Spontaneous dormancy of metastatic breast cancer cells in an all human liver microphysiologic system

Background:Metastatic outgrowth in breast cancer can occur years after a seeming cure. Existing model systems of dormancy are limited as they do not recapitulate human metastatic dormancy without exogenous manipulations and are unable to query early events of micrometastases.Methods:Here, we describe a human ex vivo hepatic microphysiologic system. The system is established with fresh human hepatocytes and non-parenchymal cells (NPCs) creating a microenvironment into which breast cancer cells (MCF7 and MDA-MB-231) are added.Results:The hepatic tissue maintains function through 15 days as verified by liver-specific protein production and drug metabolism assays. The NPCs form an integral part of the hepatic niche, demonstrated within the system through their participation in differential signalling cascades and cancer cell outcomes. Breast cancer cells intercalate into the hepatic niche without interfering with hepatocyte function. Examination of cancer cells demonstrated that a significant subset enter a quiescent state of dormancy as shown by lack of cell cycling (EdU− or Ki67−). The presence of NPCs altered the cancer cell fraction entering quiescence, and lead to differential cytokine profiles in the microenvironment effluent.Conclusions:These findings establish the liver microphysiologic system as a relevant model for the study of breast cancer metastases and entry into dormancy.

[1]  C. Hughes,et al.  Of Mice and Not Men: Differences between Mouse and Human Immunology , 2004, The Journal of Immunology.

[2]  R. Eils,et al.  Systemic spread is an early step in breast cancer. , 2008, Cancer cell.

[3]  Daniel A Fletcher,et al.  Tissue Geometry Determines Sites of Mammary Branching Morphogenesis in Organotypic Cultures , 2006, Science.

[4]  David J Beebe,et al.  Hedgehog signaling in myofibroblasts directly promotes prostate tumor cell growth. , 2012, Integrative biology : quantitative biosciences from nano to macro.

[5]  J. Brugge,et al.  Use of Three-Dimensional Basement Membrane Cultures to Model Oncogene-Induced Changes in Mammary Epithelial Morphogenesis , 2004, Journal of Mammary Gland Biology and Neoplasia.

[6]  Celeste M Nelson,et al.  Tissue geometry patterns epithelial–mesenchymal transition via intercellular mechanotransduction , 2010, Journal of cellular biochemistry.

[7]  I. Macdonald,et al.  Temporal progression of metastasis in lung: cell survival, dormancy, and location dependence of metastatic inefficiency. , 2000, Cancer research.

[8]  Alan Wells,et al.  Partial Mesenchymal to Epithelial Reverting Transition in Breast and Prostate Cancer Metastases , 2012, Cancer Microenvironment.

[9]  B. E. F. Isher,et al.  Twenty-year follow-up of a randomized trial comparing total mastectomy, lumpectomy, and lumpectomy plus irradiation for the treatment of invasive breast cancer. , 2002 .

[10]  Alan Wells,et al.  Novel three-dimensional organotypic liver bioreactor to directly visualize early events in metastatic progression. , 2007, Advances in cancer research.

[11]  K. Luzzi,et al.  Multistep nature of metastatic inefficiency: dormancy of solitary cells after successful extravasation and limited survival of early micrometastases. , 1998, The American journal of pathology.

[12]  B. Weigelt,et al.  The need for complex 3D culture models to unravel novel pathways and identify accurate biomarkers in breast cancer. , 2014, Advanced drug delivery reviews.

[13]  Mina J. Bissell,et al.  The perivascular niche regulates breast tumor dormancy , 2013, Nature Cell Biology.

[14]  K. Pantel,et al.  Clinical relevance and biology of circulating tumor cells , 2011, Breast Cancer Research.

[15]  S. Friedman,et al.  Proangiogenic role of tumor‐activated hepatic stellate cells in experimental melanoma metastasis , 2003, Hepatology.

[16]  Alan Wells,et al.  The dormancy dilemma: quiescence versus balanced proliferation. , 2013, Cancer research.

[17]  Claudia Fischbach,et al.  Microfluidic culture models of tumor angiogenesis. , 2010, Tissue engineering. Part A.

[18]  J. Izbicki,et al.  Differential expression of proliferation-associated molecules in individual micrometastatic carcinoma cells. , 1993, Journal of the National Cancer Institute.

