TY - JOUR
T1 - Molecular imaging with a fluorescent antibody targeting carbonic anhydrase IX can successfully detect hypoxic ductal carcinoma in situ of the breast
AU - Van Brussel, Aram S.A.
AU - Adams, Arthur
AU - Vermeulen, Jeroen F.
AU - Oliveira, Sabrina
AU - Van Der Wall, Elsken
AU - Mali, Willem P.Th M.
AU - Van Diest, Paul J.
AU - Van Bergen En Henegouwen, Paul M.P.
N1 - Funding Information:
Acknowledgments We would like to thank A. Martens for the pLV-CMV-LUC2-IRES-GFP vector and P.C. Pearlman for writing the algorithm in Matlab. We are indebted to P.W.B. Derksen and S.G. Elias for their suggestions concerning the mouse model. Also, we would like to thank the animal facility of the University Utrecht and the biobank of the University Medical Center Utrecht for their support. This research was supported by the Center for Translational Molecular Medicine—Mammary Carcinoma Molecular Imaging for Diagnosis and Therapeutics (CTMM—MAMMOTH, Project 203)— and by an unrestricted educational grant from Aegon to P.J.vD.
Copyright:
Copyright 2013 Elsevier B.V., All rights reserved.
PY - 2013/7
Y1 - 2013/7
N2 - Ductal carcinoma in situ (DCIS) of the breast is difficult to remove completely during surgery as it is not palpable and can therefore require re-excision. Real-time visualization of DCIS using near-infrared fluorescent probes could help the surgeon during surgery as well as the pathologist post-operatively to distinguish the tumor from healthy tissue. As hypoxia-induced necrosis is a common phenomenon in DCIS, we investigated the molecular imaging of DCIS using a fluorescent antibody targeting a hypoxia marker, carbonic anhydrase IX (CAIX), in a preclinical mouse model. A monoclonal antibody against human CAIX was fluorescently labeled with the near-infrared dye IRDye800CW and characterized in vitro. An in vivo study was performed in SCID/Beige mice that were orthotopically transplanted with human breast cancer cells mimicking human DCIS (MCF10DCIS) and MCF10DCIS stably expressing CAIX. A clinically approved fluorescence imaging system was used to monitor probe uptake and to determine tumor-to-normal tissue ratios (TNR). Mean in vivo TNR of CAIX-transduced (CAIX+) tumors was 7.5 ± 0.5. Mean in vivo TNR of DCIS tumors with hypoxic areas reached a plateau level at 48 h after injection of 2.1 ± 0.1 (mean ± SEM) compared to 1.7 ± 0.1 in DCIS without hypoxic areas. Mean intra-operative TNR of DCIS tumors with necrotic regions was higher than that of DCIS tumors without necrotic regions 96 h after injection - 2.9 ± 0.1 and 1.5 ± 0.1, respectively - while the TNR of CAIX+ tumors was 11.2 ± 1.0. Specific tumor uptake of MabCAIX-IRDye800CW was confirmed by a biodistribution assay, and immunofluorescence imaging on tumor sections showed specific uptake in hypoxic tumor regions, with higher contrast than conventional chromagen-based immunohistochemistry. Molecular fluorescence imaging with MabCAIX-IRDye800CW can be successfully used to detect hypoxic DCIS before and during surgery to facilitate radical resection. Furthermore, it allows for sensitive CAIX-specific immunofluorescence microscopy of tumor sections, thereby introducing the concept of molecular fluorescence pathology.
AB - Ductal carcinoma in situ (DCIS) of the breast is difficult to remove completely during surgery as it is not palpable and can therefore require re-excision. Real-time visualization of DCIS using near-infrared fluorescent probes could help the surgeon during surgery as well as the pathologist post-operatively to distinguish the tumor from healthy tissue. As hypoxia-induced necrosis is a common phenomenon in DCIS, we investigated the molecular imaging of DCIS using a fluorescent antibody targeting a hypoxia marker, carbonic anhydrase IX (CAIX), in a preclinical mouse model. A monoclonal antibody against human CAIX was fluorescently labeled with the near-infrared dye IRDye800CW and characterized in vitro. An in vivo study was performed in SCID/Beige mice that were orthotopically transplanted with human breast cancer cells mimicking human DCIS (MCF10DCIS) and MCF10DCIS stably expressing CAIX. A clinically approved fluorescence imaging system was used to monitor probe uptake and to determine tumor-to-normal tissue ratios (TNR). Mean in vivo TNR of CAIX-transduced (CAIX+) tumors was 7.5 ± 0.5. Mean in vivo TNR of DCIS tumors with hypoxic areas reached a plateau level at 48 h after injection of 2.1 ± 0.1 (mean ± SEM) compared to 1.7 ± 0.1 in DCIS without hypoxic areas. Mean intra-operative TNR of DCIS tumors with necrotic regions was higher than that of DCIS tumors without necrotic regions 96 h after injection - 2.9 ± 0.1 and 1.5 ± 0.1, respectively - while the TNR of CAIX+ tumors was 11.2 ± 1.0. Specific tumor uptake of MabCAIX-IRDye800CW was confirmed by a biodistribution assay, and immunofluorescence imaging on tumor sections showed specific uptake in hypoxic tumor regions, with higher contrast than conventional chromagen-based immunohistochemistry. Molecular fluorescence imaging with MabCAIX-IRDye800CW can be successfully used to detect hypoxic DCIS before and during surgery to facilitate radical resection. Furthermore, it allows for sensitive CAIX-specific immunofluorescence microscopy of tumor sections, thereby introducing the concept of molecular fluorescence pathology.
KW - Carbonic anhydrase IX
KW - DCIS
KW - Hypoxia
KW - Molecular imaging
KW - Monoclonal antibody
KW - Near-infrared
UR - http://www.scopus.com/inward/record.url?scp=84881481421&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84881481421&partnerID=8YFLogxK
U2 - 10.1007/s10549-013-2635-6
DO - 10.1007/s10549-013-2635-6
M3 - Article
C2 - 23860929
AN - SCOPUS:84881481421
SN - 0167-6806
VL - 140
SP - 263
EP - 272
JO - Breast Cancer Research and Treatment
JF - Breast Cancer Research and Treatment
IS - 2
ER -