madman
Super Moderator
Dr. Jasuja and Dr. Bhasin's TruT project still active.
Much more to the story here.
Big things coming!
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Abstract Text
* Through detailed biophysical characterization, we have provided significant insights into the complex interactions among estradiol, testosterone, and SHBG by resolving the structural basis of SHBG allostery and hormone- binding interactions using steady-state equilibrium dialysis, Cryo-EM, stopped-flow kinetics, and computational modeling. This originally led to the deployment of TruE2TM algorithm for accurate determination of free testosterone concentrations. By incorporating E2-SHBG binding dynamics and the competitive interactions of testosterone, we have now developed a TruE2TM framework for accurate fE2 determination; this method is patent-protected
* Accordingly, Aim 1 will further characterize of the technical performance of the TruE2TM algorithm by determining its precision, accuracy, functional sensitivity, intra- and inter-assay variation over the range of physiologically relevant concentrations, specificity (the influence of other steroid hormones), stability, and matrix effects in diverse populations of men and women. The deliverable will be a comprehensive characterization of the algorithm in conformity with FDA guidelines. Aim 2 will focus on the performance of the TruE2TM method in comparison to the reference method (equilibrium dialysis) using serum isolated from two groups of men with gynecomastia with normal E2 such as a) men with hyperthyroidism who develop gynecomastia, b) men undergoing androgen deprivation therapy (ADT), who develop gynecomastia. A successful Phase I application will lead to the further optimization and validation of the TruE2TM method and facilitate its advancement to subsequent Phase II studies for real-world prospective validation and HIPAA-compliant implementation into clinical workflows in multiple hospitals in the US.
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PROJECT SUMMARY
Circulating estradiol (E2) plays an important role in human health and disease. In blood, E2 is bound with high affinity to sex hormone binding globulin (SHBG) and with substantially lower affinity to albumin; only the free fraction (fE2) is biologically active. Therefore, in many hormonal disorders characterized by alterations in fE2 and/ or binding protein concentrations, such as gynecomastia, pubertal disorders, hormone suppressing therapy of women with breast cancer, and in vitro fertilization, the determination of fE2 becomes important in the diagnoses, treatment, and monitoring. The current methods for the determination of fE2 have significant limitations. The tracer analog methods have been shown to be inaccurate, and the equilibrium dialysis method, the reference method, is difficult to perform and is not available to most clinicians. Therefore, algorithms for estimating fE2 have been advocated. However, we show that the prevailing algorithms based on linear models of E2 binding to SHBG with a fixedKd are erroneous and do not account for the allosteric coupling within the SHBG dimer nor consider the influence of circulating testosterone on E2's binding to SHBG. Through detailed biophysical characterization, we have provided significant insights into the complex interactions among estradiol, testosterone, and SHBG by resolving the structural basis of SHBG allostery and hormone- binding interactions using steady-state equilibrium dialysis, Cryo-EM, stopped-flow kinetics, and computational modeling. This originally led to the deployment of TruE2TM algorithm for accurate determination of free testosterone concentrations. By incorporating E2-SHBG binding dynamics and the competitive interactions of testosterone, we have now developed a TruE2TM framework for accurate fE2 determination; this method is patent-protected. The current Phase I SBIR grant aims to further optimize and validate its performance in healthy men and women and in clinical conditions with alterations in fE2 levels. Accordingly, Aim 1 will further characterize of the technical performance of the TruE2TM algorithm by determining its precision, accuracy, functional sensitivity, intra- and inter-assay variation over the range of physiologically relevant concentrations, specificity (the influence of other steroid hormones), stability, and matrix effects in diverse populations of men and women. The deliverable will be a comprehensive characterization of the algorithm in conformity with FDA guidelines. Aim 2 will focus on the performance of the TruE2TM method in comparison to the reference method (equilibrium dialysis) using serum isolated from two groups of men with gynecomastia with normal E2 such as a) men with hyperthyroidism who develop gynecomastia, b) men undergoing androgen deprivation therapy (ADT), who develop gynecomastia. A successful Phase I application will lead to the further optimization and validation of the TruE2TM method and facilitate its advancement to subsequent Phase II studies for real-world prospective validation and HIPAA-compliant implementation into clinical workflows in multiple hospitals in the US.
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PROJECT NARRATIVE
Estradiol plays an important role in health and disease across the human lifespan. Because E2 circulates in blood, largely bound to proteins, the accurate measurement of its biologically active fraction free estradiol is central to the rational diagnoses and management of reproductive and hormonal disorders in men and women; however, methods for accurate determination of free estradiol concentrations are not readily available to clinicians. By integrating insights from the dynamic and allosteric binding of estradiol to sex hormone-binding globulin (SHBG), XYone has developed a novel TruE2TM method for free estradiol determination. The current Phase I grant aims to further optimize and validate the TruE2TM method for the determination of free estradiol concentrations in men and women with clinical conditions characterized by altered binding protein and free estradiol concentrations.
