Hyun Kang
    Affiliations
    Medical Science Research Institute, Seoul National University Bundang Hospital, Seongnam, Seoul, South Korea
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  • Deukchae Na
    Affiliations
    Institute of Convergence Medicine, Ewha Woman's University Mokdong Hospital, Seoul, South Korea
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  • In Ah Kim
    Correspondence
    Corresponding author: Department of Radiation Oncology, Seoul National University, College of Medicine, 173 Gumiro, Bundanggu, Seongnamsi, Kyeonggido, 13620, South Korea. Fax: +82(31) 787-4019.
    Affiliations
    Department of Tumour Biology, Graduate School of Medicine, Seoul National University, Seoul, South Korea

    Medical Science Research Institute, Seoul National University Bundang Hospital, Seongnam, Seoul, South Korea

    Department of Radiation Oncology, Seoul National University Bundang Hospital, Seongnam, Seoul, South Korea

    Department of Radiation Oncology and Cancer Research Institute, Seoul National University, College of Medicine, Seoul, South Korea
    Search for articles by this author
  • Author Footnotes
    1 Contributed equally.
Published:September 30, 2021DOI:https://doi.org/10.1016/j.ejca.2021.08.029

      Highlights

      • Combining PD-1 blockade, radiation therapy and phosphoinositide 3-kinaseγδ inhibitor were tested in breast cancer models.
      • Syngenic murine tumour and humanised patient-derived xenograft model were used.
      • The greatest anti-tumour effect was achieved with triple combination therapy.
      • Triple combination increased cytotoxic T-cells and decreased immune-suppressive cells.
      • This strategy could be a viable approach to overcome the therapeutic resistance of PD1 blockade.

      Abstract

      Introduction

      We hypothesised that the combined use of radiation therapy and a phosphoinositide 3-kinaseγδ inhibitor to reduce immune suppression would enhance the efficacy of an immune checkpoint inhibitor.

      Methods

      Murine breast cancer cells (4T1) were grown in both immune-competent and -deficient BALB/c mice, and tumours were irradiated by 3 fractions of 24 Gy. A PD-1 blockade and a phosphoinositide 3-kinase (PI3K)γδ inhibitor were then administered every other day for 2 weeks. The same experiments were performed in humanised patient-derived breast cancer xenograft model and its tumour was sequenced to identify immune-related pathways and profile infiltrated immune cells. Transcriptomic and clinical data were acquired from The Cancer Genome Atlas pan-cancer cohort, and the deconvolution algorithm was used to profile immune cell repertoire.

      Results

      Using a PI3Kγδ inhibitor, radiation therapy (RT) and PD-1 blockade significantly delayed primary tumour growth, boosted the abscopal effect and improved animal survival. RT significantly increased CD8+cytotoxic T-cell fractions, immune-suppressive regulatory T cells (Tregs), myeloid-derived suppressor cells and M2 tumour-associated macrophages (TAMs). However, the PI3Kγδ inhibitor significantly lowered the proportions of Tregs, myeloid-derived suppressor cells and M2 TAMs, achieving dramatic gains in splenic, nodal, and tumour CD8+ T-cell populations after triple combination therapy. In a humanised patient-derived breast cancer xenograft model, triple combination therapy significantly delayed tumour growth and decreased immune-suppressive pathways. In The Cancer Genome Atlas cohort, high Treg/CD8+ T cell and M2/M1 TAM ratios were associated with poor overall patient survival.

      Conclusion

      These findings indicate PI3Kγ and PI3Kδ are clinically relevant targets in an immunosuppressive TME, and combining PI3Kγδ inhibitor, RT and PD-1 blockade may overcome the therapeutic resistance of immunologically cold tumours.

      Synopsis

      Combining PI3Kγδ inhibitor, RT, and PD-1 blockade may be a viable clinical approach, helping to overcome the therapeutic resistance of immunologically cold tumours such as breast cancer.

