Pharmacologic Induction of Tolerance for Hypoxia & Hypothermia

Description

The investigator’s strategy is to: (a) repurpose approved drugs with potential for salutary effect upon RBC performance attributes that contribute to O2 delivery homeostasis during stress and (b) efficiently identify lead candidates through sequential evaluation in relevant and rigorous benchtop and in vivo models that include examination for gender-specific effects. Our assay platforms are selected to characterize RBC physiology relevant to O2 delivery, with focus upon RBC O2 affinity, energetics, biomechanics, vascular interaction and control of regional blood flow. The investigator will sample RBC suspensions serially (0, 1,3h) and quantify the RBC performance attributes across the range of modeled environmental extremes, defining RBC performance constraint as > 20% impairment in each attribute and determine pharmacologic rescue (defined as > 20% improvement in each attribute) using mixed model RM-ANOVA, as a function of gender. The investigator will power analysis to 80% at a<5%; based on our published data with these assays and experience, this requires 10-15 subjects/group. Drug candidates with evidence of PhIT-HyHo potential will advance to in vivo screening, prioritized by the number of RBC attributes rescued per drug. Specific description of our approach to evaluate each RBC performance attribute follows.

RBC performance attributes will be quantified under controlled conditions (Temperature: 28 – 37°C; pO2: 50 – 100 Torr, alone and in combination in our temperature-controlled thin film tonometer38 (NB temperature simulates hypothermic body temperature): (a) glycolytic flux, (b) resilience to oxidative stress, (c) deformability & aggregation, (d) O2 affinity and Bohr effect, (e) vasoactivity.