[19]  A. Børresen-Dale,et al.  Re-expression of E-cadherin, alpha-catenin and beta-catenin, but not of gamma-catenin, in metastatic tissue from breast cancer patients [seecomments]. , 2000, The Journal of pathology.

[20]  I. Fidler,et al.  Metastasis: Quantitative Analysis of Distribution and Fate of Tumor Emboli Labeled With 125I-5-Iodo-2′ -deoxyuridine , 1970 .

[21]  Alan Wells,et al.  Breast carcinoma cells re-express E-cadherin during mesenchymal to epithelial reverting transition , 2010, Molecular Cancer.

[22]  Andrej J. Savol,et al.  Modeling Boundary Conditions for Balanced Proliferation in Metastatic Latency , 2013, Clinical Cancer Research.

[23]  L. Griffith,et al.  A microfabricated array bioreactor for perfused 3D liver culture. , 2002, Biotechnology and bioengineering.

[24]  I. Bross,et al.  Am Autopsy Study of Some Routes of Dissemination of Cancer of the Breast , 1973, British Journal of Cancer.

[25]  P. So,et al.  The FASEB Journal • Research Communication , 2007 .

[26]  R. Demicheli,et al.  Breast cancer recurrence dynamics following adjuvant CMF is consistent with tumor dormancy and mastectomy-driven acceleration of the metastatic process. , 2005, Annals of oncology : official journal of the European Society for Medical Oncology.

[27]  G. Hortobagyi,et al.  Overview of resistance to systemic therapy in patients with breast cancer. , 2007, Advances in experimental medicine and biology.

[28]  J. Holland,et al.  Vinblastine, adriamycin, thiotepa, and halotestin (VATH). Therapy for advanced breast cancer refractory to prior chemotherapy , 1978, Cancer.

[29]  I. N. Crispe,et al.  The liver as a lymphoid organ. , 2009, Annual review of immunology.

[30]  B. Davidson,et al.  A prospective study of isolated human hepatocyte function following liver resection for colorectal liver metastases: the effects of prior exposure to chemotherapy. , 2006, Journal of hepatology.

[31]  D. Redelmeier,et al.  Translation of research evidence from animals to humans. , 2006, JAMA.

[32]  Gerhard Christofori,et al.  Mouse models of breast cancer metastasis , 2006, Breast Cancer Research.

[33]  A. Wells,et al.  Mesenchymal–epithelial transition (MET) as a mechanism for metastatic colonisation in breast cancer , 2012, Cancer and Metastasis Reviews.

[34]  M. Rubin,et al.  E-cadherin expression in primary carcinomas of the breast and its distant metastases , 2003, Breast Cancer Research.

[35]  H. Ford,et al.  Epithelial-Mesenchymal Transition in Cancer: Parallels Between Normal Development and Tumor Progression , 2010, Journal of Mammary Gland Biology and Neoplasia.

[36]  Paul A. Khavari,et al.  Invasive 3-Dimensional Organotypic Neoplasia from Multiple Normal Human Epithelia , 2010, Nature Medicine.

[37]  Alan Wells,et al.  All-human microphysical model of metastasis therapy , 2013, Stem Cell Research & Therapy.

[38]  S. Caritis,et al.  A sensitive and specific CYP cocktail assay for the simultaneous assessment of human cytochrome P450 activities in primary cultures of human hepatocytes using LC-MS/MS. , 2013, Journal of pharmaceutical and biomedical analysis.

[39]  B. Teicher.  Tumor models for efficacy determination , 2006, Molecular Cancer Therapeutics.

[40]  A. Børresen-Dale,et al.  Re‐expression of E‐cadherin, α‐catenin and β‐catenin, but not of γ‐catenin, in metastatic tissue from breast cancer patients , 2000 .

[41]  A. Dixit,et al.  ERRβ signalling through FST and BCAS2 inhibits cellular proliferation in breast cancer cells , 2014, British Journal of Cancer.

[42]  M. Junttila,et al.  Influence of tumour micro-environment heterogeneity on therapeutic response , 2013, Nature.

[43]  A. Tutt,et al.  A novel model of dormancy for bone metastatic breast cancer cells. , 2013, Cancer research.

[44]  K. Hunter,et al.  Modeling metastasis in vivo. , 2004, Carcinogenesis.

[45]  C. Selden,et al.  Engineering the liver , 2002 .

[46]  J. Rautela,et al.  The emerging role of immunosurveillance in dictating metastatic spread in breast cancer. , 2013, Cancer research.