      Keywords

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      References

        • Robert C.
        • Ribas A.
        • Hamid O.
        • Daud A.
        • Wolchok J.D.
        • Joshua A.M.
        • et al.
        Durable complete response after discontinuation of pembrolizumab in patients with metastatic melanoma.
        J Clin Oncol. 2018; 36: 1668-1674
        • Socinski M.A.
        • Jotte R.M.
        • Cappuzzo F.
        • Orlandi F.
        • Stroyakovskiy D.
        • Nogami N.
        • et al.
        Atezolizumab for first-line treatment of metastatic nonsquamous NSCLC.
        N Engl J Med. 2018; 378: 2288-2301
        • Brooks E.D.
        • Chang J.Y.
        Time to abandon single-site irradiation for inducing abscopal effects.
        Nat Rev Clin Oncol. 2019; 16: 123-135https://doi.org/10.1038/s41571-018-0119-7
        • Bos P.D.
        • Plitas G.
        • Rudra D.
        • Lee S.Y.
        • Rudensky A.Y.
        Transient regulatory T cell ablation deters oncogene-driven breast cancer and enhances radiotherapy.
        J Exp Med. 2013; 210: 2435-2466https://doi.org/10.1084/jem.20130762
        • Cassetta L.
        • Kitamura T.
        Macrophage targeting: opening new possibilities for cancer immunotherapy.
        Immunology. 2018; 155: 285-293https://doi.org/10.1111/imm.12976
        • Fleming V.
        • Groth C.
        • Altevogt P.
        • Umansky V.
        • Nagibin V.
        • Weber R.
        • et al.
        Targeting myeloid-derived suppressor cells to bypass tumour-induced immunosuppression.
        Front Immunol. 2018; 9https://doi.org/10.3389/fimmu.2018.00398
        • Tumeh P.C.
        • Harview C.L.
        • Yearley J.H.
        • Shintaku I.P.
        • Taylor E.J.M.
        • Robert L.
        • et al.
        PD-1 blockade induces responses by inhibiting adaptive immune resistance.
        Nature. 2014; 515: 568-571https://doi.org/10.1038/nature13954
        • Andrews M.C.
        • Wargo J.A.
        Immunotherapy resistance: the answers lie ahead - not in front - of us.
        J Immunother Cancer. 2017; 5https://doi.org/10.1186/s40425-017-0212-y
        • Sharma P.
        • Allison J.P.
        The future of immune checkpoint therapy.
        Science. 2015; 348 (80-): 56-61https://doi.org/10.1126/science.aaa8172
        • Vonderheide R.H.
        • Domchek S.M.
        • Clark A.S.
        Immunotherapy for breast cancer: what are we missing?.
        Clin Canc Res. 2017; 23: 2640-2646https://doi.org/10.1158/1078-0432.CCR-16-2569
        • Dewan M.Z.
        • Galloway A.E.
        • Kawashima N.
        • Dewyngaert J.K.
        • Babb J.S.
        • Formenti S.C.
        • et al.
        Fractionated but not single-dose radiotherapy induces an immune-mediated abscopal effect when combined with anti-CTLA-4 antibody.
        Clin Canc Res. 2009; 15: 5379-5388https://doi.org/10.1158/1078-0432.CCR-09-0265
        • Demaria S.
        • Kawashima N.
        • Yang A.M.
        • Devitt M.L.
        • Babb J.S.
        • Allison J.P.
        • et al.
        Immune-mediated inhibition of metastases after treatment with local radiation and CTLA-4 blockade in a mouse model of breast cancer.
        Clin Canc Res. 2005; 11: 728-734
        • Herrera F.G.
        • Bourhis J.
        • Coukos G.
        Radiotherapy combination opportunities leveraging immunity for the next oncology practice.
        CA Cancer J Clin. 2017; 67: 65-85https://doi.org/10.3322/caac.21358
        • Weichselbaum R.R.
        • Liang H.
        • Deng L.
        • Fu Y.-X.
        Radiotherapy and immunotherapy: a beneficial liaison?.