Much more to the story here.
Big things coming!
=============
Abstract Text
* Through detailed biophysical characterization, we have provided significant insights into the complex interactions among estradiol, testosterone, and SHBG by resolving the structural basis of SHBG allostery and hormone- binding interactions using steady-state equilibrium dialysis, Cryo-EM, stopped-flow kinetics, and computational modeling. This originally led to the deployment of TruE2TM algorithm for accurate determination of free testosterone concentrations. By incorporating E2-SHBG binding dynamics and the competitive interactions of testosterone, we have now developed a TruE2TM framework for accurate fE2 determination; this method is patent-protected
* Accordingly, Aim 1 will further characterize of the technical performance of the TruE2TM algorithm by determining its precision, accuracy, functional sensitivity, intra- and inter-assay variation over the range of physiologically relevant concentrations, specificity (the influence of other steroid hormones), stability, and matrix effects in diverse populations of men and women. The deliverable will be a comprehensive characterization of the algorithm in conformity with FDA guidelines. Aim 2 will focus on the performance of the TruE2TM method in comparison to the reference method (equilibrium dialysis) using serum isolated from two groups of men with gynecomastia with normal E2 such as a) men with hyperthyroidism who develop gynecomastia, b) men undergoing androgen deprivation therapy (ADT), who develop gynecomastia. A successful Phase I application will lead to the further optimization and validation of the TruE2TM method and facilitate its advancement to subsequent Phase II studies for real-world prospective validation and HIPAA-compliant implementation into clinical workflows in multiple hospitals in the US.
==
PROJECT SUMMARY
Circulating estradiol (E2) plays an important role in human health and disease. In blood, E2 is bound with high affinity to sex hormone binding globulin (SHBG) and with substantially lower affinity to albumin; only the free fraction (fE2) is biologically active. Therefore, in many hormonal disorders characterized by alterations in fE2 and/ or binding protein concentrations, such as gynecomastia, pubertal disorders, hormone suppressing therapy of women with breast cancer, and in vitro fertilization, the determination of fE2 becomes important in the diagnoses, treatment, and monitoring. The current methods for the determination of fE2 have significant limitations. The tracer analog methods have been shown to be inaccurate, and the equilibrium dialysis method, the reference method, is difficult to perform and is not available to most clinicians. Therefore, algorithms for estimating fE2 have been advocated. However, we show that the prevailing algorithms based on linear models of E2 binding to SHBG with a fixedKd are erroneous and do not account for the allosteric coupling within the SHBG dimer nor consider the influence of circulating testosterone on E2's binding to SHBG. Through detailed biophysical characterization, we have provided significant insights into the complex interactions among estradiol, testosterone, and SHBG by resolving the structural basis of SHBG allostery and hormone- binding interactions using steady-state equilibrium dialysis, Cryo-EM, stopped-flow kinetics, and computational modeling. This originally led to the deployment of TruE2TM algorithm for accurate determination of free testosterone concentrations. By incorporating E2-SHBG binding dynamics and the competitive interactions of testosterone, we have now developed a TruE2TM framework for accurate fE2 determination; this method is patent-protected. The current Phase I SBIR grant aims to further optimize and validate its performance in healthy men and women and in clinical conditions with alterations in fE2 levels. Accordingly, Aim 1 will further characterize of the technical performance of the TruE2TM algorithm by determining its precision, accuracy, functional sensitivity, intra- and inter-assay variation over the range of physiologically relevant concentrations, specificity (the influence of other steroid hormones), stability, and matrix effects in diverse populations of men and women. The deliverable will be a comprehensive characterization of the algorithm in conformity with FDA guidelines. Aim 2 will focus on the performance of the TruE2TM method in comparison to the reference method (equilibrium dialysis) using serum isolated from two groups of men with gynecomastia with normal E2 such as a) men with hyperthyroidism who develop gynecomastia, b) men undergoing androgen deprivation therapy (ADT), who develop gynecomastia. A successful Phase I application will lead to the further optimization and validation of the TruE2TM method and facilitate its advancement to subsequent Phase II studies for real-world prospective validation and HIPAA-compliant implementation into clinical workflows in multiple hospitals in the US.
====
PROJECT NARRATIVE
Estradiol plays an important role in health and disease across the human lifespan. Because E2 circulates in blood, largely bound to proteins, the accurate measurement of its biologically active fraction free estradiol is central to the rational diagnoses and management of reproductive and hormonal disorders in men and women; however, methods for accurate determination of free estradiol concentrations are not readily available to clinicians. By integrating insights from the dynamic and allosteric binding of estradiol to sex hormone-binding globulin (SHBG), XYone has developed a novel TruE2TM method for free estradiol determination. The current Phase I grant aims to further optimize and validate the TruE2TM method for the determination of free estradiol concentrations in men and women with clinical conditions characterized by altered binding protein and free estradiol concentrations.
