Metastasis Suppressors Regulate the Tumor Microenvironment by Blocking Recruitment of Prometastatic Tumor-Associated Macrophages

Cancer Res. 2015 Oct 1;75(19):4063-73. doi: 10.1158/0008-5472.CAN-14-3394. Epub 2015 Aug 3.


Triple-negative breast cancer (TNBC) patients have the highest risk of recurrence and metastasis. Because they cannot be treated with targeted therapies, and many do not respond to chemotherapy, they represent a clinically underserved group. TNBC is characterized by reduced expression of metastasis suppressors such as Raf kinase inhibitory protein (RKIP), which inhibits tumor invasiveness. Mechanisms by which metastasis suppressors alter tumor cells are well characterized; however, their ability to regulate the tumor microenvironment and the importance of such regulation to metastasis suppression are incompletely understood. Here, we use species-specific RNA sequencing to show that RKIP expression in tumors markedly reduces the number and metastatic potential of infiltrating tumor-associated macrophages (TAM). TAMs isolated from nonmetastatic RKIP(+) tumors, relative to metastatic RKIP(-) tumors, exhibit a reduced ability to drive tumor cell invasion and decreased secretion of prometastatic factors, including PRGN, and shed TNFR2. RKIP regulates TAM recruitment by blocking HMGA2, resulting in reduced expression of numerous macrophage chemotactic factors, including CCL5. CCL5 overexpression in RKIP(+) tumors restores recruitment of prometastatic TAMs and intravasation, whereas treatment with the CCL5 receptor antagonist Maraviroc reduces TAM infiltration. These results highlight the importance of RKIP as a regulator of TAM recruitment through chemokines such as CCL5. The clinical significance of these interactions is underscored by our demonstration that a signature comprised of RKIP signaling and prometastatic TAM factors strikingly separates TNBC patients based on survival outcome. Collectively, our findings identify TAMs as a previously unsuspected mechanism by which the metastasis-suppressor RKIP regulates tumor invasiveness, and further suggest that TNBC patients with decreased RKIP activity and increased TAM infiltration may respond to macrophage-based therapeutics.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line, Tumor / transplantation
  • Chemokine CCL5 / biosynthesis
  • Chemokine CCL5 / genetics
  • Chemokine CCL5 / physiology
  • Chemokines / physiology*
  • Chemotaxis*
  • Cyclohexanes / pharmacology
  • Cyclohexanes / therapeutic use
  • Disease-Free Survival
  • Female
  • Gene Expression Profiling
  • Gene Knockdown Techniques
  • HMGA2 Protein / physiology
  • Heterografts / immunology
  • Humans
  • Macrophages / immunology*
  • Mammary Neoplasms, Experimental / drug therapy
  • Mammary Neoplasms, Experimental / immunology*
  • Maraviroc
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Mice, Nude
  • Neoplasm Metastasis / immunology*
  • Neoplasm Proteins / biosynthesis
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / physiology*
  • Phosphatidylethanolamine Binding Protein / physiology*
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / genetics
  • RNA, Neoplasm / biosynthesis
  • RNA, Neoplasm / genetics
  • Receptors, CCR5 / drug effects
  • Sequence Analysis, RNA
  • Triazoles / pharmacology
  • Triazoles / therapeutic use
  • Triple Negative Breast Neoplasms / immunology*
  • Triple Negative Breast Neoplasms / mortality
  • Tumor Microenvironment / immunology*


  • CCL5 protein, human
  • CCR5 protein, mouse
  • Ccl5 protein, mouse
  • Chemokine CCL5
  • Chemokines
  • Cyclohexanes
  • HMGA2 Protein
  • Neoplasm Proteins
  • PEBP1 protein, human
  • Phosphatidylethanolamine Binding Protein
  • RNA, Messenger
  • RNA, Neoplasm
  • Raf kinase inhibitory protein, mouse
  • Receptors, CCR5
  • Triazoles
  • Maraviroc