%0 Journal Article %1 Moller2014 %A Möller, Yvonne %A Siegemund, Martin %A Beyes, Sven %A Herr, Ricarda %A Lecis, Daniele %A Delia, Domenico %A Kontermann, Roland %A Brummer, Tilman %A Pfizenmaier, Klaus %A Olayioye, Monilola A. %D 2014 %E Souglakos, John %J PLoS ONE %K 2014 izi kontermann olayioye pfizenmaier %N 9 %P e107165 %R 10.1371/journal.pone.0107165 %T EGFR-targeted TRAIL and a Smac mimetic synergize to overcome apoptosis resistance in KRAS mutant colorectal cancer cells %U http://www.ncbi.nlm.nih.gov/pubmed/25198428 %V 9 %X TRAIL is a death receptor ligand that induces cell death preferentially in tumor cells. Recombinant soluble TRAIL, however, performs poorly as an anti-cancer therapeutic because oligomerization is required for potent biological activity. We previously generated a diabody format of tumor-targeted TRAIL termed Db$\alpha$EGFR-scTRAIL, comprising single-stranded TRAIL molecules (scTRAIL) and the variable domains of a humanized variant of the EGFR blocking antibody Cetuximab. Here we define the bioactivity of Db$\alpha$EGFR-scTRAIL with regard to both EGFR inhibition and TRAIL receptor activation in 3D cultures of Caco-2 colorectal cancer cells, which express wild-type K-Ras. Compared with conventional 2D cultures, Caco-2 cells displayed strongly enhanced sensitivity toward Db$\alpha$EGFR-scTRAIL in these 3D cultures. We show that the antibody moiety of Db$\alpha$EGFR-scTRAIL not only efficiently competed with ligand-induced EGFR function, but also determined the apoptotic response by specifically directing Db$\alpha$EGFR-scTRAIL to EGFR-positive cells. To address how aberrantly activated K-Ras, which leads to Cetuximab resistance, affects Db$\alpha$EGFR-scTRAIL sensitivity, we generated stable Caco-2tet cells inducibly expressing oncogenic K-RasG12V. In the presence of doxycycline, these cells showed increased resistance to Db$\alpha$EGFR-scTRAIL, associated with the elevated expression of the anti-apoptotic proteins cIAP2, Bcl-xL and FlipS. Co-treatment of cells with the Smac mimetic SM83 restored the Db$\alpha$EGFR-scTRAIL-induced apoptotic response. Importantly, this synergy between Db$\alpha$EGFR-scTRAIL and SM83 also translated to 3D cultures of oncogenic K-Ras expressing HCT-116 and LoVo colorectal cancer cells. Our findings thus support the notion that Db$\alpha$EGFR-scTRAIL therapy in combination with apoptosis-sensitizing agents may be promising for the treatment of EGFR-positive colorectal cancers, independently of their KRAS status. %@ 1932-6203 (Electronic)$\backslash$r1932-6203 (Linking)

@article{Moller2014, abstract = {TRAIL is a death receptor ligand that induces cell death preferentially in tumor cells. Recombinant soluble TRAIL, however, performs poorly as an anti-cancer therapeutic because oligomerization is required for potent biological activity. We previously generated a diabody format of tumor-targeted TRAIL termed Db$\alpha$EGFR-scTRAIL, comprising single-stranded TRAIL molecules (scTRAIL) and the variable domains of a humanized variant of the EGFR blocking antibody Cetuximab. Here we define the bioactivity of Db$\alpha$EGFR-scTRAIL with regard to both EGFR inhibition and TRAIL receptor activation in 3D cultures of Caco-2 colorectal cancer cells, which express wild-type K-Ras. Compared with conventional 2D cultures, Caco-2 cells displayed strongly enhanced sensitivity toward Db$\alpha$EGFR-scTRAIL in these 3D cultures. We show that the antibody moiety of Db$\alpha$EGFR-scTRAIL not only efficiently competed with ligand-induced EGFR function, but also determined the apoptotic response by specifically directing Db$\alpha$EGFR-scTRAIL to EGFR-positive cells. To address how aberrantly activated K-Ras, which leads to Cetuximab resistance, affects Db$\alpha$EGFR-scTRAIL sensitivity, we generated stable Caco-2tet cells inducibly expressing oncogenic K-RasG12V. In the presence of doxycycline, these cells showed increased resistance to Db$\alpha$EGFR-scTRAIL, associated with the elevated expression of the anti-apoptotic proteins cIAP2, Bcl-xL and FlipS. Co-treatment of cells with the Smac mimetic SM83 restored the Db$\alpha$EGFR-scTRAIL-induced apoptotic response. Importantly, this synergy between Db$\alpha$EGFR-scTRAIL and SM83 also translated to 3D cultures of oncogenic K-Ras expressing HCT-116 and LoVo colorectal cancer cells. Our findings thus support the notion that Db$\alpha$EGFR-scTRAIL therapy in combination with apoptosis-sensitizing agents may be promising for the treatment of EGFR-positive colorectal cancers, independently of their KRAS status.}, added-at = {2023-06-29T13:07:55.000+0200}, author = {M{\"{o}}ller, Yvonne and Siegemund, Martin and Beyes, Sven and Herr, Ricarda and Lecis, Daniele and Delia, Domenico and Kontermann, Roland and Brummer, Tilman and Pfizenmaier, Klaus and Olayioye, Monilola A.}, biburl = {https://puma.ub.uni-stuttgart.de/bibtex/24c29ff2df698ac2ee6e5ac658b16a8fb/fabian}, doi = {10.1371/journal.pone.0107165}, editor = {Souglakos, John}, interhash = {3f89113ac48f88a51fecf5b60ce09091}, intrahash = {4c29ff2df698ac2ee6e5ac658b16a8fb}, isbn = {1932-6203 (Electronic)$\backslash$r1932-6203 (Linking)}, issn = {19326203}, journal = {PLoS ONE}, keywords = {2014 izi kontermann olayioye pfizenmaier}, month = sep, number = 9, pages = {e107165}, pmid = {25198428}, timestamp = {2023-06-29T13:07:55.000+0200}, title = {{EGFR-targeted TRAIL and a Smac mimetic synergize to overcome apoptosis resistance in KRAS mutant colorectal cancer cells}}, url = {http://www.ncbi.nlm.nih.gov/pubmed/25198428}, volume = 9, year = 2014 }