Anti-Claudin-1 Conjugated to a Near-Infrared Fluorophore Targets Colon Cancer in PDOX Mouse Models

Hannah M. Hollandsworth, Thinzar M. Lwin, Siamak Amirfakhri, Filemoni Filemoni, Surinder Kumar Batra, Robert M. Hoffman, Punita Dhawan, Michael Bouvet

Research output: Contribution to journalArticle

Abstract

Introduction: Claudins are tight-junction proteins, which maintain an epithelial barrier in normal colon cells. Overexpression of Claudin-1 has been implicated for development of colon cancer. We postulated that Claudin-1 may be a useful target in near-infrared imaging and fluorescence-guided surgery. Methods: We conjugated Claudin-1 antibody to LI-COR IR800DyeCW (Claudin-1-IRDye800CW). Western blotting of 9 human colon cancer cell lysates was performed. Animal imaging was performed with the LI-COR Pearl Trilogy Fluorescence Imaging System. A dose-response study was carried out with subcutaneous LS174T colon cancer cell line models. Increasing doses of Claudin-1-IRDye800CW via tail vein injection were administered to three groups of mice. Two groups of mice were used as controls (antibody alone, and dye alone). In vivo imaging was performed at 24, 48, and 72 h after administration of the conjugated dye. Orthotopic implantation of patient-derived tumors and cell lines was performed and peritoneal carcinomatosis models were created. After tumor growth, mice were administered Claudin-1-IRDye800CW and imaged in vivo 48 h later. The mice were euthanized and laparotomy was performed to assess internal organs and toxicity. Results: Western blotting revealed that all colon cancer cell lysates expressed varying amounts of Claudin-1. All tumors demonstrated strong and specific fluorescence labeling at 800 nm, even with the lowest dose of 12.5 μg of Claudin-1-IRDye800CW. Conclusions: Claudin-1 is a useful target for near-infrared antibody-based imaging for visualization of colorectal tumors for future use in fluorescence-guided surgery.

Original languageEnglish (US)
Pages (from-to)145-150
Number of pages6
JournalJournal of Surgical Research
Volume242
DOIs
StatePublished - Oct 1 2019

Fingerprint

Claudin-1
Colonic Neoplasms
Optical Imaging
Antibodies
Coloring Agents
Fluorescence
Western Blotting
Claudins
Tight Junction Proteins
Tumor Cell Line
Laparotomy
Tail
Colorectal Neoplasms
Veins
Neoplasms
Colon
Carcinoma

Keywords

  • Claudin
  • Colon cancer
  • Fluorescence guided surgery
  • IRDye800CW
  • Near infrared dye

ASJC Scopus subject areas

  • Surgery

Cite this

Hollandsworth, H. M., Lwin, T. M., Amirfakhri, S., Filemoni, F., Batra, S. K., Hoffman, R. M., ... Bouvet, M. (2019). Anti-Claudin-1 Conjugated to a Near-Infrared Fluorophore Targets Colon Cancer in PDOX Mouse Models. Journal of Surgical Research, 242, 145-150. https://doi.org/10.1016/j.jss.2019.04.048

Anti-Claudin-1 Conjugated to a Near-Infrared Fluorophore Targets Colon Cancer in PDOX Mouse Models. / Hollandsworth, Hannah M.; Lwin, Thinzar M.; Amirfakhri, Siamak; Filemoni, Filemoni; Batra, Surinder Kumar; Hoffman, Robert M.; Dhawan, Punita; Bouvet, Michael.

In: Journal of Surgical Research, Vol. 242, 01.10.2019, p. 145-150.

Research output: Contribution to journalArticle

Hollandsworth, Hannah M. ; Lwin, Thinzar M. ; Amirfakhri, Siamak ; Filemoni, Filemoni ; Batra, Surinder Kumar ; Hoffman, Robert M. ; Dhawan, Punita ; Bouvet, Michael. / Anti-Claudin-1 Conjugated to a Near-Infrared Fluorophore Targets Colon Cancer in PDOX Mouse Models. In: Journal of Surgical Research. 2019 ; Vol. 242. pp. 145-150.
@article{fc4e680a234348edb8e3a08e5f9b915f,
title = "Anti-Claudin-1 Conjugated to a Near-Infrared Fluorophore Targets Colon Cancer in PDOX Mouse Models",
abstract = "Introduction: Claudins are tight-junction proteins, which maintain an epithelial barrier in normal colon cells. Overexpression of Claudin-1 has been implicated for development of colon cancer. We postulated that Claudin-1 may be a useful target in near-infrared imaging and fluorescence-guided surgery. Methods: We conjugated Claudin-1 antibody to LI-COR IR800DyeCW (Claudin-1-IRDye800CW). Western blotting of 9 human colon cancer cell lysates was performed. Animal imaging was performed with the LI-COR Pearl Trilogy Fluorescence Imaging System. A dose-response study was carried out with subcutaneous LS174T colon cancer cell line models. Increasing doses of Claudin-1-IRDye800CW via tail vein injection were administered to three groups of mice. Two groups of mice were used as controls (antibody alone, and dye alone). In vivo imaging was performed at 24, 48, and 72 h after administration of the conjugated dye. Orthotopic implantation of patient-derived tumors and cell lines was performed and peritoneal carcinomatosis models were created. After tumor growth, mice were administered Claudin-1-IRDye800CW and imaged in vivo 48 h later. The mice were euthanized and laparotomy was performed to assess internal organs and toxicity. Results: Western blotting revealed that all colon cancer cell lysates expressed varying amounts of Claudin-1. All tumors demonstrated strong and specific fluorescence labeling at 800 nm, even with the lowest dose of 12.5 μg of Claudin-1-IRDye800CW. Conclusions: Claudin-1 is a useful target for near-infrared antibody-based imaging for visualization of colorectal tumors for future use in fluorescence-guided surgery.",
keywords = "Claudin, Colon cancer, Fluorescence guided surgery, IRDye800CW, Near infrared dye",
author = "Hollandsworth, {Hannah M.} and Lwin, {Thinzar M.} and Siamak Amirfakhri and Filemoni Filemoni and Batra, {Surinder Kumar} and Hoffman, {Robert M.} and Punita Dhawan and Michael Bouvet",
year = "2019",
month = "10",
day = "1",
doi = "10.1016/j.jss.2019.04.048",
language = "English (US)",
volume = "242",
pages = "145--150",
journal = "Journal of Surgical Research",
issn = "0022-4804",
publisher = "Academic Press Inc.",

