ABSTRACT
The treatment of choice for insulinomas and focal lesions in congenital hyperinsulinism (CHI) is surgery. However, intraoperative detection can be challenging. This challenge could be overcome with intraoperative fluorescence imaging, which provides real-time lesion detection with a high spatial resolution. Here, a novel method for targeted near-infrared (NIR) fluorescence imaging of glucagonlike peptide 1 receptor (GLP-1R)-positive lesions, using the GLP-1 agonist exendin-4 labeled with IRDye 800CW, was examined in vitro and in vivo. Methods: A competitive binding assay was performed using Chinese hamster lung (CHL) cells transfected with GLP-1R. Tracer biodistribution was determined in BALB/c nude mice bearing subcutaneous CHL-GLP-1R xenografts. In vivo NIR fluorescence imaging of CHL-GLP-1R xenografts was performed. Localization of the tracer in the pancreatic islets of BALB/c nude mice was examined using fluorescence microscopy. Laparoscopic imaging was performed to detect the fluorescent signal of the tracer in the pancreas of mini pigs. Results: Exendin-4-IRDye 800CW binds GLP-1R with a half-maximal inhibitory concentration of 3.96 nM. The tracer accumulates in CHL-GLP-1R xenografts. Subcutaneous CHL-GLP-1R xenografts were visualized using in vivo NIR fluorescence imaging. The tracer accumulates specifically in the pancreatic islets of mice, and a clear fluorescent signal was detected in the pancreas of mini pigs. Conclusion: These data provide the first in vivo evidence of the feasibility of targeted fluorescence imaging of GLP-1R-positive lesions. Intraoperative lesion delineation using exendin-4-IRDye 800CW could benefit open as well as laparoscopic surgical procedures for removal of insulinomas and focal lesions in CHI.
Subject(s)
Benzenesulfonates/chemistry , Exenatide/chemistry , Glucagon-Like Peptide-1 Receptor/metabolism , Indoles/chemistry , Optical Imaging/methods , Animals , Biological Transport , CHO Cells , Cricetulus , Exenatide/metabolism , Exenatide/pharmacokinetics , Female , Mice , Mice, Nude , Pancreas/metabolism , Swine , Tissue DistributionABSTRACT
The identification of a beta-cell tracer is a major quest in diabetes research. However, since MRI, PET and SPECT cannot resolve individual islets, optical techniques are required to assess the specificity of these tracers. We propose to combine Optical Coherence Microscopy (OCM) with fluorescence detection in a single optical platform to facilitate these initial screening steps from cell culture up to living rodents. OCM can image islets and vascularization without any labeling. Thereby, it alleviates the need of both genetically modified mice to detect islets and injection of external dye to reveal vascularization. We characterized Cy5.5-exendin-3, an agonist of glucagon-like peptide 1 receptor (GLP1R), for which other imaging modalities have been used and can serve as a reference. Cultured cells transfected with GLP1R and incubated with Cy5.5-exendin-3 show full tracer internalization. We determined that a dose of 1 µg of Cy5.5-exendin-3 is sufficient to optically detect in vivo the tracer in islets with a high specificity. In a next step, time-lapse OCM imaging was used to monitor the rapid and specific tracer accumulation in murine islets and its persistence over hours. This optical platform represents a versatile toolbox for selecting beta-cell specific markers for diabetes research and future clinical diagnosis.
Subject(s)
Carbocyanines/pharmacology , Diabetes Mellitus/metabolism , Glucagon-Like Peptide-1 Receptor/analysis , Insulin-Secreting Cells/cytology , Tomography, Optical Coherence/methods , Animals , Cell Line , Cricetulus , Female , Fluorescent Dyes , Glucagon-Like Peptide-1 Receptor/agonists , Glucagon-Like Peptide-1 Receptor/genetics , Mice , Mice, Inbred ICR , PeptidesABSTRACT
The evaporation rate of water is, of course, different under the same heating conditions of different aqueous solutions. Under conventional heating conditions, the evaporation rate of water is much higher than the evaporation rate of water of aqueous solutions of different kinds of solute materials, which is well accordance with the classical Raoult's law. The results obtained in this study have clearly shown that the chemical characteristics of dissolved materials in water very seriously affect the evaporation rates of water under the microwave heating. This generally causes contradictory results to Raoult's law and this can be explained with the additional microwave energy absorption by the ionic or molecular solute materials found in the solutions other than the microwave energy absorption by water molecules themselves.
