Spontaneous regression of micro-metastases following primary tumor excision: a critical role for primary tumor secretome

Abstract

Numerous case studies have reported spontaneous regression of recognized metastases following primary tumor (PT) excision, but underlying mechanisms are elusive. Here we present a model of metastases regression and latency following PT excision, and identify potential underlying mechanisms. Using MDA-MB-231 HM human breast cancer cells that 5 express highly sensitive luciferase, we were able to monitor early stages of spontaneous metastases development in BALB/c nu/nu mice. Removal of the PT caused marked regression of the smallest micro-metastases, but not of larger metastases, and in vivo supplementation of tumor secretome diminished this regression, suggesting that PTsecreted factors promote early metastatic growth. Correspondingly, cancer cell conditioned 10 medium reduced apoptosis and enhanced MDA-MB-231 HM adhesion in vitro. To identify specific mediating factors, cytokine array and proteomic analysis of MDA-MB-231 HM secretome were conducted. Results identified significant enrichment of angiogenesis, growth factors binding and activity, focal adhesion, metalloprotease regulation, and apoptosis regulation processes. Simultaneous in vivo blockade of four secreted key 15 potential mediators of these processes, IL-8, PDGFaa, Serpin E1 (PAI-1), and MIF, arrested development of micro-metastases in the presence of the PT. Interestingly, using the public TCGA provisional dataset, high protein levels of these four factors were correlated with poor survival in a cohort of lung adenocarcinoma patients. These results demonstrate regression and latency of micro-metastases following PT excision, and a 20 crucial role for PT-secretome in promoting early metastatic stages in MDA-MB-231 HM xenografts. If generalized, such findings can suggest novel approaches to control minimal residual disease during and following PT excision.