Immune microenvironment modulation unmasks therapeutic benefit of radiotherapy and checkpoint inhibition

Jared M. Newton, Aurelie Hanoteau, Hsuan Chen Liu, Angelina Gaspero, Falguni Parikh, Robyn D. Gartrell-Corrado, Thomas D. Hart, Damya Laoui, Jo A. Van Ginderachter, Neeraja Dharmaraj, William C. Spanos, Yvonne Saenger, Simon Young, Andrew G. Sikora

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Background: Immune checkpoint inhibitors (ICIs) for solid tumors, including those targeting programmed cell death 1 (PD-1) and cytotoxic T lymphocyte-Associated antigen 4 (CTLA-4), have shown impressive clinical efficacy, however, most patients do not achieve durable responses. One major therapeutic obstacle is the immunosuppressive tumor immune microenvironment (TIME). Thus, we hypothesized that a strategy combining tumor-directed radiation with TIME immunomodulation could improve ICI response rates in established solid tumors. Methods: Using a syngeneic mouse model of human papillomavirus (HPV)-Associated head and neck cancer, mEER, we developed a maximally effective regimen combining PD-1 and CTLA-4 inhibition, tumor-directed radiation, and two existing immunomodulatory drugs: cyclophosphamide (CTX) and a small-molecule inducible nitric oxide synthase (iNOS) inhibitor, L-n6-(1-iminoethyl)-lysine (L-NIL). We compared the effects of the various combinations of this regimen on tumor growth, overall survival, establishment of immunologic memory, and immunologic changes with flow cytometry and quantitative multiplex immunofluorescence. Results: We found PD-1 and CTLA-4 blockade, and radiotherapy alone or in combination, incapable of clearing established tumors or reversing the unfavorable balance of effector to suppressor cells in the TIME. However, modulation of the TIME with cyclophosphamide (CTX) and L-NIL in combination with dual checkpoint inhibition and radiation led to rejection of over 70% of established mEER tumors and doubled median survival in the B16 melanoma model. Anti-Tumor activity was CD8+ T cell-dependent and led to development of immunologic memory against tumor-Associated HPV antigens. Immune profiling revealed that CTX/L-NIL induced remodeling of myeloid cell populations in the TIME and tumor-draining lymph node and drove subsequent activation and intratumoral infiltration of CD8+ effector T cells. Conclusions: Overall, this study demonstrates that modulation of the immunosuppressive TIME is required to unlock the benefits of ICIs and radiotherapy to induce immunologic rejection of treatment-refractory established solid tumors.

Original languageEnglish (US)
Article number216
JournalJournal for ImmunoTherapy of Cancer
Volume7
Issue number1
DOIs
StatePublished - Aug 13 2019

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Radiotherapy
Tumor Microenvironment
Neoplasms
CTLA-4 Antigen
Therapeutics
Immunologic Memory
Cell Death
Radiation
Immunosuppressive Agents
Cyclophosphamide
T-Lymphocytes
Experimental Melanomas
Survival
Immunomodulation
Nitric Oxide Synthase Type II
Myeloid Cells
Head and Neck Neoplasms
Fluorescent Antibody Technique
Flow Cytometry
Lymph Nodes

Keywords

  • Cyclophosphamide (CTX)
  • Cytotoxic T lymphocyte associated antigen-4 (CTLA-4)
  • Head and neck cancer
  • Human papillomavirus (HPV)
  • Immune checkpoint inhibitors
  • Immunotherapy
  • L-n6-(1-iminoethyl)-lysine (L-NIL)
  • Programmed cell death protein-1 (PD-1)
  • Radiotherapy
  • Tumor immune microenvironment

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Molecular Medicine
  • Oncology
  • Pharmacology
  • Cancer Research

Cite this

Newton, J. M., Hanoteau, A., Liu, H. C., Gaspero, A., Parikh, F., Gartrell-Corrado, R. D., ... Sikora, A. G. (2019). Immune microenvironment modulation unmasks therapeutic benefit of radiotherapy and checkpoint inhibition. Journal for ImmunoTherapy of Cancer, 7(1), [216]. https://doi.org/10.1186/s40425-019-0698-6

Immune microenvironment modulation unmasks therapeutic benefit of radiotherapy and checkpoint inhibition. / Newton, Jared M.; Hanoteau, Aurelie; Liu, Hsuan Chen; Gaspero, Angelina; Parikh, Falguni; Gartrell-Corrado, Robyn D.; Hart, Thomas D.; Laoui, Damya; Van Ginderachter, Jo A.; Dharmaraj, Neeraja; Spanos, William C.; Saenger, Yvonne; Young, Simon; Sikora, Andrew G.

