SCOPE OF WORK Despite advances in therapy, breast cancer remains the second leading cause of cancer related deaths in women. As breast cancer cells are genetically unstable and heterogeneous, focusing on the tumor microenvironment (TME) has the potential to identify new therapeutic targets. One important cytokine in the TME for many cancers is the TGF-β1 ligand that binds TGF-β receptors to promote Smad signaling and gene expression. The TGF-β signaling pathway has well established roles in regulating cellular homeostasis, including proliferation, differentiation and apoptosis. During cancer initiation and progression, the TGF-β signaling pathway is disrupted in a cell and context specific manner. In breast cancer, TGF-β suppresses tumor initiation, but in established cancers, promotes cancer progression to increase invasion and metastasis. As such, TGF-β is an attractive cancer therapeutic target. However, targeting the TGF-β signaling pathway directly has not been successful in the clinic partially due to an incomplete understanding of how TGF-β’s role changes during cancer progression. To gain insight into factors that may regulate TGF-β signaling in the tumor microenvironment, we performed a proteomic screen of the breast cancer secretome and identified the integrin binding protein βIGH3 as a protein able to promote TGF-β signaling. CRISPR/Cas9 silencing of βIGH3 decreases tumor growth and metastasis in a breast cancer model. This decrease in tumor progression was also associated with decreased TGF-β signaling in vivo. We hypothesize that βIGH3 increases breast cancer progression by promoting TGF-β-induced EMT, migration and invasion, with effects on TGF-β signaling mediated by increasing active TGF-β1 ligand through proteases. To address this hypothesis, we propose two aims. Aim 1: To establish whether βIGH3 induces TGF-β signaling by increasing the activation of mature TGF-β1 ligand from the latent-TGFβ precursor. Aim 2: To determine whether βIGH3 increases metastasis by promoting TGF-β signaling and TGF-β induced EMT associated migration, invasion and extravasation. Defining the detailed mechanism of how βIGH3 increases tumorigenesis through TGF-β signaling could potentially lead to a new therapeutic target in breast cancer.

PROJECT NARRATIVE Breast cancer is the second leading cause of cancer related deaths for women in the US, with 1 in 8 women developing breast cancer in their life time. While there is an excellent outcome for most breast cancer patients with localized and hormone responsive breast cancer, those with metastatic disease tend to do poorly due to limited treatment options; therefore, it is important to identify novel therapeutic targets in advanced breast cancer. The goal for our research is to determine how the extracellular matrix protein transforming growth factor beta induced (TGFBI or βIGH3) is capable of promoting metastasis, and how we might target this secreted protein as a potential cancer therapy for these patients.