Abstract A46: Hyaluronan-dependent growth of human triple negative breast cancer MDA-MB-468 in a mouse xenograft model with HA-high stroma
Authors: Zhao Chunmei , Marella Mathieu , Huang Lei , Kultti Anne , Zimmerman Susan , Chou Caroline EN , Bahn Jesse , Radi Adrian , Huang Zhongdong , Shepard H Michael , Thanos Christopher D. 
Published: October 2016

Hyaluronan (also known as hyaluronic acid or HA) is a megadalton sized glycosaminoglycan present in the extracellular matrix. The abnormal accumulation of HA in the solid tumor microenvironment (TME) of several human cancers, including pancreatic, breast, colon and prostate, has been well studied and is associated with poor prognosis. In certain breast cancer specimens with a HA-high status, HA is found to be significantly associated with the stromal compartment rather than with tumor cells. To understand the interaction between tumor cells and a HA containing stromal compartment, we engineered a HA-high stromal cell line by lentiviral transduction of the human HAS3 gene into Balb/c 3T3 fibroblast cells. The 3T3/HAS3 cells produced significant amounts of HA in vitro, which was shown to bind to the HA-low/CD44-high breast cancer cell line MDA-MB-468 at the cell surface. When co-grafted with MDA-MB-468 cells in mice, 3T3/HAS3 cells promoted the in vivo growth of MDA-MB-468 cells. Immunohistochemical analysis of tumor xenograft samples showed that MDA-MB-468 cells were surrounded by HA-high stromal cells, closely resembling the tumor morphology observed in human breast cancer specimens. Tumor growth in this co-grafting model was highly dependent on the presence of 3T3/HAS3 cells, and required HA produced by 3T3/HAS3 cells. Ganciclovir blocked the growth of MDA-MB-468 co-inoculated with 3T3/HAS3/hsv-TK, as did HA removal either by the expression of the HA degrading enzyme PH20, directly in 3T3/HAS3 cells or by exogenous administration of PEGPH20, a pegylated version of PH20 engineered for extended half-life. Interestingly, the presence of 3T3/HAS3 cells was found to be most critical during early stages of tumor progression and was non-essential for tumor maintenance once tumors were fully established, as ganciclovir had little impact on tumor growth when tumor size was larger than 500 mm3 but prevented tumor growth when administered at inoculation and inhibited tumor growth when treatment started at a smaller tumor size (200 mm3, 90.4% TGI). We have developed a tumor xenograft model that mimics breast cancer with a HA-high status in the tumor stromal compartment. Further characterization of this model will provide insight into the mechanisms by which increased levels of HA and the associated changes in the TME promote disease progression in breast cancer.

Citation Format: Chunmei Zhao, Mathieu Marella, Lei Huang, Anne Kultti, Susan Zimmerman, Caroline EN Chou, Jesse Bahn, Adrian Radi, Zhongdong Huang, H Michael Shepard, Christopher D. Thanos. Hyaluronan-dependent growth of human triple negative breast cancer MDA-MB-468 in a mouse xenograft model with HA-high stroma. [abstract]. In: Proceedings of the AACR Special Conference: Function of Tumor Microenvironment in Cancer Progression; 2016 Jan 7–10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2016;76(15 Suppl):Abstract nr A46.