Abstract B79: TIMP-2 mediated growth inhibition of human glioma U87MG is accompanied by elevated IGFBP7 and FSP-1/S100A4 expression at the tumor/stroma interface

Abstract

Abstract The tissue inhibitor of matrix metalloproteinase–2 (TIMP-2) belongs to a small family of multifunctional secreted proteins that inhibit matrix metalloproteases (MMPs), proteases principally associated with promoting tumor invasion and metastasis. We have previously shown that forced TIMP-2 expression inhibits human lung tumor growth through MMP-independent mechanisms including tumor cell apoptosis and upregulation of insulin growth factor binding protein-7 (IGFPB7), a protein reported to function as a potential tumor suppressor in a variety of tumor types including human glioma. In the current study, we used immunohistochemistry, TUNEL assay, and dual immunofluorescence to assess the growth inhibitory effects of TIMP-2 in human glioma xenografts generated using U87MG cells lentivirally transfected to overexpress the human TIMP-2 cDNA sequence (U87MG-T2) or vector alone (U87MG-EV). Compared to EV controls, U87MG-T2 xenografts demonstrated a significant inhibition of tumor growth, accompanied by increased tumor cell apoptosis at the tumor edge. Moreover, a dramatic increase in IGFBP7 and fibroblast specific protein-1 (FSP1)/S100A4 was detected in distinct subpopulations of non-tumor cells at the tumor/stroma interface, whereas both markers were sparsely observed in U87MG-EV controls. In an effort to determine the origin of IGFBP7 positive cells, we performed dual immunofluorescence using antibodies to the blood vessel marker CD34, FSP-1/S100A4, and human mitochondria (clone 113-1). IGFBP7 positive cells did not co-express any of the three markers analyzed, indicating that IGFBP7 positive cells are host derived and consist of cells other than blood vessels or stromal fibroblasts. We conclude that endogenous TIMP-2 significantly inhibits human glioma tumor growth in vivo through tumor cell apoptosis and tumor-host interactions involving discrete subsets of non-tumor cells in the adjacent tumor microenvironment. Citation Format: Sandra Jensen-Taubman, Dimitra Bourboulia, Huiying Han, Laurie Shuman Moss, Beiyang Wei, William G. Stetler-Stevenson. TIMP-2 mediated growth inhibition of human glioma U87MG is accompanied by elevated IGFBP7 and FSP-1/S100A4 expression at the tumor/stroma interface. [abstract]. In: Abstracts: AACR Special Conference on Cellular Heterogeneity in the Tumor Microenvironment; 2014 Feb 26-Mar 1; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2015;75(1 Suppl):Abstract nr B79. doi:10.1158/1538-7445.CHTME14-B79