title: Clostridium perfringens enterotoxin C-terminal domain labeled to fluorescent dyes for in vivo visualization of micrometastatic chemotherapy-resistant ovarian cancer.
abstract: Identification of micrometastatic disease at the time of surgery remains extremely challenging in ovarian cancer patients. We used fluorescence microscopy, an in vivo imaging system and a fluorescence stereo microscope to evaluate fluorescence distribution in Claudin-3- and -4-overexpressing ovarian tumors, floating tumor clumps isolated from ascites and healthy organs. To do so, mice harboring chemotherapy-naïve and chemotherapy-resistant human ovarian cancer xenografts or patient-derived xenografts (PDXs) were treated with the carboxyl-terminal binding domain of the Clostridium perfringens enterotoxin (c-CPE) conjugated to FITC (FITC-c-CPE) or the near-infrared (NIR) fluorescent tag IRDye CW800 (CW800-c-CPE) either intraperitoneally (IP) or intravenously (IV). We found tumor fluorescence to plateau at 30 min after IP injection of both the FITC-c-CPE and the CW800-c-CPE peptides and to be significantly higher than in healthy organs (p < 0.01). After IV injection of CW800-c-CPE, tumor fluorescence plateaued at 6 hr while the most favorable tumor-to-background fluorescence ratio (TBR) was found at 48 hr in both mouse models. Importantly, fluorescent c-CPE was highly sensitive for the in vivo visualization of peritoneal micrometastatic tumor implants and the identification of ovarian tumor spheroids floating in malignant ascites that were otherwise not detectable by conventional visual observation. The use of the fluorescent c-CPE peptide may represent a novel and effective optical approach at the time of primary debulking surgery for the real-time detection of micrometastatic ovarian disease overexpressing the Claudin-3 and -4 receptors or the identification of residual disease at the time of interval debulking surgery after neoadjuvant chemotherapy treatment.
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT.|Department of Molecular and Translational Medicine, Palazzetto Polifunzionale, Brescia, Italy.
authors.forename: Emiliano
authors.initials: E
authors.lastname: Cocco
authors.affiliation: Department of Radiology, Michigan State University, East Lansing, MI.
authors.forename: Erik M
authors.initials: EM
authors.lastname: Shapiro
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT.
authors.forename: Sara
authors.initials: S
authors.lastname: Gasparrini
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT.|Division of Gynecologic Oncology, University Campus Bio-Medico of Rome, Rome, Italy.
authors.forename: Salvatore
authors.initials: S
authors.lastname: Lopez
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT.
authors.forename: Carlton L
authors.initials: CL
authors.lastname: Schwab
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT.
authors.forename: Stefania
authors.initials: S
authors.lastname: Bellone
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT.
authors.forename: Ileana
authors.initials: I
authors.lastname: Bortolomai
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT.
authors.forename: Natalia J
authors.initials: NJ
authors.lastname: Sumi
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT.
authors.forename: Elena
authors.initials: E
authors.lastname: Bonazzoli
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT.
authors.forename: Roberta
authors.initials: R
authors.lastname: Nicoletti
authors.affiliation: Department of Biomedical Engineering, Yale University, New Haven, CT.
authors.forename: Yang
authors.initials: Y
authors.lastname: Deng
authors.affiliation: Department of Biomedical Engineering, Yale University, New Haven, CT.
authors.forename: W Mark
authors.initials: WM
authors.lastname: Saltzman
authors.affiliation: Department of Comparative Medicine, Yale School of Medicine, New Haven, CT.
authors.forename: Caroline J
authors.initials: CJ
authors.lastname: Zeiss
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT.
authors.forename: Floriana
authors.initials: F
authors.lastname: Centritto
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT.
authors.forename: Jonathan D
authors.initials: JD
authors.lastname: Black
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT.
authors.forename: Dan-Arin
authors.initials: DA
authors.lastname: Silasi
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT.
authors.forename: Elena
authors.initials: E
authors.lastname: Ratner
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT.
authors.forename: Masoud
authors.initials: M
authors.lastname: Azodi
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Rutherford
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT.
authors.forename: Peter E
authors.initials: PE
authors.lastname: Schwartz
authors.affiliation: Division of Gynecologic Oncology, University of Brescia, Brescia, Italy.
authors.forename: Sergio
authors.initials: S
authors.lastname: Pecorelli
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale School of Medicine, New Haven, CT.
authors.forename: Alessandro D
authors.initials: AD
authors.lastname: Santin
mesh_terms: D000818:Animals; D062465:Claudin-3 / Q000378:metabolism; D062506:Claudin-4 / Q000378:metabolism; D004768:Enterotoxins / Q000008:administration & dosage*; D005260:Female; D005456:Fluorescent Dyes / Q000008:administration & dosage*; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D061206:Neoplasm Micrometastasis / Q000473:pathology*; D010051:Ovarian Neoplasms / Q000473:pathology*; D023041:Xenograft Model Antitumor Assays / Q000379:methods
chemical_list: D062465:Claudin-3; D062506:Claudin-4; D004768:Enterotoxins; D005456:Fluorescent Dyes; C035598:enterotoxin, Clostridium
keywords: Clostridium perfringens enterotoxin; IRDye CW800; ovarian cancer; real-time imaging; residual disease