TY - JOUR
T1 - AD-O53.2 - A novel recombinant fusion protein combining the activities of TRAIL/Apo2L and Smac/Diablo, overcomes resistance of human cancer cells to TRAIL/Apo2L
AU - Pieczykolan, Jerzy S.
AU - Kubiński, Konrad
AU - Masłyk, Maciej
AU - Pawlak, Sebastian D.
AU - Pieczykolan, Anna
AU - Rózga, Piotr K.
AU - Szymanik, Michał
AU - Gałązka, Marlena
AU - Teska-Kamińska, Małgorzata
AU - Żerek, Bartłomiej
AU - Bukato, Katarzyna
AU - Poleszak, Katarzyna
AU - Jaworski, Albert
AU - Strożek, Wojciech
AU - ͆wider, Robert
AU - Zieliński, Rafał
N1 - Funding Information:
The work was financially supported: “3CLA—biotechnological aimed anticancer drug”. Key project—no. POIG.01.94.00-00-002/08, fulfilled as a part of Programme “Innovative Economy”, Priority 1. Research and development of modern technologies, Action 1.4. Financed by European Funds for Regional Development—85 %, government budget—15 %, years: 2008–2014.
Publisher Copyright:
© 2014 Springer Science+Business Media New York.
PY - 2014/12/1
Y1 - 2014/12/1
N2 - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and its receptors became promising molecules for selective targeting of tumor cells without affecting normal tissue. Unfortunately, cancer cells have developed a number of mechanisms that confer resistance to TRAIL\Apo2L-induced apoptosis, which substantiates the need for development of alternative therapeutic strategies. Here we present a recombinant variant of TRAIL\Apo2L peptide, named AD-O53.2, fused to the peptide-derived from Smac/Diablo protein - the natural inhibitor of the apoptotic X-linked IAP (XIAP) protein considered as a pro-apoptotic agent. The proposed mechanism of action for this construct involves specific targeting of the tumor by TRAIL\Apo2L followed by activation and internalization of pro-apoptotic peptide into the cancer cells. While in the cytoplasm, the Smac\Diablo peptide inhibits activity of X-linked IAP (XIAP) proteins and promotes caspase-mediated apoptosis. AD-O53.2 construct was expressed in E.coli and purified by Ion Exchange Chromatography (IEC). Derived protein was initially characterized by circular dichroism spectroscopy (CD), HPLC-SEC chromatography, surface plasmon resonance, protease activation and cell proliferation assays. Our Smac/Diablo-TRAIL fusion variant was tested against a panel of cancer cells (including lung, colorectal, pancreatic, liver, kidney and uterine) and showed a potent cytotoxic effect with the IC 50 values in femtomolar range for the most sensitive cell lines, while it remained ineffective against non-transformed HUVEC cells as well as isolated normal human and rat hepatocytes. Importantly, the construct was well tolerated by animals and significantly reduced the rate of the tumor growth in colon and lung adenocarcinoma animal models.
AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and its receptors became promising molecules for selective targeting of tumor cells without affecting normal tissue. Unfortunately, cancer cells have developed a number of mechanisms that confer resistance to TRAIL\Apo2L-induced apoptosis, which substantiates the need for development of alternative therapeutic strategies. Here we present a recombinant variant of TRAIL\Apo2L peptide, named AD-O53.2, fused to the peptide-derived from Smac/Diablo protein - the natural inhibitor of the apoptotic X-linked IAP (XIAP) protein considered as a pro-apoptotic agent. The proposed mechanism of action for this construct involves specific targeting of the tumor by TRAIL\Apo2L followed by activation and internalization of pro-apoptotic peptide into the cancer cells. While in the cytoplasm, the Smac\Diablo peptide inhibits activity of X-linked IAP (XIAP) proteins and promotes caspase-mediated apoptosis. AD-O53.2 construct was expressed in E.coli and purified by Ion Exchange Chromatography (IEC). Derived protein was initially characterized by circular dichroism spectroscopy (CD), HPLC-SEC chromatography, surface plasmon resonance, protease activation and cell proliferation assays. Our Smac/Diablo-TRAIL fusion variant was tested against a panel of cancer cells (including lung, colorectal, pancreatic, liver, kidney and uterine) and showed a potent cytotoxic effect with the IC 50 values in femtomolar range for the most sensitive cell lines, while it remained ineffective against non-transformed HUVEC cells as well as isolated normal human and rat hepatocytes. Importantly, the construct was well tolerated by animals and significantly reduced the rate of the tumor growth in colon and lung adenocarcinoma animal models.
KW - Apoptosis
KW - Cell resistance
KW - Cytotoxicity
KW - Fusion protein
KW - TRAIL/Apo2L
UR - http://www.scopus.com/inward/record.url?scp=84927563731&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84927563731&partnerID=8YFLogxK
U2 - 10.1007/s10637-014-0153-y
DO - 10.1007/s10637-014-0153-y
M3 - Article
C2 - 25182378
AN - SCOPUS:84927563731
SN - 0167-6997
VL - 32
SP - 1155
EP - 1166
JO - Investigational New Drugs
JF - Investigational New Drugs
IS - 6
ER -