Diagnostic PET Imaging of Mammary Microcalcifications Using ⁶⁴Cu-DOTA-Alendronate in a Rat Model of Breast Cancer.

Abstract

The development of improved breast cancer screening methods is hindered by a lack of cancer-specific imaging agents and effective small-animal models to test them. The purpose of this study was to evaluate ⁶⁴Cu-DOTA-alendronate as a mammary microcalcification-targeting PET imaging agent, using an ideal rat model. Our long-term goal is to develop ⁶⁴Cu-DOTA-alendronate for the detection and noninvasive differentiation of malignant versus benign breast tumors with PET. <b>Methods:</b> DOTA-alendronate was synthesized, radiolabeled with ⁶⁴Cu, and administered to normal or tumor-bearing aged, female, retired breeder Sprague-Dawley rats for PET imaging. Mammary tissues were subsequently labeled and imaged with light, confocal, and electron microscopy to verify microcalcification targeting specificity of DOTA-alendronate and elucidate the histologic and ultrastructural characteristics of the microcalcifications in different mammary tumor types. Tumor uptake, biodistribution, and dosimetry studies were performed to evaluate the efficacy and safety of ⁶⁴Cu-DOTA-alendronate. <b>Results:</b>⁶⁴Cu-DOTA-alendronate was radiolabeled with a 98% yield. PET imaging using aged, female, retired breeder rats showed specific binding of ⁶⁴Cu-DOTA-alendronate in mammary glands and mammary tumors. The highest uptake of ⁶⁴Cu-DOTA-alendronate was in malignant tumors and the lowest uptake in benign tumors and normal mammary tissue. Confocal analysis with carboxyfluorescein-alendronate confirmed the microcalcification binding specificity of alendronate derivatives. Biodistribution studies revealed tissue alendronate concentrations peaking within the first hour, then decreasing over the next 48 h. Our dosimetric analysis demonstrated a ⁶⁴Cu effective dose within the acceptable range for clinical PET imaging agents and the potential for translation into human patients. <b>Conclusion:</b>⁶⁴Cu-DOTA-alendronate is a promising PET imaging agent for the sensitive and specific detection of mammary tumors as well as the differentiation of malignant versus benign tumors based on absolute labeling uptake.