Thinzar M Lwin1, Takashi Murakami1,2,3, Kentaro Miyake1,2,3, Paul J Yazaki4, John E Shivley4, Robert M Hoffman1,2, Michael Bouvet5,6,7. 1. Department of Surgery, University of California San Diego, San Diego, CA, USA. 2. AntiCancer, Inc., San Diego, CA, USA. 3. Department of Gastroenterological Surgery, Graduate School of Medicine, Yokohama City University, Yokohama, Japan. 4. City of Hope National Medical Center, Duarte, CA, USA. 5. Department of Surgery, University of California San Diego, San Diego, CA, USA. mbouvet@ucsd.edu. 6. VA San Diego Healthcare System, San Diego, CA, USA. mbouvet@ucsd.edu. 7. Moores Cancer Center, University of California San Diego, San Diego, CA, USA. mbouvet@ucsd.edu.
Abstract
BACKGROUND/ PURPOSE: Development of a humanized fluorophore-conjugated antibody that can improve contrast for fluorescence-guided oncologic surgeries. METHODS: BxPC-3-GFP pancreatic cancer cells were injected into flanks of nude mice. Fragments of subcutaneous tumors were grafted onto the pancreatic tail of recipient mice to create orthotopic xenograft models of pancreatic cancer. After tumors developed for 4 weeks, a humanized anti-carcinoembryonic antigen antibody conjugated to an 800 nm near-infrared fluorescent dye (hM5A-IR800) was injected intravenously. Mice were imaged at 6, 12, 24, 48, and 72 h after injection. RESULTS: Fluorescence imaging showed that hM5A-IR800 specifically localized to BxPC-3 human pancreatic cancer cells. The fluorescent probe localized to cell surfaces in vitro and specifically co-localized with green fluorescent protein-labeled tumors in an orthotopic pancreatic xenograft model in vivo. Serial imaging at specific time points showed peak signal intensity of the orthotopic pancreatic tumor at 48 h; this time point corresponded with a maximal tumor-to-background ratio (TBR) of 16.6 at 48 h. DISCUSSION: hM5A-IR800 was successfully able to specifically label orthotopic pancreatic tumors in situ. The longer wavelength allowed deeper tissue penetration, particularly in tumor areas covered by normal pancreatic parenchyma. The probe had expected kinetics for an antibody-fluorophore conjugate, with the peak signal intensity reached at 48 h. A clear tumor signal was observed with a TBR > 5 at all time points, with high contrast (TBR of 16.6) at 48 h. CONCLUSION: hM5A-IR800 demonstrated excellent tumor localization and a very bright signal. It is a promising agent for future clinical fluorescence-guided surgery applications.
BACKGROUND/ PURPOSE: Development of a humanized fluorophore-conjugated antibody that can improve contrast for fluorescence-guided oncologic surgeries. METHODS:BxPC-3-GFP pancreatic cancer cells were injected into flanks of nude mice. Fragments of subcutaneous tumors were grafted onto the pancreatic tail of recipient mice to create orthotopic xenograft models of pancreatic cancer. After tumors developed for 4 weeks, a humanized anti-carcinoembryonic antigen antibody conjugated to an 800 nm near-infrared fluorescent dye (hM5A-IR800) was injected intravenously. Mice were imaged at 6, 12, 24, 48, and 72 h after injection. RESULTS: Fluorescence imaging showed that hM5A-IR800 specifically localized to BxPC-3humanpancreatic cancer cells. The fluorescent probe localized to cell surfaces in vitro and specifically co-localized with green fluorescent protein-labeled tumors in an orthotopic pancreatic xenograft model in vivo. Serial imaging at specific time points showed peak signal intensity of the orthotopic pancreatic tumor at 48 h; this time point corresponded with a maximal tumor-to-background ratio (TBR) of 16.6 at 48 h. DISCUSSION: hM5A-IR800 was successfully able to specifically label orthotopic pancreatic tumors in situ. The longer wavelength allowed deeper tissue penetration, particularly in tumor areas covered by normal pancreatic parenchyma. The probe had expected kinetics for an antibody-fluorophore conjugate, with the peak signal intensity reached at 48 h. A clear tumor signal was observed with a TBR > 5 at all time points, with high contrast (TBR of 16.6) at 48 h. CONCLUSION:hM5A-IR800 demonstrated excellent tumor localization and a very bright signal. It is a promising agent for future clinical fluorescence-guided surgery applications.
Authors: Ali A Maawy; Yukihiko Hiroshima; Sharmeela Kaushal; George A Luiken; Robert M Hoffman; Michael Bouvet Journal: J Biomed Opt Date: 2013-12 Impact factor: 3.170
Authors: Oliver Strobel; Thomas Hank; Ulf Hinz; Frank Bergmann; Lutz Schneider; Christoph Springfeld; Dirk Jäger; Peter Schirmacher; Thilo Hackert; Markus W Büchler Journal: Ann Surg Date: 2017-03 Impact factor: 12.969
Authors: Sharmeela Kaushal; Michele K McElroy; George A Luiken; Mark A Talamini; A R Moossa; Robert M Hoffman; Michael Bouvet Journal: J Gastrointest Surg Date: 2008-07-30 Impact factor: 3.452
Authors: Cristina A Metildi; Chih-Min Tang; Sharmeela Kaushal; Stephanie Y Leonard; Paolo Magistri; Hop S Tran Cao; Robert M Hoffman; Michael Bouvet; Jason K Sicklick Journal: Ann Surg Oncol Date: 2013-08-14 Impact factor: 5.344
Authors: Leonora S F Boogerd; Henricus J M Handgraaf; Hwai-Ding Lam; Volkert A L Huurman; Arantza Farina-Sarasqueta; John V Frangioni; Cornelis J H van de Velde; Andries E Braat; Alexander L Vahrmeijer Journal: Surg Endosc Date: 2016-06-29 Impact factor: 4.584
Authors: Ekaterina Mikhaylova; Jamison Brooks; Darren Zuro; Farouk Nouizi; Maciej Kujawski; Srideshikan Sargur Madabushi; Jinyi Qi; Mengxi Zhang; Junie Chea; Erasmus K Poku; Nicole Bowles; Jeffrey Y C Wong; John E Shively; Paul J Yazaki; Gultekin Gulsen; Simon R Cherry; Susanta Hui Journal: IEEE Access Date: 2019-09-30 Impact factor: 3.367
Authors: Thinzar M Lwin; Sophie Hernot; Hannah Hollandsworth; Siamak Amirfakhri; Filemoni Filemoni; Pieterjan Debie; Robert M Hoffman; Michael Bouvet Journal: Surgery Date: 2020-05-04 Impact factor: 3.982
Authors: Thinzar M Lwin; Kentaro Miyake; Takashi Murakami; Jonathan C DeLong; Siamak Amirfakhri; Filemoni Filemoni; Sang Nam Yoon; Paul J Yazaki; John E Shivley; Brian Datnow; Bryan M Clary; Robert M Hoffman; Michael Bouvet Journal: Oncotarget Date: 2018-12-18