Marcus C M Stroet1,2, Bianca M Dijkstra3, Sebastiaan E Dulfer3, Schelto Kruijff4, Wilfred F A den Dunnen5, Frank A E Kruyt6, Rob J M Groen3, Yann Seimbille7, Kranthi M Panth7,8, Laura Mezzanotte7,8, Clemens W G M Lowik7,9, Marion de Jong7. 1. Department of Radiology and Nuclear Medicine/Molecular Genetics, Erasmus Medical Centre, 's-Gravendijkwal 230, 3015 CE, Rotterdam, The Netherlands. m.stroet@erasmusmc.nl. 2. Department of Molecular Genetics, Erasmus MC, Rotterdam, The Netherlands. m.stroet@erasmusmc.nl. 3. Department of Neurosurgery, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands. 4. Department of Surgery, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands. 5. Department of Pathology and Medical Biology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands. 6. Department of Medical Oncology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands. 7. Department of Radiology and Nuclear Medicine/Molecular Genetics, Erasmus Medical Centre, 's-Gravendijkwal 230, 3015 CE, Rotterdam, The Netherlands. 8. Department of Molecular Genetics, Erasmus MC, Rotterdam, The Netherlands. 9. CHUV Department of Oncology, University of Lausanne, Lausanne, Switzerland.
Abstract
BACKGROUND: There is a growing body of nuclear contrast agents that are repurposed for fluorescence-guided surgery. New contrast agents are obtained by substituting the radioactive tag with, or adding a fluorescent cyanine to the molecular structure of antibodies or peptides. This enables intra-operative fluorescent detection of cancerous tissue, leading to more complete tumor resection. However, these fluorescent cyanines can have a remarkable influence on pharmacokinetics and tumor uptake, especially when labeled to smaller targeting vectors such as peptides. Here we demonstrate the effect of cyanine-mediated dead cell-binding of Ac-Lys0(IRDye800CW)-Tyr3-octreotate (800CW-TATE) and how this can be used as an advantage for fluorescence-guided surgery. RESULTS: Binding of 800CW-TATE could be blocked with DOTA0-Tyr3-octreotate (DOTA-TATE) on cultured SSTR2-positive U2OS cells and was absent in SSTR2 negative U2OS cells. However, strong binding was observed to dead cells, which could not be blocked with DOTA-TATE and was also present in dead SSTR2 negative cells. No SSTR2-mediated binding was observed in frozen tumor sections, possibly due to disruption of the cells in the process of sectioning the tissue before exposure to the contrast agent. DOTA-TATE blocking resulted in an incomplete reduction of 61.5 ± 5.8% fluorescence uptake by NCI-H69-tumors in mice. Near-infrared imaging and dead cell staining on paraffin sections from resected tumors revealed that fluorescence uptake persisted in necrotic regions upon blocking with DOTA-TATE. CONCLUSION: This study shows that labeling peptides with cyanines can result in dead cell binding. This does not hamper the ultimate purpose of fluorescence-guided surgery, as necrotic tissue appears in most solid tumors. Hence, the necrosis binding can increase the overall tumor uptake. Moreover, necrotic tissue should be removed as much as possible: it cannot be salvaged, causes inflammation, and is tumorigenic. However, when performing binding experiments to cells with disrupted membrane integrity, which is routinely done with nuclear probes, this dead cell-binding can resemble non-specific binding. This study will benefit the development of fluorescent contrast agents.
BACKGROUND: There is a growing body of nuclear contrast agents that are repurposed for fluorescence-guided surgery. New contrast agents are obtained by substituting the radioactive tag with, or adding a fluorescent cyanine to the molecular structure of antibodies or peptides. This enables intra-operative fluorescent detection of cancerous tissue, leading to more complete tumor resection. However, these fluorescent cyanines can have a remarkable influence on pharmacokinetics and tumor uptake, especially when labeled to smaller targeting vectors such as peptides. Here we demonstrate the effect of cyanine-mediated dead cell-binding of Ac-Lys0(IRDye800CW)-Tyr3-octreotate (800CW-TATE) and how this can be used as an advantage for fluorescence-guided surgery. RESULTS: Binding of 800CW-TATE could be blocked with DOTA0-Tyr3-octreotate (DOTA-TATE) on cultured SSTR2-positive U2OS cells and was absent in SSTR2 negative U2OS cells. However, strong binding was observed to dead cells, which could not be blocked with DOTA-TATE and was also present in dead SSTR2 negative cells. No SSTR2-mediated binding was observed in frozen tumor sections, possibly due to disruption of the cells in the process of sectioning the tissue before exposure to the contrast agent. DOTA-TATE blocking resulted in an incomplete reduction of 61.5 ± 5.8% fluorescence uptake by NCI-H69-tumors in mice. Near-infrared imaging and dead cell staining on paraffin sections from resected tumors revealed that fluorescence uptake persisted in necrotic regions upon blocking with DOTA-TATE. CONCLUSION: This study shows that labeling peptides with cyanines can result in dead cell binding. This does not hamper the ultimate purpose of fluorescence-guided surgery, as necrotic tissue appears in most solid tumors. Hence, the necrosis binding can increase the overall tumor uptake. Moreover, necrotic tissue should be removed as much as possible: it cannot be salvaged, causes inflammation, and is tumorigenic. However, when performing binding experiments to cells with disrupted membrane integrity, which is routinely done with nuclear probes, this dead cell-binding can resemble non-specific binding. This study will benefit the development of fluorescent contrast agents.
