Molecular probes for targeting facilitative fructose transporters (GLUTs) in biochemical and biomedical applications

The primary goal is to advance the development of GLUT-targeting molecular probes to facilitate the characterization of cells and discrimination between cancer and normal cells. High sugar uptake has long been identified as a hallmark of cancer. Recently, the specific dependence of cancers on fructose for development and progression outlined fructose uptake as a potential diagnostic and therapeutic target. Respectively, changes in the activity of facilitative fructose transporters (GLUTs) and alterations in the overall GLUT profile between normal and metabolically compromised cells have set the foundation for developing GLUT-based approaches to target cancer cells. However, specific targeting of disease-relevant fructose GLUTs remains a challenge, limiting the use of GLUTs in diagnostic and therapeutic applications. The proposed research program demonstrates the targeting of selected fructose GLUTs (GLUT2, GLUT12, and GLUT5) for identifying fructose-dependent cells in imaging applications. The proposal addresses several challenges in developing GLUT-specific molecular probes, GLUT-targeting cell imaging, and GLUT activity screening approaches. The primary innovative component enables discrimination between nonspecific fructose GLUTs allowing for designing specific molecular probes for several cancer-relevant fructose transporters. The developments proposed in this research include approaches for GLUT-based cell profiling, GLUT-based characterization of cancer cells, and GLUTbased in vivo imaging. The direct outcomes of this research include the validation of a novel multi-color multiGLUT screening approach for measuring GLUT5:GLUT2 activity ratios through fluorescence and novel turn-on fluorescent molecular probes for real-time monitoring and high-throughput fluorescence screening of GLUT activity. The project will yield a new GLUT-based characterization of different breast cancer cell lines and identify the fructose-dependent cell lines potentially targeted for in vivo imaging by GLUT-specific probes. The project also advances the specific targeting of fructose GLUTs in vivo by developing GLUT-specific imaging probes for PET applications. The research project is designed to be carried out primarily by undergraduate students under the supervision of the PI and a graduate researcher. The program provides a comprehensive multidisciplinary educational platform for undergraduate and graduate researchers and offers a hands-on research experience at the interface of chemistry and biology. By engineering probes that can specifically target other fructose or glucose transporters, we will advance to fingerprinting GLUTs to characterize and distinguish cells. GLUT-specific probes will enable fundamental studies to foster an understanding of the impact of each GLUT and different dietary sugars at the molecular level on health and disease.