The RAAS is targeted in therapies that reduce mortality. However, increasing evidence suggests that improvement is independent of reductions in blood pressure or changes in angiotensin II. In fact, a strong link between an activated RAAS and cytokine and other inflammation markers is becoming more apparent as an initiating factor in early hypertension, atherosclerosis and diabetes. Thus, better understanding of the entire RAAS as well as a more extensive complement of other hormonal systems interacting to control blood pressure and influence target organ damage is needed. At present, full assessment of the hormonal systems involved is difficult and costly, in terms of volume of sample and expense of the multiple assays required. Thus, our desire to understand complex interactions among RAAS and associated systems in cardiovascular (CV) pathology has driven us to consider new technologies. The goal of this proposal is to develop new tools for unified assessment of these important hormonal systems, based on the premise that we adapt conventional ELISAs and radioimmunoassays for use with a new technology, the Luminex LabMAP system. This will reduce sample volumes required for multiple individual assays and ultimately reduce complexity and cost of assessing multiple systems. In addition to quantitative assays, we will develop semi-quantitative protein suspension microarrays using the same particle based technology to expedite verification of gene arrays and other molecular techniques that document patterns of activation or inhibition of mechanisms involved in CV disease. We anticipate that application of the new technology will simplify assessment of RAAS and other associated hormonal systems activated synchronously or in opposition. In addition, reduced sample volumes and costs should also allow expansion of both clinical and basic science studies to include more realistic assessment of major hormonal systems involved in CV disease. Finally, an advantage is that it requires relatively inexpensive instrumentation, and provides greater flexibility, quantification, and sensitivity compared with fixed platform protein arrays currently available. As such the peptide and protein applications developed in this proposal will be ideal for widespread use.

R21RR018370

2003-08-04

2006-07-31