Abstract P2-03-09: Autocrine motility factor signaling pathway promotes aggressive behavior and migration in breast cancer

Abstract

Abstract Background: Autocrine motility factor (AMF) is secreted by cancer cells and acts in an autocrine or paracrine fashion to bind to its receptor, AMFR, at the cell surface of tumor cells. The AMF-AMFR pathway has been shown to promote proliferation, anti-apoptosis, motility and migration, invasion, and metastasis pathways in various cancers. However, our understanding of the AMF-AMFR pathway in breast cancer is limited. We propose that the AMF signaling pathway is an unexplored mechanism for breast cancer tumor aggression and its receptor, AMFR, may be a potential therapeutic target. Methods: Tumor tissue obtained from consented female (n = 31) and male (n = 1) patients were analyzed by the Affymetrix OncoscanTM genome-wide microarray platform and examined for somatic copy-number alterations (SCNAs) of AMFR. cBioPortal was used to investigate SCNA of AMFR and gene expression of AMF on primary breast cancer tumors from METABRIC (n = 1,784) and TCGA Pan-Cancer Atlas (n = 981) datasets. In vitro, AMF and AMFR gene expression in luminal A (MCF-7) and triple-negative breast cancer (MDA-MB-231) cell lines were assessed by qPCR. Cell migration assays were performed on MDA-MB-231 cells to investigate their migration towards AMF with AMFR present or knocked down by siRNA. Results: Microarray analysis of 32 tumors revealed that a single-copy loss of AMFR occurred 79% of the time in luminal A tumors (n = 19); 67% of the time in luminal B tumors (n = 6); 33% of the time in ER+, PR+, HER2+ tumors (n = 3); and 0% of the time in triple-negative breast cancer (TNBC) tumors (n = 4), suggesting that the loss of AMFR results in less aggressive tumors that have good overall prognosis. To extend our findings to a larger patient cohort, SCNA analysis of the METABRIC and TCGA Pan-Cancer Atlas datasets revealed that single-copy loss of AMFR occurred in 64.53% and 68.14% of luminal A tumors, 58.51% and 59.90% of luminal B tumors, 23.62% and 48.72% of HER2 overexpression tumors, and 29.80% and 39.77% of TNBC tumors, respectively. Therefore, AMFR appears most frequently deleted in the tumor genomes of good prognosis breast cancer molecular subtypes (luminal tumors). Gene expression analysis of AMF in the METABRIC (using z-scores) and TCGA Pan-Cancer Atlas (using batch-normalized numbers) datasets revealed median mRNA expressions of -0.3351 and 4862 in luminal A tumors, -0.05415 and 5841 in luminal B tumors, 0.5758 and 9390 in HER2 overexpression tumors, and 0.754 and 8798 in TNBC tumors, respectively, suggesting that the AMF-AMFR pathway is more active in aggressive breast cancers. Similarly, we observed that AMF and AMFR are transcriptionally overexpressed by 7-fold and 16-fold, respectively, in the TNBC cell line MDA-MB-231 compared to the luminal A breast cancer cell line MCF-7. When AMFR is knocked down in MDA-MB-231 cells, focused migration towards AMF is abolished. Conclusion: AMF-AMFR pathway activity correlates with aggressive cancer cell behavior and enhanced migration in breast cancer. Single-copy loss of AMFR in tumor genomes is associated with less aggressive tumors with better overall prognosis. AMFR may be an attractive therapeutic target. Citation Format: Liu C, Price JD, Maroni D, Stevens JM, VanDyke AZ, Althof PA, Mondal P, Sanmann JN, Thayer SP. Autocrine motility factor signaling pathway promotes aggressive behavior and migration in breast cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P2-03-09.