Kai-Wen Ho1,2,3,4, I-J U Chen2,3,4, Yi-An Cheng2,3,4, Tzu-Yi Liao1,2,4, En-Shuo Liu2,3,4, Huei-Jen Chen1,2,4, Yun-Chi Lu2,3,4, Yu-Cheng Su4,5, Steve R Roffler1,4,6, Bo-Cheng Huang4,7, Hui-Ju Liu1,2,4, Ming-Yii Huang1,4,8,9, Chiao-Yun Chen10,11,12,13, Tian-Lu Cheng14,15,16,17,18. 1. Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan. 2. Drug Development and Value Creation Research Center, Kaohsiung Medical University, Kaohsiung, Taiwan. 3. Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University, No. 100 Shih-Chuan 1st Road, Kaohsiung, 80708, Taiwan. 4. Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University, Kaohsiung, Taiwan. 5. Institute of Molecular Medicine and Bioengineering, Department of Biological Science and Technology, National Chiao Tung University, Hsin-Chu, Taiwan. 6. Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan. 7. Institute of Biomedical Sciences, National Sun Yat-Sen University, Kaohsiung, Taiwan. 8. Department of Radiation Oncology, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan. 9. Department of Radiation Oncology, Faculty of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan. 10. Drug Development and Value Creation Research Center, Kaohsiung Medical University, Kaohsiung, Taiwan. ccy77015@gmail.com. 11. Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University, Kaohsiung, Taiwan. ccy77015@gmail.com. 12. Department of Radiation Oncology, Faculty of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan. ccy77015@gmail.com. 13. Department of Medical Imaging, Kaohsiung Medical University Hospital, Sanmin Dist, No.100, Tzyou 1st Rd, Kaohsiung, Taiwan. ccy77015@gmail.com. 14. Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan. tlcheng@kmu.edu.tw. 15. Drug Development and Value Creation Research Center, Kaohsiung Medical University, Kaohsiung, Taiwan. tlcheng@kmu.edu.tw. 16. Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University, No. 100 Shih-Chuan 1st Road, Kaohsiung, 80708, Taiwan. tlcheng@kmu.edu.tw. 17. Department of Medical Research, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan. tlcheng@kmu.edu.tw. 18. Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University, Kaohsiung, Taiwan. tlcheng@kmu.edu.tw.
Abstract
BACKGROUND: Tumor-targeted nanoparticles hold great promise as new tools for therapy of liquid cancers. Furthermore, the therapeutic efficacy of nanoparticles can be improved by enhancing the cancer cellular internalization. METHODS: In this study, we developed a humanized bispecific antibody (BsAbs: CD20 Ab-mPEG scFv) which retains the clinical anti-CD20 whole antibody (Ofatumumab) and is fused with an anti-mPEG single chain antibody (scFv) that can target the systemic liquid tumor cells. This combination achieves the therapeutic function and simultaneously "grabs" Lipo-Dox® (PEGylated liposomal doxorubicin, PLD) to enhance the cellular internalization and anticancer activity of PLD. RESULTS: We successfully constructed the CD20 Ab-mPEG scFv and proved that CD20 Ab-mPEG scFv can target CD20-expressing Raji cells and simultaneously grab PEGylated liposomal DiD increasing the internalization ability up to 60% in 24 h. We further showed that the combination of CD20 Ab-mPEG scFv and PLD successfully led to a ninefold increase in tumor cytotoxicity (LC50: 0.38 nM) compared to the CD20 Ab-DNS scFv and PLD (lC50: 3.45 nM) in vitro. Importantly, a combination of CD20 Ab-mPEG scFv and PLD had greater anti-liquid tumor efficacy (P = 0.0005) in Raji-bearing mice than CD20 Ab-DNS scFv and PLD. CONCLUSION: Our results indicate that this "double-attack" strategy using CD20 Ab-mPEG scFv and PLD can retain the tumor targeting (first attack) and confer PLD tumor-selectivity (second attack) to enhance PLD internalization and improve therapeutic efficacy in liquid tumors.
BACKGROUND:Tumor-targeted nanoparticles hold great promise as new tools for therapy of liquid cancers. Furthermore, the therapeutic efficacy of nanoparticles can be improved by enhancing the cancer cellular internalization. METHODS: In this study, we developed a humanized bispecific antibody (BsAbs: CD20 Ab-mPEG scFv) which retains the clinical anti-CD20 whole antibody (Ofatumumab) and is fused with an anti-mPEG single chain antibody (scFv) that can target the systemic liquid tumor cells. This combination achieves the therapeutic function and simultaneously "grabs" Lipo-Dox® (PEGylated liposomal doxorubicin, PLD) to enhance the cellular internalization and anticancer activity of PLD. RESULTS: We successfully constructed the CD20 Ab-mPEG scFv and proved that CD20 Ab-mPEG scFv can target CD20-expressing Raji cells and simultaneously grab PEGylated liposomal DiD increasing the internalization ability up to 60% in 24 h. We further showed that the combination of CD20 Ab-mPEG scFv and PLD successfully led to a ninefold increase in tumor cytotoxicity (LC50: 0.38 nM) compared to the CD20Ab-DNS scFv and PLD (lC50: 3.45 nM) in vitro. Importantly, a combination of CD20 Ab-mPEG scFv and PLD had greater anti-liquid tumor efficacy (P = 0.0005) in Raji-bearing mice than CD20Ab-DNS scFv and PLD. CONCLUSION: Our results indicate that this "double-attack" strategy using CD20 Ab-mPEG scFv and PLD can retain the tumor targeting (first attack) and confer PLD tumor-selectivity (second attack) to enhance PLD internalization and improve therapeutic efficacy in liquid tumors.
Authors: Carla Casulo; Michelle Byrtek; Keith L Dawson; Xiaolei Zhou; Charles M Farber; Christopher R Flowers; John D Hainsworth; Matthew J Maurer; James R Cerhan; Brian K Link; Andrew D Zelenetz; Jonathan W Friedberg Journal: J Clin Oncol Date: 2015-06-29 Impact factor: 44.544