The efficacy of etanercept as anti-breast cancer treatment is attenuated by residing macrophages

Elnaz Shirmohammadi,Seyed-Esmaeil Sadat Ebrahimi,Amir Farshchi,Mona Salimi

doi:10.1186/s12885-020-07228-y

Abstract

BackgroundInteraction between microenvironment and breast cancer cells often is not considered at the early stages of drug development leading to failure of many drugs at later clinical stages. Etanercept is a TNF-alpha inhibitor that has been investigated for potential antitumor effect in breast cancer with conflicting results.MethodsSecretome data on MDA-MB-231 cancer cell-line were from public repositories and subjected to gene enrichment analyses. Since MDA-MB-231 cells secrete high levels of Granulocyte-Monocyte Colony Stimulating Factor, which activates macrophages to promote tumor growth, the effect of macrophage co-culturing on anticancer efficacy of Etanercept in breast cancer was evaluated using the Boolean network modeling and in vitro experiments including invasion, cell cycle, Annexin PI, and tetrazolium based viability assays and NFKB activity.ResultsThe secretome profile of MDA-MB-231 cells was similar to the expression of genes following treatment of breast cancer cells with TNF-α. Accordingly, inhibition of TNF-α by Etanercept decreased MDA-MB-231 cell survival, induced apoptosis and cell cycle arrest in vitro and inhibited NFKB activation. The inhibitory effect of Etanercept on cell viability, cell cycle progression, invasion and induction of apoptosis decreased following co-culturing of the cancer cells with macrophages. The Boolean network modeling of the changes in the dynamics of intracellular signaling pathways revealed NFKB activation by secretome of macrophages, leading to a decreased efficacy of Etanercept, suggesting NFKB inhibition as an alternative approach to inhibit cancer cell growth in the presence of macrophage crosstalk.ConclusionThis study indicates that the effect of Etanercept may be influenced by residing macrophages in tumor microenvironment, and suggests a method to predict the effect of drugs in the presence of stromal cells to guide experimental designs in drug development.

Highlights

Interaction between microenvironment and breast cancer cells often is not considered at the early stages of drug development leading to failure of many drugs at later clinical stages
Using up-regulated signatures induced by ligand perturbations in Library of Integrated Network of Cellular Signaling (LINCS) L1000 module within Enrichr, the list of secreted cytokines from MDA-MB-231 was found to be statistically mimicked when breast cancer cells were treated with IL1 and TNF-α (Table 1)
These results suggested NFKB as being highly active in the invasive breast cancer cells resulting in the secretion of a number of cytokines similar to those secreted by cells treated with TNF-α

Summary

Introduction

Interaction between microenvironment and breast cancer cells often is not considered at the early stages of drug development leading to failure of many drugs at later clinical stages. One of the most influential cytokines in tumor microenvironment is Tumor Necrosis Factor-alpha (TNF-α) [10, 11], long known for its dual effects cancer [12]. Considering the pleiotropic and conflicting effects of TNF-α, various efforts have been made to exploit this cytokine as a therapeutic target in cancer. In this regard, some approaches have used TNF-α administration to tumor tissue to induce cell death due to its necrotizing effects [14]. TNF-α activates NFKB transcription factor along with JNK and p38 in a cellular context-dependent manner [16] This canonical pathway induces the expression of several downstream targets of NFKB leading to the increased invasiveness of cancer cells. A different approach that has attracted a great of attention is the use of TNF-α inhibitors, such as Etanercept, as candidates for cancer biotherapy [17]

Methods

Results

Conclusion