Chronic diseases are the leading cause of death in the world, accounting for over 60 % of all fatalities. Decreased levels of the tripeptide glutathione (GSH) are well-known to be associated with chronic diseases and are of interest in monitoring disease progression and therapeutic interventions. For example, recent studies show that monitoring whole blood glutathione (GSH) levels can be of benefit towards diagnosing asymptomatic heart failure and mitochondrial disorders. GSH monitoring is also useful in the health assessment of patients with chronic renal disease or post-operative open heart surgery. However, reported levels of GSH in both healthy subjects and patients exhibit inter-laboratory variation. Plasma GSH levels are often reported despite the fact that they account for less than 0.5 % of the circulating GSH. Divergent sample processing and conjugation steps propagate inter-laboratory variability, as they result in varying degrees of sample oxidation and labeling. In order to achieve consistent and efficient GSH monitoring, the major goal of this proposal is to develop a fluorescent indicator that functions directly in minimally-diluted whole blood. In order to achieve this goal, the following specific aims are proposed: 1. Design, synthesis and evaluation of NIR-active GSH-selective indicators. Based on the preliminary data referred to above, our working hypothesis is that highly selective indicators for GSH can be designed to bind GSH via multipoint supramolecular interactions and produce concomitant fluorescence signals without optical interference from hemoglobin or other blood components. 2. Develop methodology for detection and quantitation of GSH in whole blood. We postulate, again on the basis of our preliminary data, that selectivity and sensitivity for GSH over a disease-relevant range can be achieved in human blood via the design of GSH-selective NIR active probe-containing receptors. The studies proposed in this aim are thus focused on optimization and demonstration of the utility of the proposed design principles and synthetic indicators via extensive evaluation and validation studies.

Public Health Relevance

Glutathione is an amino acid that plays a significant role in human health. It is used to monitor the progression and treatment of chronic diseases, the cause of over 60 % of all deaths worldwide. The goal of this proposal is to develop a chemical indicator that changes its optical properties in response to glutathione levels without using any separations or expensive and fragile biological components. This would be directly analogous to using a pH indicator, but instead of detecting protons we plan to detect an amino acid. Diagnostic tests for glutathione do already exist. However, the testing is relatively expensive. Due to the importance of glutathione, other researchers have also worked on developing simple and inexpensive indicator methods;however, these at very best are potentially useful in plasma measurements and currently not whole blood. The measurement of plasma glutathione levels is indeed useful in many cases but is often not correlated to the same specific disease states as are whole blood glutathione levels. .

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Academic Research Enhancement Awards (AREA) (R15)
Project #
1R15EB016870-01A1
Application #
8627059
Study Section
Synthetic and Biological Chemistry A Study Section (SBCA)
Program Officer
Erim, Zeynep
Project Start
2014-04-01
Project End
2017-03-31
Budget Start
2014-04-01
Budget End
2017-03-31
Support Year
1
Fiscal Year
2014
Total Cost
$434,644
Indirect Cost
$118,746
Name
Portland State University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
052226800
City
Portland
State
OR
Country
United States
Zip Code
97207
Barve, Aabha; Lowry, Mark; Escobedo, Jorge O et al. (2014) Differences in heterocycle basicity distinguish homocysteine from cysteine using aldehyde-bearing fluorophores. Chem Commun (Camb) 50:8219-22