br Statistics br Each examination was performed in
Each examination was performed in triplicate. Statistical differences between clinicopathologic parameters and the miR-143 level of tumor samples were evaluated by using Pearson’s c2 test or Fisher’s extract test, unless otherwise specified. For in vitro and in vivo experiments, statistical significances of differences were evaluated by performing the two-sided Student’s t test. The values were presented as the mean ± SD. A p value < 0.05 was considered to be statistically signif-icant. The effects of intravesical injection of miR-143#12 on animal survival were evaluated by preparing Kaplan-Meier plots, with the survival duration analyzed for significance by performing log-rank survival analysis.
CONFLICTS OF INTEREST
The authors declare no conflicts of interest.
This work was supported by the Project for Cancer Research and Therapeutic Evolution (P-CREATE) from Japan Agency for Medical Research and Development (AMED 16cm 0106202h0001). This work was also performed with the support of SHIONOGI. We are grateful for strong technical support by Nobuhiko Sugito, Yuki Kuranaga, Minami Kumazaki, and Haruko Shinohara (United Graduate School of Drug Discovery and Medical Information Sciences). We also thank Drs. Taizo Uchimoto, Kenkichi Saito, and Naoki Tanda (Department of Urology, Osaka Medical College) for their help with sample acquisition.
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Molecular Therapy: Methods & Clinical Development
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Association for Academic Surgery
Anti-Claudin-1 Conjugated to a Near-Infrared Fluorophore Targets Colon Cancer in PDOX Mouse Models
Hannah M. Hollandsworth, MD,a,b,c Thinzar M. Lwin, MD,a,b,c Siamak Amirfakhri, PhD,a,b,c Filemoni Filemoni, a,b,c,d Surinder K. Batra, PhD,e Robert M. Hoffman, PhD,a,b,c,d Punita Dhawan, PhD,e and Michael Bouvet, MDa,b,c,*
a Department of Surgery, University of California San Diego, La Jolla, California
b Moores Cancer Center, University of California San Diego, La Jolla, California
c Department of Surgery, VA San Diego Healthcare System, San Diego, California
d AntiCancer, Inc, San Diego, California
e Department of Biochemistry, University of Nebraska Medical Center, Omaha, Nebraska
Near infrared dye
Fluorescence guided surgery
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. An-imal 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 adminis-tered 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 mg of Claudin-1-IRDye800CW.