}

TY - JOUR

T1 - Anti-Claudin-1 Conjugated to a Near-Infrared Fluorophore Targets Colon Cancer in PDOX Mouse Models

AU - Hollandsworth, Hannah M.

AU - Lwin, Thinzar M.

AU - Amirfakhri, Siamak

AU - Filemoni, Filemoni

AU - Batra, Surinder Kumar

AU - Hoffman, Robert M.

AU - Dhawan, Punita

AU - Bouvet, Michael

PY - 2019/10/1

Y1 - 2019/10/1

N2 - Introduction: Claudins are tight-junction proteins, which maintain an epithelial barrier in normal colon cells. Overexpression of Claudin-1 has been implicated for development of colon cancer. We postulated that Claudin-1 may be a useful target in near-infrared imaging and fluorescence-guided surgery. Methods: We conjugated Claudin-1 antibody to LI-COR IR800DyeCW (Claudin-1-IRDye800CW). Western blotting of 9 human colon cancer cell lysates was performed. Animal imaging was performed with the LI-COR Pearl Trilogy Fluorescence Imaging System. A dose-response study was carried out with subcutaneous LS174T colon cancer cell line models. Increasing doses of Claudin-1-IRDye800CW via tail vein injection were administered to three groups of mice. Two groups of mice were used as controls (antibody alone, and dye alone). In vivo imaging was performed at 24, 48, and 72 h after administration of the conjugated dye. Orthotopic implantation of patient-derived tumors and cell lines was performed and peritoneal carcinomatosis models were created. After tumor growth, mice were administered Claudin-1-IRDye800CW and imaged in vivo 48 h later. The mice were euthanized and laparotomy was performed to assess internal organs and toxicity. Results: Western blotting revealed that all colon cancer cell lysates expressed varying amounts of Claudin-1. All tumors demonstrated strong and specific fluorescence labeling at 800 nm, even with the lowest dose of 12.5 μg of Claudin-1-IRDye800CW. Conclusions: Claudin-1 is a useful target for near-infrared antibody-based imaging for visualization of colorectal tumors for future use in fluorescence-guided surgery.

AB - Introduction: Claudins are tight-junction proteins, which maintain an epithelial barrier in normal colon cells. Overexpression of Claudin-1 has been implicated for development of colon cancer. We postulated that Claudin-1 may be a useful target in near-infrared imaging and fluorescence-guided surgery. Methods: We conjugated Claudin-1 antibody to LI-COR IR800DyeCW (Claudin-1-IRDye800CW). Western blotting of 9 human colon cancer cell lysates was performed. Animal imaging was performed with the LI-COR Pearl Trilogy Fluorescence Imaging System. A dose-response study was carried out with subcutaneous LS174T colon cancer cell line models. Increasing doses of Claudin-1-IRDye800CW via tail vein injection were administered to three groups of mice. Two groups of mice were used as controls (antibody alone, and dye alone). In vivo imaging was performed at 24, 48, and 72 h after administration of the conjugated dye. Orthotopic implantation of patient-derived tumors and cell lines was performed and peritoneal carcinomatosis models were created. After tumor growth, mice were administered Claudin-1-IRDye800CW and imaged in vivo 48 h later. The mice were euthanized and laparotomy was performed to assess internal organs and toxicity. Results: Western blotting revealed that all colon cancer cell lysates expressed varying amounts of Claudin-1. All tumors demonstrated strong and specific fluorescence labeling at 800 nm, even with the lowest dose of 12.5 μg of Claudin-1-IRDye800CW. Conclusions: Claudin-1 is a useful target for near-infrared antibody-based imaging for visualization of colorectal tumors for future use in fluorescence-guided surgery.

KW - Claudin

KW - Colon cancer

KW - Fluorescence guided surgery

KW - IRDye800CW

KW - Near infrared dye

UR - http://www.scopus.com/inward/record.url?scp=85065164577&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85065164577&partnerID=8YFLogxK

U2 - 10.1016/j.jss.2019.04.048

DO - 10.1016/j.jss.2019.04.048

M3 - Article

VL - 242

SP - 145

EP - 150

JO - Journal of Surgical Research

JF - Journal of Surgical Research

SN - 0022-4804

ER -