Metabolism-Driven High-Throughput Cancer Identification with GLUT5-Specific Molecular Probes

Abstract

Point-of-care applications rely on biomedical sensors to enable rapid detection with high sensitivity and selectivity. Despite advances in sensor development, there are challenges in cancer diagnostics. Detection of biomarkers, cell receptors, circulating tumor cells, gene identification, and fluorescent tagging are time-consuming due to the sample preparation and response time involved. Here, we present a novel approach to target the enhanced metabolism in breast cancers for rapid detection using fluorescent imaging. Fluorescent analogs of fructose target the fructose-specific transporter GLUT5 in breast cancers and have limited to no response from normal cells. These analogs demonstrate a marked difference in adenocarcinoma and premalignant cells leading to a novel detection approach. The vastly different uptake kinetics of the analogs yields two unique signatures for each cell type. We used normal breast cells MCF10A, adenocarcinoma cells MCF7, and premalignant cells MCF10AneoT, with hepatocellular carcinoma cells HepG2 as the negative control. Our data indicated that MCF10AneoT and MCF7 cells had an observable difference in response to only one of the analogs. The response, observed as fluorescence intensity, leads to a two-point assessment of the cells in any sample. Since the treatment time is 10 min, there is potential for use in rapid on-site high-throughput diagnostics.

Targeting of GLUT5 for Transporter-Mediated Drug-Delivery Is Contingent upon Substrate Hydrophilicity

“…GLUT5-targeting conjugates were constructed from the 2,5-anhydro-D-mannitol (Man) as the locked fructofuranose mimic and various coumarins (Cou) as fluorescent moieties. The resulting ManCou probes (ManCou-CH 3 representative in Figure 1 ) were found useful in specifically targeting GLUT5 for imaging applications and transport activity evaluation in vitro in the complete cell culture media, enabling live cell analysis [ 22 , 23 ]. Following this attainment, we contemplated assessing the tolerance of fructose transporter GLUT5 towards passing a bioactive moiety in a glycoconjugate form and the feasibility of achieving cancer-specific cytotoxicity.…”

Insights into Substrate and Inhibitor Selectivity among Human GLUT Transporters through Comparative Modeling and Molecular Docking

“…Toward this goal, inhibitors selective for either GLUT1 , or GLUT4 , have been described as candidates for cancer treatment. A better understanding of molecular recognition within this family is also useful for other applications, such as the design of GLUT1 and GLUT5 ligands as molecular probes for breast cancer diagnostics. − …”

A comprehensive overview of substrate specificity of glycoside hydrolases and transporters in the small intestine

“…13 Radiolabelled (a) and fluorescent probes (b) used in GLUT5 mediated uptake studies. References used: ref 1 [120], ref 2 [121], ref 3 [122], ref 4 [123], ref 5 [124], ref 6 [125], ref 7 [126], ref 8 [127], ref 9 [128] for rapid detection in breast cancer models [128]. A significant difference was found in the cancerous cells MCF-7 and MCF10AneoT compared to normal breast cells MCF10A underlining their potential for rapid on-site high-throughput diagnostics.…”