Mammary Carcinoma Cell Derived Cyclooxygenase 2 Suppresses Tumor Immune Surveillance by Enhancing Intratumoral Immune Checkpoint Activity

Breast Cancer Research

Emer M Smyth¹  Robert H Vonderheide³  Victoire Ndong¹  Rebecca A Evans²  Edward P Chen¹  Nune Markosyan¹ 

[1] Institute for Translational Medicine and Therapeutics, Smilow Center forTranslational Research, 3400 Civic Center Blvd, Philadelphia, PA 19104, USA;Abramson Family Cancer Research Institute, 421 Curie Blvd, Philadelphia, PA 19104,USA;Department of Medicine, Perelman School of Medicine at the University ofPennsylvania, 415 Curie Blvd, Philadelphia, PA 19104, USA

关键词: PD-L1;    cytotoxic immune cells;    tumor microenvironment;    immune modulation;    breast cancer;   

Others  :  793692
DOI  :  10.1186/bcr3469

 received in 2012-12-10, accepted in 2013-07-08,  发布年份 2013

【 摘 要 】

Introduction

Systemic inhibition of the inflammatory enzyme cyclooxygenase (COX) 2 decreases the risk of breast cancer and its recurrence. However, the biology of COX-2 in the multicellular tumor microenvironment is poorly defined.

Methods

Mammary tumor onset and multiplicity were examined in ErbB2 transgenic mice that were deficient in mammary epithelial cell COX-2 (COX-2^MECKO) compared to wild type (WT) mice.

Tumors were analyzed, by real time PCR, immune-staining and flow cytometry, for proliferation, apoptosis, angiogenesis and immune microenvironment. Lentiviral shRNA delivery was used to knock down (KD) COX-2 in ErbB2-transformed mouse breast cancer cells (COX-2KD), and growth as orthotopic tumors was examined in syngenic recipient mice, with or without depletion of CD8⁺ immune cells.

Results

Mammary tumor onset was delayed, and multiplicity halved, in COX-2^MECKO mice compared to WT. COX-2^MECKO tumors showed decreased expression of Ki67, a proliferation marker, as well as reduced VEGFA, its receptor VEGFR2, endothelial NOS and the vascular endothelial marker CD31, indicating reduced tumor vascularization. COX-2^MECKO tumors contained more CD4⁺ T helper (T_h) cells and CD8⁺ cytotoxic immune cells (CTL) consistent with increased immune surveillance. The ratio of T_h markers Tbet (T_h1) to GATA3 (T_h2) was higher, and levels of Retnla, a M2 macrophage marker, lower, in COX-2^MECKO tumor infiltrating leukocytes compared to WT, suggesting a prevalence of pro-immune T_h1 over immune suppressive T_h2 lymphocytes, and reduced macrophage polarization to the immune suppressive M2 phenotype. Enhanced immune surveillance in COX-2^MECKO tumors was coincident with increased intratumoral CXCL9, a T cell chemoattractant, and decreased expression of T lymphocyte co-inhibitory receptors CTLA4 and PD-1, as well as PD-L1, the ligand for PD-1. PD-L1 was also decreased in IFNγ-treated COX-2KD mouse mammary cancer cells in vitroand, compared to control cells, growth of COX-2KD cells as orthotopic tumors in immune competent mice was markedly suppressed. However, robust growth of COX-2KD tumor cells was evident when recipients were depleted of CD8⁺cells.

Conclusions

The data strongly support that, in addition to its angiogenic function, tumor cell COX-2 suppresses intratumoral cytotoxic CD8⁺ immune cell function, possibly through upregulation of immune checkpoints, thereby contributing to tumor immune escape. COX-2 inhibition may be clinically useful to augment breast cancer immunotherapy.

【 授权许可】

   
2013 Markosyan et al.; licensee BioMed Central Ltd.

【 图 表 】

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