Tumor-associated macrophages drive spheroid formation during early transcoelomic metastasis of ovarian cancer.

Tumor-associated macrophages (TAMs) can influence ovarian cancer growth, migration, and metastasis, but the detailed mechanisms underlying ovarian cancer metastasis remain unclear. Here, we have shown a strong correlation between TAM-associated spheroids and the clinical pathology of ovarian cancer. Further, we have determined that TAMs promote spheroid formation and tumor growth at early stages of transcoelomic metastasis in an established mouse model for epithelial ovarian cancer. M2 macrophage-like TAMs were localized in the center of spheroids and secreted EGF, which upregulated αMβ2 integrin on TAMs and ICAM-1 on tumor cells to promote association between tumor cells and TAM. Moreover, EGF secreted by TAMs activated EGFR on tumor cells, which in turn upregulated VEGF/VEGFR signaling in surrounding tumor cells to support tumor cell proliferation and migration. Pharmacological blockade of EGFR or antibody neutralization of ICAM-1 in TAMs blunted spheroid formation and ovarian cancer progression in mouse models. These findings suggest that EGF secreted from TAMs plays a critical role in promoting early transcoelomic metastasis of ovarian cancer. As transcoelomic metastasis is also associated with many other cancers, such as pancreatic and colon cancers, our findings uncover a mechanism for TAM-mediated spheroid formation and provide a potential target for the treatment of ovarian cancer and other transcoelomic metastatic cancers.

[1]  G. Freeman,et al.  Therapeutic PD-1 pathway blockade augments with other modalities of immunotherapy T-cell function to prevent immune decline in ovarian cancer. , 2013, Cancer research.

[2]  M. Ewen,et al.  CCL18 from tumor-associated macrophages promotes breast cancer metastasis via PITPNM3. , 2011, Cancer cell.

[3]  S. Rafii,et al.  VEGFR1-positive haematopoietic bone marrow progenitors initiate the pre-metastatic niche , 2005, Nature.

[4]  J. McGee,et al.  Intact LKB1 activity is required for survival of dormant ovarian cancer spheroids , 2015, Oncotarget.

[5]  R. Agarwal,et al.  Mechanisms of transcoelomic metastasis in ovarian cancer. , 2006, The Lancet. Oncology.

[6]  S. Gordon,et al.  Ovarian Cancer Cells Polarize Macrophages Toward A Tumor-Associated Phenotype1 , 2006, The Journal of Immunology.

[7]  A. Jemal,et al.  Cancer Statistics, 2009 , 2009, CA: a cancer journal for clinicians.

[8]  L. Twiggs,et al.  Twenty-five year outcome of sequential abdominal radiotherapy and melphalan:implications for future management of epithelial carcinoma of the ovary. , 2005, Gynecologic oncology.

[9]  S. Fagerholm,et al.  alpha-Chain phosphorylation of the human leukocyte CD11b/CD18 (Mac-1) integrin is pivotal for integrin activation to bind ICAMs and leukocyte extravasation. , 2006, Blood.

[10]  D. Hanahan,et al.  Hallmarks of Cancer: The Next Generation , 2011, Cell.

[11]  J. Au,et al.  Intraperitoneal therapy for peritoneal cancer. , 2010, Future oncology.

[12]  Yingke Xu,et al.  A network of interactions enables CCM3 and STK24 to coordinate UNC13D-driven vesicle exocytosis in neutrophils. , 2013, Developmental cell.

[13]  Xia Li,et al.  Over‐expression of LAPTM4B is associated with poor prognosis and chemotherapy resistance in stages III and IV epithelial ovarian cancer , 2011, Journal of surgical oncology.

[14]  G. Coukos,et al.  Deciphering and reversing tumor immune suppression. , 2013, Immunity.

[15]  G. Cline,et al.  Functional polarization of tumour-associated macrophages by tumour-derived lactic acid , 2014, Nature.

