184AA3: a xenograft model of ER+ breast adenocarcinoma
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Despite the prevalence and significant morbidity resulting from estrogen receptor positive (ER+) breast adenocarcinomas, there are only a few models of this cancer subtype available for drug development and arguably none for studying etiology. Those models that do exist have questionable clinical relevance. Given our goal of developing luminal models, we focused on six cell lines derived by minimal mutagenesis from normal human breast cells, and asked if any could generate clinically relevant xenografts, which we then extensively characterized. Xenografts of one cell line, 184AA3, consistently formed ER+ adenocarcinomas that had a high proliferative rate and other features consistent with “luminal B” intrinsic subtype. Squamous and spindle cell/mesenchymal differentiation was absent, in stark contrast to other cell lines that we examined or others have reported. We explored intratumoral heterogeneity produced by 184AA3 by immunophenotyping xenograft tumors and cultured cells, and characterized marker expression by immunofluorescence and flow cytometry. A CD44High subpopulation was discovered, yet their tumor forming ability was far less than CD44Low cells. Single cell cloning revealed the phenotypic plasticity of 184AA3, consistent with the intratumoral heterogeneity observed in xenografts. Characterization of ER expression in cultures revealed ER protein and signaling is intact, yet when estrogen was depleted in culture, and in vivo, it did not impact cell or tumor growth, analogous to therapeutically resistant ER+ cancers. This model is appropriate for studies of the etiology of ovarian hormone independent adenocarcinomas, for identification of therapeutic targets, predictive testing, and drug development.
KeywordsLuminal breast cancer models Xenograft Intratumoral heterogeneity Microenvironment
For their invaluable technical assistance during this project, we thank Maria Rojec, Dinah Groesser, Alvin Lo, Sun-Young Lee, Xuefei Tian, and Eva Lee (Lawrence Berkeley National Laboratory). We appreciate also the expertise and help given by Judith Walls and Ed Hubbard (University of California, Davis Center for Comparative Medicine). We express special gratitude also to Michelle Scott of the LBNL flow cytometry and Advanced microscopy facility for her expert technical advice and assistance. Grant support: Innovator award to M.J.B. from the U.S. Department of Defense (W81XWH0810736 and W81XWH12M9532) and in part by National Cancer Institute awards (R37CA064786, R01CA140663, U54CA112970) and by grants from the U.S. Department of Energy, Office of Biological and Environmental Research and Low Dose Scientific Focus Area (Contract No. DE-AC02-05CH1123) and the Breast Cancer Research Foundation. The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Compliance with ethical standards
All procedures performed in studies involving animals were in accordance with the ethical standards of the Lawrence Berkeley National Laboratory, at which all such studies were conducted.
Conflicts of interest
The authors declare that they have no conflict of interest.
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