title: Marked hyperglycemia attenuates anesthetic preconditioning in human-induced pluripotent stem cell-derived cardiomyocytes.
abstract: BACKGROUND
  Anesthetic preconditioning protects cardiomyocytes from oxidative stress-induced injury, but it is ineffective in patients with diabetes mellitus. To address the role of hyperglycemia in the inability of diabetic individuals to be preconditioned, we used human cardiomyocytes differentiated from induced pluripotent stem cells generated from patients with or without type 2 diabetes mellitus (DM-iPSC- and N-iPSC-CMs, respectively) to investigate the efficacy of preconditioning in varying glucose conditions (5, 11, and 25 mM).
  
  
  METHODS
  Induced pluripotent stem cells were induced to generate cardiomyocytes by directed differentiation. For subsequent studies, cardiomyocytes were identified by genetic labeling with enhanced green fluorescent protein driven by a cardiac-specific promoter. Cell viability was analyzed by lactate dehydrogenase assay. Confocal microscopy was utilized to measure opening of the mitochondrial permeability transition pore and the mitochondrial adenosine 5'-triphosphate-sensitive potassium channels.
  
  
  RESULTS
  Isoflurane (0.5 mM) preconditioning protected N-iPSC- and DM-iPSC-CMs from oxidative stress-induced lactate dehydrogenase release and mitochondrial permeability transition pore opening in 5 mM and 11 mM glucose. Isoflurane triggered mitochondrial adenosine-5'-triphosphate-sensitive potassium channel opening in N-iPSC-CMs in 5 mM and 11 mM glucose and in DM-iPSC-CMs in 5 mM glucose; 25 mM glucose disrupted anesthetic preconditioning-mediated protection in DM-iPSC- and N-iPSC-CMs.
  
  
  CONCLUSIONS
  The opening of mitochondrial adenosine 5'-triphosphate-sensitive potassium channels are disrupted in DM-iPSC-CMs in 11 mM and 25 mM glucose and in N-iPSC-CMs in 25 mM glucose. Cardiomyocytes derived from healthy donors and patients with a specific disease, such as diabetes in this study, open possibilities in studying genotype- and phenotype-related pathologies in a human-relevant model.
authors: Department of Anesthesiology, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA. scanfiel@mcw.edu
authors: Scott G
authors: SG
authors: Canfield
authors: Ana
authors: A
authors: Sepac
authors: Filip
authors: F
authors: Sedlic
authors: Maria Y
authors: MY
authors: Muravyeva
authors: Xiaowen
authors: X
authors: Bai
authors: Zeljko J
authors: ZJ
authors: Bosnjak
mesh_terms: D000777:Anesthetics / Q000494:pharmacology*; D018685:Anesthetics, Inhalation / Q000494:pharmacology; D024722:Cardiac Myosins / Q000235:genetics; D002454:Cell Differentiation / Q000187:drug effects; D003924:Diabetes Mellitus, Type 2 / Q000097:blood; D005347:Fibroblasts; D005455:Fluorescent Antibody Technique; D005822:Genetic Vectors; D006801:Humans; D006943:Hyperglycemia / Q000378:metabolism*; D007530:Isoflurane / Q000494:pharmacology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics; D008564:Membrane Potentials / Q000187:drug effects; D042282:Microdissection; D018613:Microscopy, Confocal; D051336:Mitochondrial Membranes / Q000187:drug effects; D032383:Myocytes, Cardiac / Q000187:drug effects*; D018994:Myosin Light Chains / Q000235:genetics; D010539:Permeability; D039904:Pluripotent Stem Cells / Q000187:drug effects*
chemical_list: D000777:Anesthetics; D018685:Anesthetics, Inhalation; D018994:Myosin Light Chains; C107184:myosin light chain 2; D007530:Isoflurane; D007770:L-Lactate Dehydrogenase; D024722:Cardiac Myosins