In: Journal for ImmunoTherapy of Cancer, Vol. 7, No. 1, 216, 13.08.2019.

Research output: Contribution to journalArticle

Newton, JM, Hanoteau, A, Liu, HC, Gaspero, A, Parikh, F, Gartrell-Corrado, RD, Hart, TD, Laoui, D, Van Ginderachter, JA, Dharmaraj, N, Spanos, WC, Saenger, Y, Young, S & Sikora, AG 2019, 'Immune microenvironment modulation unmasks therapeutic benefit of radiotherapy and checkpoint inhibition', Journal for ImmunoTherapy of Cancer, vol. 7, no. 1, 216. https://doi.org/10.1186/s40425-019-0698-6
Newton, Jared M. ; Hanoteau, Aurelie ; Liu, Hsuan Chen ; Gaspero, Angelina ; Parikh, Falguni ; Gartrell-Corrado, Robyn D. ; Hart, Thomas D. ; Laoui, Damya ; Van Ginderachter, Jo A. ; Dharmaraj, Neeraja ; Spanos, William C. ; Saenger, Yvonne ; Young, Simon ; Sikora, Andrew G. / Immune microenvironment modulation unmasks therapeutic benefit of radiotherapy and checkpoint inhibition. In: Journal for ImmunoTherapy of Cancer. 2019 ; Vol. 7, No. 1.
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abstract = "Background: Immune checkpoint inhibitors (ICIs) for solid tumors, including those targeting programmed cell death 1 (PD-1) and cytotoxic T lymphocyte-Associated antigen 4 (CTLA-4), have shown impressive clinical efficacy, however, most patients do not achieve durable responses. One major therapeutic obstacle is the immunosuppressive tumor immune microenvironment (TIME). Thus, we hypothesized that a strategy combining tumor-directed radiation with TIME immunomodulation could improve ICI response rates in established solid tumors. Methods: Using a syngeneic mouse model of human papillomavirus (HPV)-Associated head and neck cancer, mEER, we developed a maximally effective regimen combining PD-1 and CTLA-4 inhibition, tumor-directed radiation, and two existing immunomodulatory drugs: cyclophosphamide (CTX) and a small-molecule inducible nitric oxide synthase (iNOS) inhibitor, L-n6-(1-iminoethyl)-lysine (L-NIL). We compared the effects of the various combinations of this regimen on tumor growth, overall survival, establishment of immunologic memory, and immunologic changes with flow cytometry and quantitative multiplex immunofluorescence. Results: We found PD-1 and CTLA-4 blockade, and radiotherapy alone or in combination, incapable of clearing established tumors or reversing the unfavorable balance of effector to suppressor cells in the TIME. However, modulation of the TIME with cyclophosphamide (CTX) and L-NIL in combination with dual checkpoint inhibition and radiation led to rejection of over 70{\%} of established mEER tumors and doubled median survival in the B16 melanoma model. Anti-Tumor activity was CD8+ T cell-dependent and led to development of immunologic memory against tumor-Associated HPV antigens. Immune profiling revealed that CTX/L-NIL induced remodeling of myeloid cell populations in the TIME and tumor-draining lymph node and drove subsequent activation and intratumoral infiltration of CD8+ effector T cells. Conclusions: Overall, this study demonstrates that modulation of the immunosuppressive TIME is required to unlock the benefits of ICIs and radiotherapy to induce immunologic rejection of treatment-refractory established solid tumors.",
keywords = "Cyclophosphamide (CTX), Cytotoxic T lymphocyte associated antigen-4 (CTLA-4), Head and neck cancer, Human papillomavirus (HPV), Immune checkpoint inhibitors, Immunotherapy, L-n6-(1-iminoethyl)-lysine (L-NIL), Programmed cell death protein-1 (PD-1), Radiotherapy, Tumor immune microenvironment",
author = "Newton, {Jared M.} and Aurelie Hanoteau and Liu, {Hsuan Chen} and Angelina Gaspero and Falguni Parikh and Gartrell-Corrado, {Robyn D.} and Hart, {Thomas D.} and Damya Laoui and {Van Ginderachter}, {Jo A.} and Neeraja Dharmaraj and Spanos, {William C.} and Yvonne Saenger and Simon Young and Sikora, {Andrew G.}",
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T1 - Immune microenvironment modulation unmasks therapeutic benefit of radiotherapy and checkpoint inhibition

AU - Newton, Jared M.

AU - Hanoteau, Aurelie

AU - Liu, Hsuan Chen

AU - Gaspero, Angelina

AU - Parikh, Falguni

AU - Gartrell-Corrado, Robyn D.

AU - Hart, Thomas D.

AU - Laoui, Damya

AU - Van Ginderachter, Jo A.

AU - Dharmaraj, Neeraja

AU - Spanos, William C.