[16]  M. Zucchetti,et al.  Role of macrophage targeting in the antitumor activity of trabectedin. , 2013, Cancer cell.

[17]  E. Lengyel.  Ovarian cancer development and metastasis. , 2010, The American journal of pathology.

[18]  A. Jimeno,et al.  Randomized phase III study of erlotinib versus observation in patients with no evidence of disease progression after first-line platin-based chemotherapy for ovarian carcinoma: a European Organisation for Research and Treatment of Cancer-Gynaecological Cancer Group, and Gynecologic Cancer Intergroup , 2014, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[19]  N. Teng,et al.  FACS analysis of peritoneal lymphocytes in ovarian cancer and control patients. , 1994, Immunobiology.

[20]  N. Weiss,et al.  Predictive value of symptoms for early detection of ovarian cancer. , 2010, Journal of the National Cancer Institute.

[21]  John Condeelis,et al.  Macrophages: Obligate Partners for Tumor Cell Migration, Invasion, and Metastasis , 2006, Cell.

[22]  E. Yordan,et al.  Long-term follow-up and prognostic factor analysis in advanced ovarian carcinoma: the Gynecologic Oncology Group experience. , 1991, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[23]  Y. Suárez,et al.  Cutting Edge: TNF-Induced MicroRNAs Regulate TNF-Induced Expression of E-Selectin and Intercellular Adhesion Molecule-1 on Human Endothelial Cells: Feedback Control of Inflammation , 2009, The Journal of Immunology.

[24]  A. Beaudet,et al.  Role of the intercellular adhesion molecule-1(ICAM-1) in endotoxin-induced pneumonia evaluated using ICAM-1 antisense oligonucleotides, anti-ICAM-1 monoclonal antibodies, and ICAM-1 mutant mice. , 1996, The Journal of clinical investigation.

[25]  W. Min,et al.  MyD88-dependent, superoxide-initiated inflammation is necessary for flow-mediated inward remodeling of conduit arteries , 2008, The Journal of experimental medicine.

[26]  R. Drapkin,et al.  Combined therapy with thrombospondin‐1 type I repeats (3TSR) and chemotherapy induces regression and significandy improves survival in a preclinical model of advanced stage epithelial ovarian cancer , 2015, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[27]  P. Carmeliet,et al.  HRG inhibits tumor growth and metastasis by inducing macrophage polarization and vessel normalization through downregulation of PlGF. , 2011, Cancer cell.

[28]  E. Lengyel,et al.  Expression of the homeobox gene HOXA9 in ovarian cancer induces peritoneal macrophages to acquire an M2 tumor-promoting phenotype. , 2014, The American journal of pathology.

[29]  P. Biberfeld,et al.  MALA‐2, mouse homologue of human adhesion molecule ICAM‐1 (CD54) , 1989, European journal of immunology.

[30]  C. Lewis,et al.  Macrophage regulation of tumor responses to anticancer therapies. , 2013, Cancer cell.

[31]  Jeffrey W. Pollard,et al.  Macrophage Diversity Enhances Tumor Progression and Metastasis , 2010, Cell.

[32]  G. Mor,et al.  Epithelial ovarian cancer cells secrete functional Fas ligand. , 2003, Cancer research.

[33]  E. Puré,et al.  Reduced contact sensitivity reactions in mice treated with monoclonal antibodies to leukocyte function-associated molecule-1 and intercellular adhesion molecule-1. , 1993, Journal of immunology.

[34]  S. Meri,et al.  Ascitic complement system in ovarian cancer , 2005, British Journal of Cancer.

[35]  R. Weinberg,et al.  The tumour-induced systemic environment as a critical regulator of cancer progression and metastasis , 2014, Nature Cell Biology.

[36]  S. Rafii,et al.  Inflammation joins the "niche". , 2008, Cancer cell.

[37]  Will Liao,et al.  The cellular and molecular origin of tumor-associated macrophages , 2014, Science.

[38]  Ming-zhu Yin,et al.  LAPTM4B overexpression is a novel predictor of epithelial ovarian carcinoma metastasis , 2011, International journal of cancer.

