Resistance to radiotherapy and PD-L1 blockade is mediated by TIM-3 upregulation and regulatory T-cell infiltration

Ayman Oweida, Mohammad K. Hararah, Andy Phan, David Binder, Shilpa Bhatia, Shelby Lennon, Sanjana Bukkapatnam, Benjamin Van Court, Nomin Uyanga, Laurel Darragh, Hyun Min Kim, David Raben, Aik-Choon Tan, Lynn Heasley, Eric Clambey, Raphael Nemenoff, Sana D. Karam

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Purpose: Radiotherapy (RT) can transform the immune landscape and render poorly immunogenic tumors sensitive to PD-L1 inhibition. Here, we established that the response to combined RT and PD-L1 inhibition is transient and investigated mechanisms of resistance. Experimental Design: Mechanisms of resistance to RT and PD-L1 blockade were investigated in orthotopic murine head and neck squamous cell carcinoma (HNSCC) tumors using mass cytometry and whole-genome sequencing. Mice were treated with anti-PD-L1 or anti-TIM-3 alone and in combination with and without RT. Tumor growth and survival were assessed. Flow cytometry was used to assess phenotypic and functional changes in intratumoral T-cell populations. Depletion of regulatory T cells (Treg) was performed using anti- CD25 antibody. Results: We show that the immune checkpoint receptor, TIM-3, is upregulated on CD8 T cells and Tregs in tumors treated with RT and PD-L1 blockade. Treatment with anti- TIM-3 concurrently with anti-PD-L1 and RT led to significant tumor growth delay, enhanced T-cell cytotoxicity, decreased Tregs, and improved survival in orthotopic models of HNSCC. Despite this treatment combination, the response was not durable, and analysis of relapsed tumors revealed resurgence of Tregs. Targeted Treg depletion, however, restored antitumor immunity in mice treated with RT and dual immune checkpoint blockade and resulted in tumor rejection and induction of immunologic memory. Conclusions: These data reveal multiple layers of immune regulation that can promote tumorigenesis and the therapeutic potential of sequential targeting to overcome tumor resistance mechanisms. We propose that targeted Treg inhibitors may be critical for achieving durable tumor response with combined radiotherapy and immunotherapy.

Original languageEnglish
Pages (from-to)5368-5380
Number of pages13
JournalClinical Cancer Research
Volume24
Issue number21
DOIs
Publication statusPublished - 2018 Nov 1
Externally publishedYes

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Regulatory T-Lymphocytes
Up-Regulation
Radiotherapy
Neoplasms
T-Lymphocytes
Immunologic Memory
Growth
Immunotherapy
Anti-Idiotypic Antibodies
Immunity
Flow Cytometry
Carcinogenesis
Research Design
Therapeutics
Genome

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Oweida, A., Hararah, M. K., Phan, A., Binder, D., Bhatia, S., Lennon, S., ... Karam, S. D. (2018). Resistance to radiotherapy and PD-L1 blockade is mediated by TIM-3 upregulation and regulatory T-cell infiltration. Clinical Cancer Research, 24(21), 5368-5380. https://doi.org/10.1158/1078-0432.CCR-18-1038

Resistance to radiotherapy and PD-L1 blockade is mediated by TIM-3 upregulation and regulatory T-cell infiltration. / Oweida, Ayman; Hararah, Mohammad K.; Phan, Andy; Binder, David; Bhatia, Shilpa; Lennon, Shelby; Bukkapatnam, Sanjana; Van Court, Benjamin; Uyanga, Nomin; Darragh, Laurel; Kim, Hyun Min; Raben, David; Tan, Aik-Choon; Heasley, Lynn; Clambey, Eric; Nemenoff, Raphael; Karam, Sana D.

In: Clinical Cancer Research, Vol. 24, No. 21, 01.11.2018, p. 5368-5380.

Research output: Contribution to journalArticle

Oweida, A, Hararah, MK, Phan, A, Binder, D, Bhatia, S, Lennon, S, Bukkapatnam, S, Van Court, B, Uyanga, N, Darragh, L, Kim, HM, Raben, D, Tan, A-C, Heasley, L, Clambey, E, Nemenoff, R & Karam, SD 2018, 'Resistance to radiotherapy and PD-L1 blockade is mediated by TIM-3 upregulation and regulatory T-cell infiltration', Clinical Cancer Research, vol. 24, no. 21, pp. 5368-5380. https://doi.org/10.1158/1078-0432.CCR-18-1038
Oweida, Ayman ; Hararah, Mohammad K. ; Phan, Andy ; Binder, David ; Bhatia, Shilpa ; Lennon, Shelby ; Bukkapatnam, Sanjana ; Van Court, Benjamin ; Uyanga, Nomin ; Darragh, Laurel ; Kim, Hyun Min ; Raben, David ; Tan, Aik-Choon ; Heasley, Lynn ; Clambey, Eric ; Nemenoff, Raphael ; Karam, Sana D. / Resistance to radiotherapy and PD-L1 blockade is mediated by TIM-3 upregulation and regulatory T-cell infiltration. In: Clinical Cancer Research. 2018 ; Vol. 24, No. 21. pp. 5368-5380.
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T1 - Resistance to radiotherapy and PD-L1 blockade is mediated by TIM-3 upregulation and regulatory T-cell infiltration

AU - Oweida, Ayman

AU - Hararah, Mohammad K.

AU - Phan, Andy

AU - Binder, David

AU - Bhatia, Shilpa

AU - Lennon, Shelby

AU - Bukkapatnam, Sanjana

AU - Van Court, Benjamin

AU - Uyanga, Nomin

AU - Darragh, Laurel

AU - Kim, Hyun Min

AU - Raben, David

AU - Tan, Aik-Choon

AU - Heasley, Lynn

AU - Clambey, Eric

AU - Nemenoff, Raphael

AU - Karam, Sana D.

PY - 2018/11/1

Y1 - 2018/11/1

N2 - Purpose: Radiotherapy (RT) can transform the immune landscape and render poorly immunogenic tumors sensitive to PD-L1 inhibition. Here, we established that the response to combined RT and PD-L1 inhibition is transient and investigated mechanisms of resistance. Experimental Design: Mechanisms of resistance to RT and PD-L1 blockade were investigated in orthotopic murine head and neck squamous cell carcinoma (HNSCC) tumors using mass cytometry and whole-genome sequencing. Mice were treated with anti-PD-L1 or anti-TIM-3 alone and in combination with and without RT. Tumor growth and survival were assessed. Flow cytometry was used to assess phenotypic and functional changes in intratumoral T-cell populations. Depletion of regulatory T cells (Treg) was performed using anti- CD25 antibody. Results: We show that the immune checkpoint receptor, TIM-3, is upregulated on CD8 T cells and Tregs in tumors treated with RT and PD-L1 blockade. Treatment with anti- TIM-3 concurrently with anti-PD-L1 and RT led to significant tumor growth delay, enhanced T-cell cytotoxicity, decreased Tregs, and improved survival in orthotopic models of HNSCC. Despite this treatment combination, the response was not durable, and analysis of relapsed tumors revealed resurgence of Tregs. Targeted Treg depletion, however, restored antitumor immunity in mice treated with RT and dual immune checkpoint blockade and resulted in tumor rejection and induction of immunologic memory. Conclusions: These data reveal multiple layers of immune regulation that can promote tumorigenesis and the therapeutic potential of sequential targeting to overcome tumor resistance mechanisms. We propose that targeted Treg inhibitors may be critical for achieving durable tumor response with combined radiotherapy and immunotherapy.

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