In Vitro Evaluation of Radiolabeled Amphotericin B for Molecular Imaging of Mold Infections

Abstract

Invasive pulmonary aspergillosis and mucormycosis are life-threatening complications in immunocompromised patients. A rapid diagnosis followed by early antifungal treatment is essential for patient survival. Given the limited spectrum of biomarkers for invasive mold infections, recent studies have proposed the use of radiolabeled siderophores or antibodies as molecular probes to increase the specificity of radiological findings by nuclear imaging modalities. While holding enormous di-agnostic potential, most of the currently available molecular probes are tailored to the detection of Aspergillus species, and their cost-intensive and sophisticated imple-mentation restricts their accessibility at less specialized centers. In order to develop cost-efficient and broadly applicable tracers for pulmonary mold infections, this study established streamlined and high-yielding protocols to radiolabel amphotericin B (AMB) with the gamma emitter technetium-99m ((99)m Tc-AMB) and the positron emitter gallium-68 (Ga-68-AMB). The radiochemical purity of the resulting tracers con-sistently exceeded 99%, and both probes displayed excellent stability in human se-rum (98% after 60 to 240 min at 37 degrees C). The uptake kinetics by representative mold pathogens were assessed in an in vitro Transwell assay using infected endothelial cell layers. Both tracers accumulated intensively and specifically in Transwell inserts infected with Aspergillus fumigatus , Rhizopus arrhizus , and other clinically relevant mold pathogens compared with their accumulation in uninfected inserts and inserts infected with bacterial controls. Inoculum-dependent enrichment was confirmed by gamma counting and autoradiographic imaging. Taken together, this pilot in vitro study proposes Tc-99m-AMB and Ga-68-AMB to be facile, stable, and specific probes, meriting further preclinical in vivo evaluation of radiolabeled amphoteri-cin B for molecular imaging in invasive mycoses.