Authors: Pieter J Steinkamp; Bobby K Pranger; Mei-Fang Li; Matthijs D Linssen; Floris J Voskuil; Lukas B Been; Barbara L van Leeuwen; Albert J H Suurmeijer; Wouter B Nagengast; Schelto Kruijff; Robert J van Ginkel; Gooitzen M van Dam Journal: J Nucl Med Date: 2020-07-17 Impact factor: 10.057
Authors: Eva J Ter Weele; Anton G T Terwisscha van Scheltinga; Matthijs D Linssen; Wouter B Nagengast; Ingo Lindner; Annelies Jorritsma-Smit; Elisabeth G E de Vries; Jos G W Kosterink; Marjolijn N Lub-de Hooge Journal: Eur J Pharm Biopharm Date: 2016-05-12 Impact factor: 5.571
Authors: Willemieke S Tummers; Sarah E Miller; Nutte T Teraphongphom; Adam Gomez; Idan Steinberg; David M Huland; Steve Hong; Sri-Rajasekhar Kothapalli; Alifia Hasan; Robert Ertsey; Bert A Bonsing; Alexander L Vahrmeijer; Rutger-Jan Swijnenburg; Teri A Longacre; George A Fisher; Sanjiv S Gambhir; George A Poultsides; Eben L Rosenthal Journal: Ann Surg Oncol Date: 2018-04-17 Impact factor: 5.344
Authors: Bangwen Xie; Marieke A Stammes; Pieter B A A van Driel; Luis J Cruz; Vicky T Knol-Blankevoort; Martijn A M Löwik; Laura Mezzanotte; Ivo Que; Alan Chan; Jeroen P H M van den Wijngaard; Maria Siebes; Sven Gottschalk; Daniel Razansky; Vasilis Ntziachristos; Stijn Keereweer; Richard W Horobin; Mathias Hoehn; Eric L Kaijzel; Ermond R van Beek; Thomas J A Snoeks; Clemens W G M Löwik Journal: Oncotarget Date: 2015-11-17
Authors: Marieke A Stammes; Vicky T Knol-Blankevoort; Luis J Cruz; Hans R I J Feitsma; Laura Mezzanotte; Robert A Cordfunke; Riccardo Sinisi; Elena A Dubikovskaya; Azusa Maeda; Ralph S DaCosta; Katja Bierau; Alan Chan; Eric L Kaijzel; Thomas J A Snoeks; Ermond R van Beek; Clemens W G M Löwik Journal: Mol Imaging Biol Date: 2016-12 Impact factor: 3.488
Authors: Marieke A Stammes; Azusa Maeda; Jiachuan Bu; Deborah A Scollard; Iris Kulbatski; Philip J Medeiros; Riccardo Sinisi; Elena A Dubikovskaya; Thomas J A Snoeks; Ermond R van Beek; Alan B Chan; Clemens W G M Löwik; Ralph S DaCosta Journal: Front Oncol Date: 2016-10-21 Impact factor: 6.244
Authors: Fijs W B van Leeuwen; Bart Cornelissen; Federico Caobelli; Laura Evangelista; Latifa Rbah-Vidal; Silvana Del Vecchio; Catarina Xavier; Jacques Barbet; Marion de Jong Journal: EJNMMI Radiopharm Chem Date: 2017-12-15
Authors: Marcus C M Stroet; Erik de Blois; Debra C Stuurman; Corrina M A de Ridder; Joost Haeck; Yann Seimbille; Laura Mezzanotte; Marion de Jong; Clemens W G M Löwik; Kranthi M Panth Journal: Mol Imaging Biol Date: 2020-10 Impact factor: 3.488
Authors: Marcus C M Stroet; Erik de Blois; Marion de Jong; Yann Seimbille; Laura Mezzanotte; Clemens W G M Löwik; Kranthi M Panth Journal: Cancers (Basel) Date: 2022-02-09 Impact factor: 6.639
Authors: Syed Muhammad Usama; Sierra C Marker; Servando Hernandez Vargas; Solmaz AghaAmiri; Sukhen C Ghosh; Naruhiko Ikoma; Hop S Tran Cao; Martin J Schnermann; Ali Azhdarinia Journal: Cancers (Basel) Date: 2022-03-23 Impact factor: 6.639