        Nat Rev Clin Oncol. 2017; 14: 365-379https://doi.org/10.1038/nrclinonc.2016.211
        • Wang X.
        • Ding J.
        • Meng L.H.
        PI3K isoform-selective inhibitors: next-generation targeted cancer therapies.
        Acta Pharmacol Sin. 2015; 36: 1170-1176https://doi.org/10.1038/aps.2015.71
        • Ali K.
        • Soond D.R.
        • Piñeiro R.
        • Hagemann T.
        • Pearce W.
        • Lim E.L.
        • et al.
        Inactivation of PI(3)K p110δ breaks regulatory T-cell-mediated immune tolerance to cancer.
        Nature. 2014; 510: 407-411https://doi.org/10.1038/nature13444
        • De Henau O.
        • Rausch M.
        • Winkler D.
        • Campesato L.F.
        • Liu C.
        • Cymerman D.H.
        • et al.
        Overcoming resistance to checkpoint blockade therapy by targeting PI3Kγ in myeloid cells.
        Nature. 2016; 539: 443-447https://doi.org/10.1038/nature20554
        • Tanaskovic O.
        • Verga Falzacappa M.V.
        • Pelicci P.G.
        Human cord blood (hCB)-CD34+ humanised mice fail to reject human acute myeloid leukemia cells.
        PLoS One. 2019; 14e0217345https://doi.org/10.1371/journal.pone.0217345
        • Jassal B.
        • Matthews L.
        • Viteri G.
        • Gong C.
        • Lorente P.
        • Fabregat A.
        • et al.
        The reactome pathway knowledgebase.
        Nucleic Acids Res. 2020; https://doi.org/10.1093/nar/gkz1031
        • Nishimura D.
        BioCarta.
        Biotech Softw Internet Rep. 2001; https://doi.org/10.1089/152791601750294344
        • Newman A.M.
        • Steen C.B.
        • Liu C.L.
        • Gentles A.J.
        • Chaudhuri A.A.
        • Scherer F.
        • et al.
        Determining cell type abundance and expression from bulk tissues with digital cytometry.
        Nat Biotechnol. 2019; https://doi.org/10.1038/s41587-019-0114-2
        • Yong H.J.
        • Choi Y.
        • Joo H.K.
        • Chul W.K.
        • Jae M.J.
        • Dong S.L.
        • et al.
        Immune response to firefly luciferase as a naked DNA.
        Cancer Biol Ther. 2007; https://doi.org/10.4161/cbt.6.5.4005
        • Becht E.
        • Giraldo N.A.
        • Lacroix L.
        • Buttard B.
        • Elarouci N.
        • Petitprez F.
        • et al.
        Estimating the population abundance of tissue-infiltrating immune and stromal cell populations using gene expression.
        Genome Biol. 2016; https://doi.org/10.1186/s13059-016-1070-5
        • Bilanges B.
        • Posor Y.
        • Vanhaesebroeck B.
        PI3K isoforms in cell signalling and vesicle trafficking.
        Nat Rev Mol Cell Biol. 2019; https://doi.org/10.1038/s41580-019-0129-z
        • Motz G.T.
        • Santoro S.P.
        • Wang L.P.
        • Garrabrant T.
        • Lastra R.R.
        • Hagemann I.S.
        • et al.
        Tumour endothelium FasL establishes a selective immune barrier promoting tolerance in tumours.
        Nat Med. 2014; 20: 607-615https://doi.org/10.1038/nm.3541
        • Zhu J.
        • Powis De Tenbossche C.G.
        • Cané S.
        • Colau D.
        • Van Baren N.
        • Lurquin C.
        • et al.
        Resistance to cancer immunotherapy mediated by apoptosis of tumour-infiltrating lymphocytes.
        Nat Commun. 2017; 8https://doi.org/10.1038/s41467-017-00784-1
        • Champiat S.
        • Dercle L.
        • Ammari S.
        • Massard C.
        • Hollebecque A.
        • Postel-Vinay S.
        • et al.
        Hyperprogressive disease is a new pattern of progression in cancer patients treated by anti-PD-1/PD-L1.
