title: A cell-based phenotypic assay to identify cardioprotective agents.
abstract: BACKGROUND
  Tissue ischemia/reperfusion (IR) injury underlies several leading causes of death such as heart-attack and stroke. The lack of clinical therapies for IR injury may be partly due to the difficulty of adapting IR injury models to high-throughput screening (HTS).
  
  
  OBJECTIVE
  To develop a model of IR injury that is both physiologically relevant and amenable to HTS.
  
  
  RESULTS
  A microplate-based respirometry apparatus was used. Controlling gas flow in the plate head space, coupled with the instrument's mechanical systems, yielded a 24-well model of IR injury in which H9c2 cardiomyocytes were transiently trapped in a small volume, rendering them ischemic. After initial validation with known protective molecules, the model was used to screen a 2000-molecule library, with post-IR cell death as an end point. Po2 and pH monitoring in each well also afforded metabolic data. Ten protective, detrimental, and inert molecules from the screen were subsequently tested in a Langendorff-perfused heart model of IR injury, revealing strong correlations between the screening end point and both recovery of cardiac function (negative, r2=0.66) and infarct size (positive, r2=0.62). Relationships between the effects of added molecules on cellular bioenergetics and protection against IR injury were also studied.
  
  
  CONCLUSIONS
  This novel cell-based assay can predict either protective or detrimental effects on IR injury in the intact heart. Its application may help identify therapeutic or harmful molecules.
authors.affiliation: School of Medicine, University of Rochester Medical Center, 601 Elmwood Ave, Rochester, NY 14642, USA.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Guo
authors.forename: Adam
authors.initials: A
authors.lastname: Olm-Shipman
authors.forename: Andrew
authors.initials: A
authors.lastname: Walters
authors.forename: William R
authors.initials: WR
authors.lastname: Urciuoli
authors.forename: Stefanie
authors.initials: S
authors.lastname: Devito
authors.forename: Sergiy M
authors.initials: SM
authors.lastname: Nadtochiy
authors.forename: Andrew P
authors.initials: AP
authors.lastname: Wojtovich
authors.forename: Paul S
authors.initials: PS
authors.lastname: Brookes
mesh_terms: D000818:Animals; D002316:Cardiotonic Agents / Q000494:pharmacology / Q000627:therapeutic use*; D016923:Cell Death; D002460:Cell Line; D002478:Cells, Cultured; D004195:Disease Models, Animal; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D015428:Myocardial Reperfusion Injury / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D010641:Phenotype; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results
chemical_list: D002316:Cardiotonic Agents