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Histone deacetylase inhibition promotes intratumoral CD8+ T-cell responses, sensitizing murine breast tumors to anti-PD1

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Abstract

Histone deacetylase (HDAC) inhibitors impair tumor cell proliferation and alter gene expression. However, the impact of these changes on anti-tumor immunity is poorly understood. Here, we showed that the class I HDAC inhibitor, entinostat (ENT), promoted the expression of immune-modulatory molecules, including MHCII, costimulatory ligands, and chemokines on murine breast tumor cells in vitro and in vivo. ENT also impaired tumor growth in vivo—an effect that was dependent on both CD8+ T cells and IFNγ. Moreover, ENT promoted intratumoral T-cell clonal expansion and enhanced their functional activity. Importantly, ENT sensitized normally unresponsive tumors to the effects of PD1 blockade, predominantly through increases in T-cell proliferation. Our findings suggest that class I HDAC inhibitors impair tumor growth by enhancing the proliferative and functional capacity of CD8+ T cells and by sensitizing tumor cells to T-cell recognition.

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Abbreviations

ATCC:

American-Type Culture Collection

CIITA:

Class II transcriptional activator

ENT:

Entinostat

GZB:

Granzyme B

HAT:

Histone acetyltransferase

HDAC:

Histone deacetylase

MHCII:

MHC class II

PAN:

Panobinostat

TIL:

Tumor-infiltrating lymphocyte

TNBC:

Triple negative breast cancer

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Acknowledgements

The authors would like to thank Uma Mudunuru and Scott Simpler for animal husbandry, and Eddy Yang and Debbie Della Manna of the NanoString Laboratory and Sagar Hanamanthu of the Comprehensive Flow Cytometry Core for lending respective expertise.

Funding

This work was supported by the University of Alabama at Birmingham Comprehensive Cancer Center (P30 CA013148), National Institutes of Health grant CA216234, and by the Breast Cancer Research Foundation of Alabama.

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Authors and Affiliations

  1. Department of Medicine, Division of Clinical Immunology and Rheumatology, University of Alabama at Birmingham, 1720 2nd AVE S, Birmingham, AL, 35294, USA

    Tyler R. McCaw, Mingyong Liu & Troy D. Randall

  2. Department of Radiation Oncology, University of Alabama at Birmingham, Birmingham, AL, USA

    Mei Li & Donald J. Buchsbaum

  3. HudsonAlpha Institute for Biotechnology, Huntsville, AL, USA

    Dmytro Starenki & Sara J. Cooper

  4. Department of Obstetrics and Gynecology, Division of Gynecology Oncology, University of Alabama at Birmingham, Birmingham, AL, USA

    Rebecca C. Arend

  5. Department of Medicine, Division of Hematology and Oncology, University of Alabama at Birmingham, Birmingham, AL, USA

    Andres Forero

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Contributions

TRM designed, performed, and interpreted experiments and wrote the manuscript. ML designed, performed, and interpreted experiments. DS performed TCR repertoire sequencing and analysis. SJC performed TCR repertoire sequencing and analysis. ML designed, performed, and interpreted experiments. RCA designed and interpreted experiments and edited the manuscript. AF designed and interpreted experiments and edited the manuscript. DJB designed and interpreted experiments and edited the manuscript. TDR designed and interpreted experiments and edited the manuscript.

Corresponding author

Correspondence to Troy D. Randall.

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Conflict of interest

The authors declare that they have no conflict of interest.

Research involving human participants and/or animals

All procedures involving animals were performed in accordance with the guidelines of the National Research Council (United States) Committee for the Update of the Guide for the Care and Use of Laboratory Animals and were approved by the University of Alabama at Birmingham Institutional Animal Care and Use Committee (IACUC) in protocol 09854.

Animal source

BALB/c mice were purchased from Charles River Laboratories International, Inc. BALB/c.scid mice (CBySmn.CB17-PrkdcscidIJ) were purchased from The Jackson Laboratory.

Cell line authentication

TS/A murine mammary adenocarcinoma cells were provided by Dr. Roberto S. Accolla, Department of Clinical and Biological Sciences, University of Insubria, Italy. TS/A cells were obtained at passage 22 and passaged two times prior to freezing archival samples. 4T1 cells were purchased from the American Type Culture Collection (ATCC). Samples of 4T1 and TS/A cells were authenticated by IDEXX BioAnalytics: case #23272-2019. All cell lines tested negative for mycoplasma (and 13 other mouse pathogens) via PCR performed by Charles River Research Animal Diagnostic Services.

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McCaw, T.R., Li, M., Starenki, D. et al. Histone deacetylase inhibition promotes intratumoral CD8+ T-cell responses, sensitizing murine breast tumors to anti-PD1. Cancer Immunol Immunother 68, 2081–2094 (2019). https://doi.org/10.1007/s00262-019-02430-9

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