Accumulating evidence suggests oxidative stress is responsible for altered metabolic functions in various pathophysiological conditions including cancer, cardiovascular disease and aging. The production of reactive oxygen species (ROS) and subsequent oxidative damage suffered by the cells are accrued over time during normal aging have been implicated in the age related increase in many type of malignancies, including prostate cancer. Our preliminary experiments and current literature indicate activation of NF-kB, a redox-sensitive transcription factor, in presence of ROS and hence oxidant stress. Although the mechanism of action is still uncertain, we hypothesize that NF-kB, a redox-sensitive transcription factor, can serve as a useful biomarker for cellular oxidative stress, provided a rapid, sensitive and reliable assay for its measurement is available. To test this hypothesis, we plan to (a) develop a novel and ultra-sensitive ELISA to replace the tedious conventional electrophoretic mobility shift assay (EMSA) for evaluating the DNA binding activity of NF-kB, (b) establish whether the DNA binding activity of this transcription factor could be used as a marker for cellular oxidative stress and (c) validate and compare results using the NF-kB ELISA with established biomarkers for systemic oxidative stress in the serum of human volunteers. The proposed ELISA is expected to provide a rapid, sensitive micro-assay for the DNA binding activity of NF-kB that could be used practically in a clinical setting for evaluating cellular oxidative stress.
Oxidative stress is increasingly being recognized as a major factor in the development of human diseases, particularly diseases associated with aging such as arthritis, cardiovascular disease and cancer. The development of a facile and effective oxidative stress test would enable physicians to gauge the beneficial effects of dietary antioxidant supplements. It also would enable clinicians to monitor the therapeutic impact of antioxidant drugs currently under development and gain important feedback about the control of diseases associated with free radical damage. The potential market for these applications is substantial, with estimates ranging from $500 million to a billion or more.
| Gupta, Smiti V; McGowen, Richard M; Callewaert, Denis M et al. (2005) Quantitative chemiluminescent immunoassay for NF-kappaB-DNA binding activity. J Immunoassay Immunochem 26:125-43 |
