The measurement of testosterone levels is central to the diagnosis of androgen disorders in men and women. Circulating testosterone is bound largely to sex hormone binding globulin (SHBG) and albumin;only the unbound or free fraction is biologically active. Therefore, in conditions that affect SHBG concentrations, such as aging, obesity, and diabetes, the determination of free testosterone is necessary to obtain an accurate assessment of androgen status. The current methods for the determination of free testosterone - equilibrium dialysis, ultrafiltration, tracer analog methods, and the use of algorithms based on the law-of-mass-action - suffer from problems of imprecision or inaccuracy. Equilibrium dialysis method, widely considered the reference method, is labor-intensive and cumbersome, and susceptible to errors due to tracer impurities, leading the Endocrine Society's Expert Panel to conclude that "...the calculation of free testosterone is the most useful estimate of free testosterone in plasma..." Therefore, algorithms for calculating free testosterone concentrations from total testosterone, SHBG and albumin concentrations using the law-of-mass-action equations have been used widely. Our preliminary data show that the current model of homogeneous testosterone binding sites on SHBG - the conceptual basis of the extant law-of-mass action equations - is erroneous, and that the free testosterone concentrations derived from the available law-of-mass action equations differ substantially from those measured by equilibrium dialysis.. Because of these limitations of the available methods, there is an enormous unmet need for an accurate method to determine free testosterone concentrations that can be applied conveniently in clinical practice. New evidence shows that testosterone's binding to SHBG conforms to an Ensemble Allostery Model (EAM). Based on this new EAM model of testosterone binding to SHBG, we have constructed a novel algorithm for calculating free testosterone levels that provides excellent conformity with values determined by equilibrium dialysis. This phase I SBIR application aims to characterize the technical performance of the novel algorithm by determining its precision, accuracy, functional sensitivity, intra- and inter-assay variation, and specificity, stability, and matrix effects (Aim 1).
Aim 2 will incorporate SHB genotype in the algorithm and apply it to men with different SHBG genotypes.
Aim 3 will characterize the performance of the assay in young and older men and women, and in obese men with alterations in SHBG concentrations. The product of the phase I studies will be a copyright-protected validated algorithm available as a stand-alone application that physicians can download on their personal computers or hand-held devices for use in their office or even on the bedside. An interdisciplinary team of investigators, strong preliminary data, and a validation plan that conforms to FDA guidance will assure the successful generation of a product for which there is considerable unmet need and which will advance our understanding of testosterone's binding to SHBG.

Public Health Relevance

The determination of free testosterone is necessary to obtain an accurate assessment of androgen status in the diagnosis and treatment of androgen disorders in men and women;however, because of the limitations of the available methods, there is an enormous unmet need for an accurate method to determine free testosterone concentrations that can be applied conveniently in clinical practice. Our preliminary data show that the current model of testosterone's binding to sex hormone binding globulin (SHBG) - the conceptual basis of the extant law-of-mass-action equations - is inaccurate;based on a new model of testosterone's binding to SHBG, we have constructed a novel algorithm for calculating free testosterone levels that provides excellent conformity with values determined by equilibrium dialysis. The product of this SBIR phase I application will be a copyright-protected validated novel algorithm for determination of free testosterone concentrations, which the physicians can download as a stand-alone application on their personal computers or hand-held devices for convenient use in their office or even on the bedside.

Agency
National Institute of Health (NIH)
Type
Small Business Innovation Research Grants (SBIR) - Phase I (R43)
Project #
1R43AG045011-01A1
Application #
8647313
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Joseph, Lyndon
Project Start
Project End
Budget Start
Budget End
Support Year
1
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Function Promoting Therapies, LLC
Department
Type
DUNS #
City
Boston
State
MA
Country
United States
Zip Code
02115