        Clin Canc Res. 2017; 23: 1920-1928https://doi.org/10.1158/1078-0432.CCR-16-1741
        • Champiat S.
        • Ferrara R.
        • Massard C.
        • Besse B.
        • Marabelle A.
        • Soria J.C.
        • et al.
        Hyperprogressive disease: recognizing a novel pattern to improve patient management.
        Nat Rev Clin Oncol. 2018; 15: 748-762https://doi.org/10.1038/s41571-018-0111-2
        • Kaneda M.M.
        • Messer K.S.
        • Ralainirina N.
        • Li H.
        • Leem C.J.
        • Gorjestani S.
        • et al.
        PI3Kγ is a molecular switch that controls immune suppression.
        Nature. 2016; 539: 437-442https://doi.org/10.1038/nature19834
        • Kalbasi A.
        • Ribas A.
        Tumour-intrinsic resistance to immune checkpoint blockade.
        Nat Rev Immunol. 2019; https://doi.org/10.1038/s41577-019-0218-4
        • Karam S.D.
        • Raben D.
        Radioimmunotherapy for the treatment of head and neck cancer.
        Lancet Oncol. 2019; 20: e404-e416https://doi.org/10.1016/s1470-2045(19)30306-7
        • Lippitz B.E.
        • Harris R.A.
        A translational concept of immuno-radiobiology.
        Radiother Oncol. 2019; 140: 116-124https://doi.org/10.1016/j.radonc.2019.06.001
        • Demaria S.
        • Coleman C.N.
        • Formenti S.C.
        Radiotherapy: changing the Game in Immunotherapy the promise of radiotherapy as a partner for immunotherapy.
        Trends Cancer. 2017; 2: 286-294https://doi.org/10.1016/j.trecan.2016.05.002
        • Ngwa W.
        • Irabor O.C.
        • Schoenfeld J.D.
        • Hesser J.
        • Demaria S.
        • Formenti S.C.
        Using immunotherapy to boost the abscopal effect.
        Nat Rev Cancer. 2018; 18: 313-322https://doi.org/10.1038/nrc.2018.6
        • Vanpouille-Box C.
        • Alard A.
        • Aryankalayil M.J.
        • Sarfraz Y.
        • Diamond J.M.
        • Schneider R.J.
        • et al.
        DNA exonuclease Trex1 regulates radiotherapy-induced tumour immunogenicity.
        Nat Commun. 2017; 8: 15618https://doi.org/10.1038/ncomms15618
        • Zhang X.
        • Niedermann G.
        Abscopal effects with hypofractionated schedules extending into the effector phase of the tumour-specific T-cell response.
        Int J Radiat Oncol Biol Phys. 2018; https://doi.org/10.1016/j.ijrobp.2018.01.094
        • Barth R.J.
        • Mule J.J.
        • Spiess P.J.
        • Rosenberg S.A.
        Interferon γ and tumour necrosis factor have a role in tumour regressions mediated by murine CD8+ tumour-infiltrating lymphocytes.
        J Exp Med. 1991; 173: 647-658https://doi.org/10.1084/jem.173.3.647
        • Seifert L.
        • Werba G.
        • Tiwari S.
        • Giao Ly N.N.
        • Nguy S.
        • Alothman S.
        • et al.
        Radiation therapy induces macrophages to suppress T-cell responses against pancreatic tumours in mice.
        Gastroenterology. 2016; 150: 1659-1672.e5https://doi.org/10.1053/j.gastro.2016.02.070
        • Thorpe L.M.
        • Yuzugullu H.
        • Zhao J.J.
        PI3K in cancer: divergent roles of isoforms, modes of activation and therapeutic targeting.
        Nat Rev Cancer. 2015; 15: 7-24https://doi.org/10.1038/nrc3860
        • Aran D.
        • Sirota M.
        • Butte A.J.
        Systematic pan-cancer analysis of tumour purity.
        Nat Commun. 2015; 6: 1-12https://doi.org/10.1038/ncomms9971
        • Aran D.
        • Hu Z.
        • Butte A.J.
        xCell: digitally portraying the tissue cellular heterogeneity landscape.
        Genome Biol. 2017; 18: 1-14https://doi.org/10.1186/s13059-017-1349-1