Redox Trapping for Biospecimen Preservation and Innovation in Sepsis Care

Wake Forest University Health Sciences, Winston-Salem NC

Investigators

Abstract

ABSTRACT (R21/R33 Grant) We aim to advance sepsis research and clinical diagnosis by introducing a new redox trapping formulation for preservation of blood specimens. The new formulation is anticipated: 1) to prevent artifactual oxidation and loss of specimen integrity during storage, 2) to enable studies of redox metabolism which underlines the dysregulated immunometabolic response in sepsis, and 3) to improve analysis of differentiating redox molecular markers associated with demographic features. Our combined expertise of redox biochemistry and innovation in patented redox chemical reagents, sepsis mechanisms, trial design and clinical expertise will be applied first to validate the new redox trapping formulation (R21 Phase) and then to scale-up by deploying this for collection of blood specimens at sites participating in the EMbedded Precision in Acute Care Trials (EMPACT) Network (R33 Phase). This is a newly formed network of premier critical care clinical trials sites designed to conduct precision clinical trials and discovery research in sepsis. Successful accomplishment of our aims will improve staging of septic patients and clinical precision research for discovery of new therapeutic targets for treatment of sepsis. More specifically, during the R21 Phase we will investigate the compatibility of the new redox formulation with high-end mass spectrometry and single cell technologies to evaluate its performance for measurement of redox and other biomarkers. The new formulation will be compared with current standard of care procedures for blood collection for clinical and research purposes (gold standard). To further validate this formula in sepsis patients, we will then evaluate its effects on standard clinical lab tests in a pilot study using prospectively collected paired single timepoint samples from critically ill patients with sepsis. We will further determine the advantage of redox trapping formulation by linking the measurements to physiologic and outcome data collected passively through the Wake Forest Critical Care Data Analytic Platform. Upon accomplishment of key metrics at the completion of R21 Phase, we will work with the EMPACT Network in the R33 Phase to collect blood specimens from 150 patients at 3 timepoints across the continuum of their disease progression using the standard methods and the new redox formulation. Analysis using high-end omics technologies will produce information-rich and redox-specific datasets, which will be integrated using bioinformatics approaches and linked to clinical data captured passively via the EMPACT data warehouse to generate new hypotheses for sepsis research. Lastly, we include a plan to share cryopreserved specimens, clinical and molecular data with investigators at the institution to support discovery or hypothesis-driven research through pilot mechanisms under the Center for Redox Biology and Medicine and Critical Illness, Injury and Research Recovery Research Center directed by MPIs Furdui and Files, respectively, and with outside investigators via EMPACT. ABSTRACT (R33 Phase) We aim to advance sepsis research and clinical diagnosis by introducing a new redox-based formulation for preservation of blood specimens. While blood is one of the top specimens for research and diagnostic use, it is highly unstable (metabolically active post-collection) and prone to artifactual oxidative damage due to the high iron content (Fenton chemistry) and metabolism. We envision our proposed formulation to advance clinical sepsis research and clinical care, to reduce pre-analytical variability in specimens’ analysis by preventing artifactual oxidation and loss of specimen integrity during storage, to augment discovery of new biomarkers for improved staging of patients, to improve analysis of differentiating redox molecular markers associated with demographic features, and to aid in development of new treatment regimens. During the R21 Phase, we applied our combined expertise of redox biochemistry and innovation in patented redox chemical reagents, sepsis mechanisms, trial design and clinical expertise to optimize the composition of the new redox formulation and to validate it in sepsis patients. Our efforts have been overwhelmingly successful as presented in the Progress Report. During the R33 Phase we will scale-up the studies by enrolling sepsis patients at minimum five hospital locations known for their ability to conduct premier critical care clinical trials and research studies in sepsis. The increased number of patients (total 225) will allow us to link the analytical measurements to physiologic and outcome data and thus firmly establish the value of redox formulation relative to standard collection procedures in quenching the endogenous molecular profile of specimens. Blood specimens will be collected from 150 patients at 3 timepoints along the continuum of their disease progression using the standard methods and the new redox formulation. Analysis using high-end omics technologies will produce information-rich and redox-specific datasets, which will be integrated using bioinformatics approaches and linked to clinical data captured passively at each location (e.g., at Wake using the Wake Forest Critical Care Data Analytic Platform) to generate new hypotheses for sepsis research. Lastly, we include a plan to share cryopreserved specimens, clinical and molecular data with investigators at other institutions to support discovery or hypothesis-driven research. Internally, this activity will be funded through pilot mechanisms under the Center for Redox Biology and Medicine and Critical Illness, Injury and Research Recovery Research Center directed by MPIs Furdui and Files, respectively.Â