Combination of anthracyclines and anti-CD47 therapy inhibit invasive breast cancer growth while preventing cardiac toxicity by regulation of autophagy
- 679 Downloads
A perennial challenge in systemic cytotoxic cancer therapy is to eradicate primary tumors and metastatic disease while sparing normal tissue from off-target effects of chemotherapy. Anthracyclines such as doxorubicin are effective chemotherapeutic agents for which dosing is limited by development of cardiotoxicity. Our published evidence shows that targeting CD47 enhances radiation-induced growth delay of tumors while remarkably protecting soft tissues. The protection of cell viability observed with CD47 is mediated autonomously by activation of protective autophagy. However, whether CD47 protects cancer cells from cytotoxic chemotherapy is unknown.
We tested the effect of CD47 blockade on cancer cell survival using a 2-dimensional high-throughput cell proliferation assay in 4T1 breast cancer cell lines. To evaluate blockade of CD47 in combination with chemotherapy in vivo, we employed the 4T1 breast cancer model and examined tumor and cardiac tissue viability as well as autophagic flux.
Our high-throughput screen revealed that blockade of CD47 does not interfere with the cytotoxic activity of anthracyclines against 4T1 breast cancer cells. Targeting CD47 enhanced the effect of doxorubicin chemotherapy in vivo by reducing tumor growth and metastatic spread by activation of an anti-tumor innate immune response. Moreover, systemic suppression of CD47 protected cardiac tissue viability and function in mice treated with doxorubicin.
Our experiments indicate that the protective effects observed with CD47 blockade are mediated through upregulation of autophagic flux. However, the absence of CD47 in did not elicit a protective effect in cancer cells, but it enhanced macrophage-mediated cancer cell cytolysis. Therefore, the differential responses observed with CD47 blockade are due to autonomous activation of protective autophagy in normal tissue and enhancement immune cytotoxicity against cancer cells.
KeywordsCD47 Autophagy Cytoprotection Breast cancer Cardio-oncology
Activity of maximum concentration
Cluster of differentiation
- CD47 (−)
Concentration response curves
Dulbecco’s modified eagle medium
Damage-associated molecular patterns
Hypoxia-inducible factor 1
Signal regulatory protein alpha
Unfolded protein response
We would like to acknowledge the statistical and editorial assistance of the Wake Forest Clinical and Translational Science Institute (WF CTSI), which is supported by NCATS, National Institutes of Health, through Grant Award Number UL1TR001420.
This work was supported by the NCI Career Development Award-K22 1K22CA181274-01A1 (DSP), Wake Forest Baptist Comprehensive Cancer Center’s NCI Cancer Center Support Grant P30CA012197 (DSP, KLC), the Intramural Research Program of the NIH/NCI (DDR) and The National Center for Advancing Translational Sciences (NCATS) (MF, CJT).
Compliance with ethical standards
Conflict of interest
The authors of this manuscript have no conflicts of interest to report.
Experiments in presented in this manuscript comply with the current laws of the United States of America and institutional research integrity policies. Tissue arrays from developed from tissues of human breast cancer patient sections analyzed in the Laboratory of Pathology National Cancer Institute, under approved protocol by the Institutional Review Board of the National Cancer Institute. Animal studies and procedures were approved by the Internal Animal Care and Use Committee (IACUC) of the NIH Intramural Research Program protocol # LP-012 and the Wake Forest IACUC protocol # A16-085. Other datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
- 11.Early Breast Cancer Trialists’ Collaborative G, Peto R, Davies C, Godwin J, Gray R, Pan HC, Clarke M, Cutter D, Darby S, McGale P et al (2012) Comparisons between different polychemotherapy regimens for early breast cancer: meta-analyses of long-term outcome among 100,000 women in 123 randomised trials. Lancet 379(9814):432–444CrossRefGoogle Scholar
- 12.Coombes RC, Bliss JM, Wils J, Morvan F, Espie M, Amadori D, Gambrosier P, Richards M, Aapro M, Villar-Grimalt A et al (1996) Adjuvant cyclophosphamide, methotrexate, and fluorouracil versus fluorouracil, epirubicin, and cyclophosphamide chemotherapy in premenopausal women with axillary node-positive operable breast cancer: results of a randomized trial. The International Collaborative Cancer Group. J Clin Oncol 14(1):35–45CrossRefGoogle Scholar
- 13.Ejlertsen B, Mouridsen HT, Jensen MB, Andersen J, Cold S, Edlund P, Ewertz M, Jensen BB, Kamby C, Nordenskjold B et al (2007) Improved outcome from substituting methotrexate with epirubicin: results from a randomised comparison of CMF versus CEF in patients with primary breast cancer. Eur J Cancer 43(5):877–884CrossRefGoogle Scholar
- 14.Hamilton DH, Matthews Griner L, Keller JM, Hu X, Southall N, Marugan J, David JM, Ferrer M, Palena C (2016) Targeting estrogen receptor signaling with fulvestrant enhances immune and chemotherapy-mediated cytotoxicity of human lung cancer. Clin Cancer ResGoogle Scholar
- 15.Inglese J, Auld DS, Jadhav A, Johnson RL, Simeonov A, Yasgar A, Zheng W, Austin CP (2006) Quantitative high-throughput screening: a titration-based approach that efficiently identifies biological activities in large chemical libraries. Proc Natl Acad Sci USA 103(31):11473–11478CrossRefGoogle Scholar
- 17.Willingham SB, Volkmer JP, Gentles AJ, Sahoo D, Dalerba P, Mitra SS, Wang J, Contreras-Trujillo H, Martin R, Cohen JD et al (2012) The CD47-signal regulatory protein alpha (SIRPa) interaction is a therapeutic target for human solid tumors. Proc Natl Acad Sci USA 109(17):6662–6667CrossRefGoogle Scholar
- 19.Harris LN, Ismaila N, McShane LM, Andre F, Collyar DE, Gonzalez-Angulo AM, Hammond EH, Kuderer NM, Liu MC, Mennel RG et al (2016) Use of biomarkers to guide decisions on adjuvant systemic therapy for women with early-stage invasive breast cancer: american society of clinical oncology clinical practice guideline. J Clin Oncol 34(10):1134–1150CrossRefGoogle Scholar
- 35.Chan KS, Espinosa I, Chao M, Wong D, Ailles L, Diehn M, Gill H, Presti J Jr, Chang HY, van de Rijn M et al (2009) Identification, molecular characterization, clinical prognosis, and therapeutic targeting of human bladder tumor-initiating cells. Proc Natl Acad Sci USA 106(33):14016–14021CrossRefGoogle Scholar
- 36.Soto-Pantoja DR, Isenberg JS, Roberts DD (2011) Therapeutic targeting of CD47 to modulate tissue responses to ischemia and radiation. J Genet Syndr Gene Ther 2(2)Google Scholar
- 38.Rath GM, Schneider C, Dedieu S, Rothhut B, Soula-Rothhut M, Ghoneim C, Sid B, Morjani H, El Btaouri H, Martiny L (2006) The C-terminal CD47/IAP-binding domain of thrombospondin-1 prevents camptothecin- and doxorubicin-induced apoptosis in human thyroid carcinoma cells. Biochimica et biophysica acta 1763(10):1125–1134CrossRefGoogle Scholar
- 42.White E (2007) Role of the metabolic stress responses of apoptosis and autophagy in tumor suppression. In: Ernst Schering Foundation symposium proceedings, pp 23–34Google Scholar