Abstract B045: uPAR-targeted theranostic nanoparticles effectively decrease expression of IGF-1R and Ki67 in drug-resistant triple-negative breast cancer human xenograft
Authors: Miller-Kleinhenz Jasmine M. , Zhou Hongyu , Qian Weiping , O'Regan Ruth , Zelnak Amelia , Styblo Toncred , Yang Lily 
Published: May 2015

Background: Triple-negative breast cancer (TNBC) is the most aggressive breast cancer subtype that affects about 20% of breast cancer patients and has a high incidence in young African American women. Currently the only therapeutic options for TNBC patients is chemotherapy, however, more than 60% of TNBC patients are highly resistant to chemotherapeutic treatment. The development of novel therapeutic approaches is essential to enable treatment of patients with chemo-resistant TNBC.

Our lab focuses on the development of receptor-targeted theranostic nanoparticles. We have developed a class of urokinase plasminogen activator receptor (uPAR) targeted theranostic magnetic iron oxide nanoparticles (IONPs) carrying therapeutic agents that are clinically beneficial to TNBC patients, including DNA damaging agents, such as doxorubicin (Dox) or cisplatin (Cis). We are targeting uPAR because it is highly expressed in malignant cancers including aggressive breast cancer tissues and tumor stromal cells that are enriched in TNBC tissues.

Methods: Tumor fragments from TNBC patients, obtained from a Phase II clinical trial that is ongoing at the Breast Cancer Clinic in the Winship Cancer Institute, were implanted in the mammary fat pad of SCID mice. The tumor xenografts grew to about 1 cm in diameter in 12 to 15 weeks. Tumor fragments were then implanted into the mammary fat pad and passaged through nude mice for testing the therapeutic efficacy of the uPAR-targeted nanoparticle-drugs. Histological analysis of post-chemotherapy TNBC and primary tumor xenograft tissues with or without uPAR-targeted IONP-Dox was conducted using immunohistochemistry and dual immunofluorescence techniques.

Results: We have previously reported that the uPAR-targeted IONP-Dox causes tumor growth inhibition following systemic delivery. Using CD44 and uPAR as biomarkers we also reported histological analysis of post-chemotherapy TNBC and primary tumor xenograft tissues showed high levels of CD44+/CD24- tumor cells and up-regulation of uPAR. Primary TNBC xenografts treated with conventional Dox had increased levels of CD44+/CD24- cells and strong uPAR expression. Conversely, the tumor tissues from the uPAR-targeted IONP-Dox treated group showed decreased levels of CD44+/CD24- and uPAR positive cells.

We have further investigated the biomarkers insulin-like growth factor 1 receptor (IGF-1R) and Ki67. IGF-1R has been shown to be overexpressed in more than 50% of TNBCs and may be associated with the drug-resistant TNBC phenotype, while Ki67 is a conventional biomarker for proliferation. We observed that uPAR-targeted IONP- Dox treatment decreased IGF-1R expression compared to the control group and the group treated with conventional Dox. We also observed that Ki67 expression was decreased in the uPAR-targeted IONP- Dox treatment group compared to control group and the group treated with conventional Dox.

Conclusions: Results of our study demonstrated that uPAR-targeted theranostic IONPs selectively delivers therapeutic payloads of the chemotherapeutic drug Dox to inhibit the growth of uPAR positive tumor cells. It is likely that uPAR expressing tumor cells are an aggressive and potentially drug resistant cell population that overexpress breast cancer stem-like cell biomarker (CD44+/CD24-) as well as up-regulation of a growth factor receptor, IGF-1R. These results support further investigation of theranostic nanoparticles as an approach to overcome drug resistance in TNBC.

Citation Format: Jasmine M. Miller-Kleinhenz, Hongyu Zhou, Weiping Qian, Ruth O'Regan, Amelia Zelnak, Toncred Styblo, Lily Yang. uPAR-targeted theranostic nanoparticles effectively decrease expression of IGF-1R and Ki67 in drug-resistant triple-negative breast cancer human xenograft. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Breast Cancer Research: Genetics, Biology, and Clinical Applications; Oct 3-6, 2013; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2013;11(10 Suppl):Abstract nr B045.