AU - Saenger, Yvonne

AU - Young, Simon

AU - Sikora, Andrew G.

PY - 2019/8/13

Y1 - 2019/8/13

N2 - Background: Immune checkpoint inhibitors (ICIs) for solid tumors, including those targeting programmed cell death 1 (PD-1) and cytotoxic T lymphocyte-Associated antigen 4 (CTLA-4), have shown impressive clinical efficacy, however, most patients do not achieve durable responses. One major therapeutic obstacle is the immunosuppressive tumor immune microenvironment (TIME). Thus, we hypothesized that a strategy combining tumor-directed radiation with TIME immunomodulation could improve ICI response rates in established solid tumors. Methods: Using a syngeneic mouse model of human papillomavirus (HPV)-Associated head and neck cancer, mEER, we developed a maximally effective regimen combining PD-1 and CTLA-4 inhibition, tumor-directed radiation, and two existing immunomodulatory drugs: cyclophosphamide (CTX) and a small-molecule inducible nitric oxide synthase (iNOS) inhibitor, L-n6-(1-iminoethyl)-lysine (L-NIL). We compared the effects of the various combinations of this regimen on tumor growth, overall survival, establishment of immunologic memory, and immunologic changes with flow cytometry and quantitative multiplex immunofluorescence. Results: We found PD-1 and CTLA-4 blockade, and radiotherapy alone or in combination, incapable of clearing established tumors or reversing the unfavorable balance of effector to suppressor cells in the TIME. However, modulation of the TIME with cyclophosphamide (CTX) and L-NIL in combination with dual checkpoint inhibition and radiation led to rejection of over 70% of established mEER tumors and doubled median survival in the B16 melanoma model. Anti-Tumor activity was CD8+ T cell-dependent and led to development of immunologic memory against tumor-Associated HPV antigens. Immune profiling revealed that CTX/L-NIL induced remodeling of myeloid cell populations in the TIME and tumor-draining lymph node and drove subsequent activation and intratumoral infiltration of CD8+ effector T cells. Conclusions: Overall, this study demonstrates that modulation of the immunosuppressive TIME is required to unlock the benefits of ICIs and radiotherapy to induce immunologic rejection of treatment-refractory established solid tumors.

AB - Background: Immune checkpoint inhibitors (ICIs) for solid tumors, including those targeting programmed cell death 1 (PD-1) and cytotoxic T lymphocyte-Associated antigen 4 (CTLA-4), have shown impressive clinical efficacy, however, most patients do not achieve durable responses. One major therapeutic obstacle is the immunosuppressive tumor immune microenvironment (TIME). Thus, we hypothesized that a strategy combining tumor-directed radiation with TIME immunomodulation could improve ICI response rates in established solid tumors. Methods: Using a syngeneic mouse model of human papillomavirus (HPV)-Associated head and neck cancer, mEER, we developed a maximally effective regimen combining PD-1 and CTLA-4 inhibition, tumor-directed radiation, and two existing immunomodulatory drugs: cyclophosphamide (CTX) and a small-molecule inducible nitric oxide synthase (iNOS) inhibitor, L-n6-(1-iminoethyl)-lysine (L-NIL). We compared the effects of the various combinations of this regimen on tumor growth, overall survival, establishment of immunologic memory, and immunologic changes with flow cytometry and quantitative multiplex immunofluorescence. Results: We found PD-1 and CTLA-4 blockade, and radiotherapy alone or in combination, incapable of clearing established tumors or reversing the unfavorable balance of effector to suppressor cells in the TIME. However, modulation of the TIME with cyclophosphamide (CTX) and L-NIL in combination with dual checkpoint inhibition and radiation led to rejection of over 70% of established mEER tumors and doubled median survival in the B16 melanoma model. Anti-Tumor activity was CD8+ T cell-dependent and led to development of immunologic memory against tumor-Associated HPV antigens. Immune profiling revealed that CTX/L-NIL induced remodeling of myeloid cell populations in the TIME and tumor-draining lymph node and drove subsequent activation and intratumoral infiltration of CD8+ effector T cells. Conclusions: Overall, this study demonstrates that modulation of the immunosuppressive TIME is required to unlock the benefits of ICIs and radiotherapy to induce immunologic rejection of treatment-refractory established solid tumors.

KW - Cyclophosphamide (CTX)

KW - Cytotoxic T lymphocyte associated antigen-4 (CTLA-4)

KW - Head and neck cancer

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KW - Immune checkpoint inhibitors

KW - Immunotherapy

KW - L-n6-(1-iminoethyl)-lysine (L-NIL)

KW - Programmed cell death protein-1 (PD-1)

KW - Radiotherapy

KW - Tumor immune microenvironment

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