26.04 Fluorophore Conjugated Claudin-1 Antibodies Improve Detection of Colon Polyps in CPC-APC Mice

S. Liu1, K. Cox1, S. Amirfakhri1, S. Jaiswal1, R. Hoffman1, S. Batra2, M. Primeaux2, P. Dhawan2, S. Talib2, A. Mohs2, M. Bouvet1  1University of California, San Diego, Surgery, San Diego, CA, USA 2University Of Nebraska College Of Medicine, Omaha, NE, USA

Introduction:

Early detection and removal of pre-malignant polyps during colonoscopy remains the best way to prevent colon cancer. Colonoscopy is technically challenging and can have miss rates between 9% – 27% for pre-malignant polyps up to 1 cm in diameter. Additionally, hyperplastic polyps (HP) have little to no malignant potential but can be indistinguishable from pre-malignant sessile serrated adenomas (SSA) on visual exam. This study uses Claudin-1 antibodies conjugated to near-infrared fluorophores to increase detection of precancerous polyps in adenomatous polyposis coli flox mice with a Cdx2-Cre transgene (CPC-APC) that develop colonic polyps with the goal of translation to human use.

Methods:

CPC-APC mice at age 12 weeks at which time colonic polyps have developed were used for this study. Claudin-1 antibody conjugated to IRDye800 (Claudin-1-IR800) was administered intravenously at a dose of 100 ug. Mice were sacrificed post injection at 48 and 72 hour timepoints. Ex-vivo colon imaging was performed using the Pearl and the Stryker 1688 fluorescent imaging systems. Immunohistochemistry of the polyps was performed to confirm Claudin-1 expression. Polyp to background fluroescence signal ratios (PBR) were calculated.

Results:

Four mice were used yielding a total of 15 polyps that were analyzed. The presence of observed polyps on gross examination was confirmed via histology. The PBR at 48 hours post injection was 6.8 ± 3.18 and at 72 hours it was 7.95 ± 0.78 (P = 0.093). Overall, the average PBR was 6.81 ± 2.9, resulting in an over 6-fold fluorescence signal intensity at the site of polyps. Immunohistochemistry confirmed strong expression of Claudin-1 within the polyps. The Stryker 1688 laparoscope was used on the gross colon specimens to confirm visibility of polyps using a commercially available clinical fluorescence imaging system. The fluorescence signal could be definitively detected in polyps as small as 1mm in mice that were invisible in bright light (see Figure). 

Conclusion:

Claudin-1-IR800 demonstrated high sensitivity for polyp identification in the CPC-APC mice with an average PBR of >6. Polyps could be detected as small as 1 mm that were invisible in bright light, greatly improving the visual accuracy in polyp identification. Future studies are planned that translate the present findings to clinical practice.