Breast cancer is a leading cause of mortality in women worldwide, in part due to the tumor microenvironment which increases tumor heterogeneity and abets tumor growth. Fibroblasts are cells of mesenchymal origin that are an important component of normal and tumor stroma. Genetic alterations in these cells were shown by several groups including our own to cause fibroblast activation and fuel tumor progression giving rise to more aggressive disease. Platelet-Derived Growth Factor Receptor (PDGFR) alpha is a receptor tyrosine kinase that is chiefly expressed in mesenchymal cells such as fibroblasts. Ligand binding (PDGFAA) activates this receptor. PDGFRα signaling plays critical roles in development and aberrant signaling is seen in several types of cancer, such as lung, pancreas, GI and brain. The central goal of this study was to elucidate the role of stromal PDGFRα in breast cancer development and metastasis, where its role remains largely unknown. To address this goal, we developed a genetic mouse model of stromal activation of PDGFRα in the mammary gland by crossing an auto-activating Pdgfra mutant allele with a mesenchymal specific Cre recombinase. We found that stromal PDGFRα activation completely abrogated postnatal mammary gland ductal formation, with significantly reduced terminal end bud formation. PDGFRα activation also led to progressive fibrosis in the mouse mammary fat pad. As early as four weeks of age, mammary collagen (trichrome staining; second harmonic generation) and hyaluronan deposition (Alcian Blue) was greatly increased in vivo. In fact, this increase in collagen and hyaluronan deposition in mutant animals is believed to be responsible for the observed increased in stiffness of mutant mammary tissue (atomic force microscopy {AFM}). pFAK, which can be activated due to mechanical stress, was increased in mammary epithelia of the mutant mice in vivo corroborating the AFM results. Further, when tumor cells were injected into the mammary glands of the PDGFRα mutants, tumors grew faster as compared to controls. Importantly, we found that mRNA expression of PDGFRA correlates with worsened patient outcomes in HER2+ disease, while expression of both the ligand (PDGFA) and the receptor were found to correlate with increased incidence of lung metastases. We further discovered that in HER2+ patients, PDGFRA levels correlate with breast density. Breast density is the third strongest risk factor for breast cancer, and is directly related to collagen deposition and breast stiffness, thus suggesting a novel predictive role of PDGFRA as a molecular readout of stiffness and density. Studies are underway to utilize mouse models of HER2+ breast cancer to study both primary tumor growth and metastases. Taken together, our mouse studies and paralleling human data analyses suggest that the stromal PDGFRα signaling provides a novel theranostic window in breast cancer treatment and prognosis.

Citation Format: Anisha Mathur Hammer, Gina M. Sizemore, Vasudha Shukla, Steven T. Sizemore, Maria Cuitino, Cynthia J. Timmers, Quinn Verfurth, Arnab Chakravarti, Gustavo W. Leone, Samir N. Ghadiali, Michael C. Ostrowski. PDGFR-α induced stiffness abrogates mammary ductal development and enhances tumorigenesis in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4337. doi:10.1158/1538-7445.AM2017-4337