Clostridium perfringens enterotoxin C-terminal domain labeled to fluorescent dyes for in vivo visualization of micrometastatic chemotherapy-resistant ovarian cancer

Emiliano Cocco, Erik M. Shapiro, Sara Gasparrini, Salvatore Lopez, Carlton L. Schwab, Stefania Bellone, Ileana Bortolomai, Natalia J. Sumi, Elena Bonazzoli, Roberta Nicoletti, Yang Deng, W. Mark Saltzman, Caroline J. Zeiss, Floriana Centritto, Jonathan D. Black, Dan Arin Silasi, Elena Ratner, Masoud Azodi, Thomas J. Rutherford, Peter E. Schwartz & 2 others Sergio Pecorelli, Alessandro D. Santin

    Research output: Research - peer-reviewArticle

    • 12 Citations

    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. What's new? Intraoperative detection of residual tumor is an emerging technique aimed at improving the prognosis of women with ovarian cancer. The authors developed a new system using the carboxy-terminal fragment of Clostridium Perfringens Enterotoxin (c-CPE) conjugated to fluorescent fluorophores to successfully mark microscopic tumor deposits in mouse models of chemotherapy-naïve and chemotherapy-resistant ovarian cancer. C-CPE binds to claudin 3/4 receptors, which the authors showed earlier are highly overexpressed on most types of ovarian cancer, thus providing a new tumor-specific tool to minimize micrometastic disease in patients with ovarian cancer.

    LanguageEnglish (US)
    Pages2618-2629
    Number of pages12
    JournalInternational Journal of Cancer
    Volume137
    Issue number11
    DOIs
    StatePublished - Dec 1 2015

    Profile

    Fluorescent Dyes
    Ovarian Neoplasms
    Drug Therapy
    Clostridium enterotoxin
    Neoplasms
    Fluorescence
    Claudin-3
    Claudin-4
    Fluorescein-5-isothiocyanate
    Ascites
    Heterografts
    Peptides
    Injections
    Ovarian Diseases
    Neoadjuvant Therapy
    Residual Neoplasm
    Fluorescence Microscopy
    Observation

    Keywords

    • Clostridium perfringens enterotoxin
    • IRDye CW800
    • ovarian cancer
    • real-time imaging
    • residual disease

    ASJC Scopus subject areas

    • Cancer Research
    • Oncology

    Cite this

    Clostridium perfringens enterotoxin C-terminal domain labeled to fluorescent dyes for in vivo visualization of micrometastatic chemotherapy-resistant ovarian cancer. / Cocco, Emiliano; Shapiro, Erik M.; Gasparrini, Sara; Lopez, Salvatore; Schwab, Carlton L.; Bellone, Stefania; Bortolomai, Ileana; Sumi, Natalia J.; Bonazzoli, Elena; Nicoletti, Roberta; Deng, Yang; Saltzman, W. Mark; Zeiss, Caroline J.; Centritto, Floriana; Black, Jonathan D.; Silasi, Dan Arin; Ratner, Elena; Azodi, Masoud; Rutherford, Thomas J.; Schwartz, Peter E.; Pecorelli, Sergio; Santin, Alessandro D.

    In: International Journal of Cancer, Vol. 137, No. 11, 01.12.2015, p. 2618-2629.

    Research output: Research - peer-reviewArticle

    Cocco, E, Shapiro, EM, Gasparrini, S, Lopez, S, Schwab, CL, Bellone, S, Bortolomai, I, Sumi, NJ, Bonazzoli, E, Nicoletti, R, Deng, Y, Saltzman, WM, Zeiss, CJ, Centritto, F, Black, JD, Silasi, DA, Ratner, E, Azodi, M, Rutherford, TJ, Schwartz, PE, Pecorelli, S & Santin, AD 2015, 'Clostridium perfringens enterotoxin C-terminal domain labeled to fluorescent dyes for in vivo visualization of micrometastatic chemotherapy-resistant ovarian cancer' International Journal of Cancer, vol 137, no. 11, pp. 2618-2629. DOI: 10.1002/ijc.29632
    Cocco, Emiliano ; Shapiro, Erik M. ; Gasparrini, Sara ; Lopez, Salvatore ; Schwab, Carlton L. ; Bellone, Stefania ; Bortolomai, Ileana ; Sumi, Natalia J. ; Bonazzoli, Elena ; Nicoletti, Roberta ; Deng, Yang ; Saltzman, W. Mark ; Zeiss, Caroline J. ; Centritto, Floriana ; Black, Jonathan D. ; Silasi, Dan Arin ; Ratner, Elena ; Azodi, Masoud ; Rutherford, Thomas J. ; Schwartz, Peter E. ; Pecorelli, Sergio ; Santin, Alessandro D./ Clostridium perfringens enterotoxin C-terminal domain labeled to fluorescent dyes for in vivo visualization of micrometastatic chemotherapy-resistant ovarian cancer. In: International Journal of Cancer. 2015 ; Vol. 137, No. 11. pp. 2618-2629
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    AU - Shapiro,Erik M.

    AU - Gasparrini,Sara

    AU - Lopez,Salvatore

    AU - Schwab,Carlton L.

    AU - Bellone,Stefania

    AU - Bortolomai,Ileana

    AU - Sumi,Natalia J.

    AU - Bonazzoli,Elena

    AU - Nicoletti,Roberta

    AU - Deng,Yang

    AU - Saltzman,W. Mark

    AU - Zeiss,Caroline J.

    AU - Centritto,Floriana

    AU - Black,Jonathan D.

    AU - Silasi,Dan Arin

    AU - Ratner,Elena

    AU - Azodi,Masoud

    AU - Rutherford,Thomas J.

    AU - Schwartz,Peter E.

    AU - Pecorelli,Sergio

    AU - Santin,Alessandro D.

    PY - 2015/12/1

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    N2 - 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. What's new? Intraoperative detection of residual tumor is an emerging technique aimed at improving the prognosis of women with ovarian cancer. The authors developed a new system using the carboxy-terminal fragment of Clostridium Perfringens Enterotoxin (c-CPE) conjugated to fluorescent fluorophores to successfully mark microscopic tumor deposits in mouse models of chemotherapy-naïve and chemotherapy-resistant ovarian cancer. C-CPE binds to claudin 3/4 receptors, which the authors showed earlier are highly overexpressed on most types of ovarian cancer, thus providing a new tumor-specific tool to minimize micrometastic disease in patients with ovarian cancer.

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    KW - real-time imaging

    KW - residual disease

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