Abstract 4986: APG101 inhibits CD95-Ligand induced invasion of glioblastoma in 3D-models <i>in vitro</i> and <i>ex vivo</i>
Authors: Christian Merz, Alexander Strecker, Jaromir Sykora, Oliver Hill, Gieffers Christian, Harald Fricke, Peter Angel, Heike Peterziel
Journal: Cancer Research (2014)

Abstract CD95 (APO-1/Fas) is a receptor that belongs to the Tumor Necrosis Factor Receptor Super Family (TNFRSF). Originally discovered as a death receptor inducing apoptosis upon binding to its ligand CD178 (CD95L/FasL), recent research indicates that CD95 also mediates non-apoptotic functions (e.g. liver regeneration, neuronal development, inflammation and cellular migration/invasion of tumor cells) depending on the cellular context. The potential impact on multiple cellular processes classifies the CD95 pathway as an attractive target for pharmacological interference. Apogenix developed APG101, a recombinant human fusion protein consisting of the extracellular domain of CD95 and the Fc-domain of an IgG antibody for the treatment of glioblastoma, a malignancy characterized by aggressive and invasive growth. Preclinical development activities to elucidate the mode of action of APG101 included the comparison of 3-dimensional in vitro and ex vivo models employing U87-MG and U373-MG cells on murine organotypic brain tissue cultures. Expression of CD95L and CD95 in these cells was reduced by semi-stable transfection of shRNA vectors to assess contribution of the CD95/CD95L system to invasion and proliferation of glioma cell lines and the anti-invasive effect of APG101 treatment. While knockdown of CD95 leads to death of U373-MG cells, we observed significant reduction of proliferation rates in U87-MG cells. The effect of CD95L knockdown on proliferation was only minor in U87-MG and was absent in U373-MG cells. Consistently, we found that treatment of glioma cells with APG101 had no effect on proliferation rates or cell cycle regulation of either cell line. The main effect of CD95L knockdown was a strongly reduced invasiveness of both cell lines in vitro and ex vivo. Invasiveness could be restored by exogenous addition of recombinant CD95L, and this induced invasion was inhibited in the presence of APG101. In contrast, a CD95L binding defective CD95-Fc mutant APG122 did not have an anti-invasive effect in our experimental models. We conclude that: Results obtained from the in vitro and ex vivo assay models reflect each other and are suitable to study the role of CD95 signaling in tumor cell invasion activation of CD95 on glioma cells by CD95L contributes to invasive spread of tumor cells to the surrounding brain tissue, and Apogenix drug APG101 is able to block the pro-invasive activity of CD95L. Based on these observations, an important mode of action for APG101 is inhibition of CD95 ligand induced invasion of glioma cells. Citation Format: Christian Merz, Alexander Strecker, Jaromir Sykora, Oliver Hill, Gieffers Christian, Harald Fricke, Peter Angel, Heike Peterziel. APG101 inhibits CD95-Ligand induced invasion of glioblastoma in 3D-models in vitro and ex vivo. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4986. doi:10.1158/1538-7445.AM2014-4986