PURPOSE: To assess the potential of αvβ6-integrin as a theranostic target in esophageal cancer. METHODS: Membranous β6-integrin (ITGB6) expression was analyzed in 306 specimens of human esophageal squamous cell carcinoma (ESCC) obtained by immunohistochemistry (IHC) from 100 patient cases (1, 37, 58, and 4 of grade G1, G2, G3, and G4, respectively). Ga-68 labeling of D0103 was done manually for preclinical experiments and fully automated for clinical application. Preclinical characterization of Ga-68-D0103 was performed in SCID mice bearing subcutaneous xenografts of H2009 (αvβ6-positive) or MDA-MB-231 (αvβ6-negative) carcinoma cell lines, by ex vivo biodistribution (10, 30, 90, and 180 min p.i) and PET imaging (30, 90, and 180 min p.i.)., without and with co-injection of gelofusine (4% succinylated gelatin). A patient with type-II diabetes (f, 68y, 115 kg) with proximal G2 ESCC was investigated by Ga-68-D0103 PET/CT (193 MBq) at 15, 45, 90, and 104 min p.i.. RESULTS: 99% of ESCC cases were found β6-integrin positive by IHC, of which 48%, 31%, and 20% showed strong, moderate, and low ITGB6 expression, respectively, with no correlation to tumor grade. Ex vivo biodistribution of Ga-68-D0103 in H2009 xenografted mice after 30, 90, and 180 min showed tumor-to-blood ratios of 6.8, 37, and 124, respectively; tumor-to-muscle ratios of 12, 14, and 36, respectively; tumor-to-liver ratios of 10, 17, and 14, respectively; and tumor-to-pancreas ratios of 20, 47, and 56, respectively. Co-administration of gelofusine did not change the tumor uptake but reduced the kidney uptake by 89% (from 178%iA/g to 19.1%iA/g, 90 min p.i.), resulting in an 8.7-fold higher tumor/kidney ratio. µPET imaging in H2009 xenografted mice confirmed a high tumor uptake and low background already 30 min p.i.. Blockade biodistribution and µPET in αvβ6-(-) MDA-MB-231 mice demonstrated target specificity. Clinical PET/CT of a patient with ESCC showed increasing tracer uptake over time in the primary tumor (SUVmax 9.0 and 11.3 at 15 and 104 min p.i., respectively) and in a lymph node metastasis (SUVmax 19.5 and 28.3, respectively), and a decreasing blood pool activity (SUVmean 2.75 and 0.98, respectively). CONCLUSIONS: High (99%) membranous expression frequency and density on tumor cells underscores the potential of αvβ6-integrin as a theranostic target in ESCC, suggesting that αvβ6-integrin PET/CT imaging may adopt a role in re-staging and therapy guidance in this cancer type. The prolonged tumor retention furthermore indicates a therapeutic potential of αvβ6-integrin targeted radiopharmaceuticals when labeled with radionuclides such as lutetium-177, terbium-161, or actinium-225.

Czech Advanced Technology and Research Institute Palacký University Olomouc Olomouc Czech Republic

Division of Nuclear Medicine Department of Radiology and Nuclear Medicine University Hospital Magdeburg Magdeburg Germany

Institute of Molecular and Translational Medicine Faculty of Medicine and Dentistry Palacký University Olomouc Olomouc Czech Republic

Institute of Pathology School of Medicine and Health Technical University of Munich Munich Germany

Institute of Pathology University Hospital Marburg 35043 Marburg Germany

Laboratory of Experimental Medicine Institute of Molecular and Translational Medicine University Hospital Olomouc Olomouc Czech Republic

TRIMT GmbH Carl Eschebach Str 7 01454 Radeberg Germany

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