title: Hypoxia-Targeting Fluorescent Nanobodies for Optical Molecular Imaging of Pre-Invasive Breast Cancer.
abstract: The aim of this work was to develop a CAIX-specific nanobody conjugated to IRDye800CW for molecular imaging of pre-invasive breast cancer.
  
  
  
  CAIX-specific nanobodies were selected using a modified phage display technology, conjugated site-specifically to IRDye800CW and evaluated in a xenograft breast cancer mouse model using ductal carcinoma in situ cells (DCIS).
  
  
  
  Specific anti-CAIX nanobodies were obtained. Administration of a CAIX-specific nanobody into mice with DCIS xenografts overexpressing CAIX showed after 2 h a mean tumor-to-normal tissue ratio (TNR) of 4.3 ± 0.6, compared to a TNR of 1.4 ± 0.2 in mice injected with the negative control nanobody R2-IR. In DCIS mice, a TNR of 1.8 ± 0.1 was obtained. Biodistribution studies demonstrated an uptake of 14.0 ± 1.1 %I.D./g in DCIS + CAIX tumors, 4.6 ± 0.8 %I.D./g in DCIS tumors, while 2.0 ± 0.2 %I.D./g was obtained with R2-IR.
  
  
  
  These results demonstrate the successful generation of a CAIX-specific nanobody-IRDye800CW conjugate that can be used for rapid imaging of (pre-)invasive breast cancer.
authors.affiliation: Division of Cell Biology, Department of Biology, Science Faculty, Utrecht University, Padualaan 8, 3584 CH, Utrecht, The Netherlands.|Department of Pathology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: Aram S A
authors.initials: AS
authors.lastname: van Brussel
authors.affiliation: Department of Radiology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: Arthur
authors.initials: A
authors.lastname: Adams
authors.affiliation: Division of Cell Biology, Department of Biology, Science Faculty, Utrecht University, Padualaan 8, 3584 CH, Utrecht, The Netherlands.|Department of Pathology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Oliveira
authors.affiliation: Division of Cell Biology, Department of Biology, Science Faculty, Utrecht University, Padualaan 8, 3584 CH, Utrecht, The Netherlands.
authors.forename: Bram
authors.initials: B
authors.lastname: Dorresteijn
authors.affiliation: QVQ BV, Utrecht, The Netherlands.
authors.forename: Mohamed
authors.initials: M
authors.lastname: El Khattabi
authors.affiliation: Department of Pathology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: Jeroen F
authors.initials: JF
authors.lastname: Vermeulen
authors.affiliation: Division of Internal Medicine and Dermatology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: Elsken
authors.initials: E
authors.lastname: van der Wall
authors.affiliation: Department of Radiology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: Willem P Th M
authors.initials: WP
authors.lastname: Mali
authors.affiliation: Department of Pathology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: Patrick W B
authors.initials: PW
authors.lastname: Derksen
authors.affiliation: Department of Pathology, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: Paul J
authors.initials: PJ
authors.lastname: van Diest
authors.affiliation: Division of Cell Biology, Department of Biology, Science Faculty, Utrecht University, Padualaan 8, 3584 CH, Utrecht, The Netherlands. p.vanbergen@uu.nl.
authors.forename: Paul M P
authors.initials: PM
authors.lastname: van Bergen En Henegouwen
mesh_terms: D000818:Animals; D000071231:Carbonic Anhydrase IX / Q000378:metabolism; D002285:Carcinoma, Intraductal, Noninfiltrating / Q000473:pathology; D015687:Cell Hypoxia; D061505:Cell Surface Display Techniques; D004247:DNA / Q000378:metabolism; D005260:Female; D006367:HeLa Cells; D006801:Humans; D000860:Hypoxia / Q000473:pathology*; D007114:Immunization; D007150:Immunohistochemistry; D015674:Mammary Neoplasms, Animal / Q000473:pathology*; D051379:Mice; D057054:Molecular Imaging / Q000379:methods*; D009361:Neoplasm Invasiveness; D061848:Optical Imaging / Q000379:methods*; D061905:Single-Domain Antibodies / Q000378:metabolism*; D014018:Tissue Distribution; D023041:Xenograft Model Antitumor Assays
chemical_list: D061905:Single-Domain Antibodies; D004247:DNA; D000071231:Carbonic Anhydrase IX
keywords: Breast cancer; Carbonic anhydrase IX; Molecular fluorescence pathology; Nanobody; Optical imaging; VHH