[39]  Kathleen R. Cho,et al.  Myeloid-derived suppressor cells enhance stemness of cancer cells by inducing microRNA101 and suppressing the corepressor CtBP2. , 2013, Immunity.

[40]  A. Jemal,et al.  Cancer statistics, 2012 , 2012, CA: a cancer journal for clinicians.

[41]  J. Massagué,et al.  Macrophage binding to receptor VCAM-1 transmits survival signals in breast cancer cells that invade the lungs. , 2011, Cancer cell.

[42]  R. Jain,et al.  Polarization of tumor-associated macrophages: a novel strategy for vascular normalization and antitumor immunity. , 2011, Cancer cell.

[43]  A. Skubitz,et al.  Ovarian carcinoma ascites spheroids adhere to extracellular matrix components and mesothelial cell monolayers. , 2004, Gynecologic oncology.

[44]  Andrew Burgess,et al.  Loss of human Greatwall results in G2 arrest and multiple mitotic defects due to deregulation of the cyclin B-Cdc2/PP2A balance , 2010, Proceedings of the National Academy of Sciences.

[45]  A. Reuss,et al.  Bevacizumab combined with chemotherapy for platinum-resistant recurrent ovarian cancer: The AURELIA open-label randomized phase III trial. , 2014, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[46]  S. Cherry,et al.  Infection-induced type I interferons activate CD11b on B-1 cells for subsequent lymph node accumulation , 2015, Nature Communications.

[47]  Prahlad T. Ram,et al.  Hematogenous metastasis of ovarian cancer: rethinking mode of spread. , 2014, Cancer cell.

[48]  E. Ratner,et al.  Solitomab, an epithelial cell adhesion molecule/CD3 bispecific antibody (BiTE), is highly active against primary chemotherapy‐resistant ovarian cancer cell lines in vitro and fresh tumor cells ex vivo , 2015, Cancer.

[49]  Erik Sahai,et al.  Macrophages promote the invasion of breast carcinoma cells via a colony-stimulating factor-1/epidermal growth factor paracrine loop. , 2005, Cancer research.

[50]  N. Van Rooijen,et al.  Macrophages mediate inflammation-enhanced metastasis of ovarian tumors in mice. , 2007, Cancer research.

[51]  A. Skubitz,et al.  Beta 1-integrins regulate the formation and adhesion of ovarian carcinoma multicellular spheroids. , 2001, The American journal of pathology.

[52]  R. Freedman,et al.  Defective Antitumor Function of Monocyte-Derived Macrophages from Epithelial Ovarian Cancer Patients , 2006, Clinical Cancer Research.

[53]  G. Rice,et al.  α2β1 integrin affects metastatic potential of ovarian carcinoma spheroids by supporting disaggregation and proteolysis , 2007, Journal of Carcinogenesis.

[54]  Jinghang Zhang,et al.  CCL2 recruits inflammatory monocytes to facilitate breast tumor metastasis , 2011, Nature.

[55]  J. Shih,et al.  The VEGF-C/Flt-4 axis promotes invasion and metastasis of cancer cells. , 2006, Cancer cell.

[56]  N. Dubrawsky.  Cancer statistics , 1989, CA: a cancer journal for clinicians.

[57]  F. Takei.  Inhibition of mixed lymphocyte response by a rat monoclonal antibody to a novel murine lymphocyte activation antigen (MALA-2). , 1985, Journal of immunology.

[58]  F. Takei,et al.  Molecular cloning of murine intercellular adhesion molecule (ICAM‐1). , 1989, The EMBO journal.

[59]  J. McGee,et al.  TGFβ signaling regulates epithelial-mesenchymal plasticity in ovarian cancer ascites-derived spheroids. , 2016, Endocrine-related cancer.

[60]  David A. Cheresh,et al.  Integrins in cancer: biological implications and therapeutic opportunities , 2010, Nature Reviews Cancer.