Progesterone promotes immunomodulation and tumor development in the murine mammary gland

J Immunother Cancer. 2021 May;9(5):e001710. doi: 10.1136/jitc-2020-001710.

Abstract

Background: Clinical studies have linked usage of progestins (synthetic progesterone [P4]) to breast cancer risk. However, little is understood regarding the role of native P4, signaling through the progesterone receptor (PR), in breast tumor formation. Recently, we reported a link between PR and immune signaling pathways, showing that P4/PR can repress type I interferon signaling pathways. Given these findings, we sought to investigate whether P4/PR drive immunomodulation in the mammary gland and promote tumor formation.

Methods: To determine the effect of P4 on immune cell populations in the murine mammary gland, mice were treated with P4 or placebo pellets for 21 days. Immune cell populations in the mammary gland, spleen, and inguinal lymph nodes were subsequently analyzed by flow cytometry. To assess the effect of PR overexpression on mammary gland tumor development as well as immune cell populations in the mammary gland, a transgenic mouse model was used in which PR was overexpressed throughout the entire mouse. Immune cell populations were assessed in the mammary glands, spleens, and inguinal lymph nodes of 6-month-old transgenic and control mice by flow cytometry. Transgenic mice were also monitored for mammary gland tumor development over a 2-year time span. Following development of mammary gland tumors, immune cell populations in the tumors and spleens of transgenic and control mice were analyzed by flow cytometry.

Results: We found that mice treated with P4 exhibited changes in the mammary gland indicative of an inhibited immune response compared with placebo-treated mice. Furthermore, transgenic mice with PR overexpression demonstrated decreased numbers of immune cell populations in their mammary glands, lymph nodes, and spleens. On long-term monitoring, we determined that multiparous PR-overexpressing mice developed significantly more mammary gland tumors than control mice. Additionally, tumors from PR-overexpressing mice contained fewer infiltrating immune cells. Finally, RNA sequencing analysis of tumor samples revealed that immune-related gene signatures were lower in tumors from PR-overexpressing mice as compared with control mice.

Conclusion: Together, these findings offer a novel mechanism of P4-driven mammary gland tumor development and provide rationale in investigating the usage of antiprogestin therapies to promote immune-mediated elimination of mammary gland tumors.

Keywords: adaptive immunity; breast neoplasms; immunomodulation; lymphocytes; tumor microenvironment; tumor-infiltrating.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptive Immunity / drug effects
  • Animals
  • Breast Neoplasms / chemically induced*
  • Breast Neoplasms / immunology
  • Breast Neoplasms / metabolism
  • Breast Neoplasms / pathology
  • Cell Line, Tumor
  • Cell Transformation, Neoplastic / chemically induced*
  • Cell Transformation, Neoplastic / immunology
  • Cell Transformation, Neoplastic / metabolism
  • Cell Transformation, Neoplastic / pathology
  • Drug Implants
  • Female
  • Galectin 4 / genetics
  • Galectin 4 / metabolism
  • Immunity, Innate / drug effects
  • Lymphocytes, Tumor-Infiltrating / drug effects
  • Lymphocytes, Tumor-Infiltrating / immunology
  • Lymphocytes, Tumor-Infiltrating / metabolism
  • Mammary Glands, Animal / drug effects*
  • Mammary Glands, Animal / immunology
  • Mammary Glands, Animal / metabolism
  • Mammary Glands, Animal / pathology
  • Mice, Transgenic
  • Ovariectomy
  • Progesterone / administration & dosage*
  • Receptors, Progesterone / agonists*
  • Receptors, Progesterone / genetics
  • Receptors, Progesterone / metabolism
  • Signal Transduction
  • Time Factors
  • Tumor Burden / drug effects
  • Tumor Escape / drug effects*
  • Tumor Microenvironment / immunology*

Substances

  • Drug Implants
  • Galectin 4
  • Receptors, Progesterone
  • Progesterone