title: Sucrose Nonfermenting-Related Kinase Enzyme-Mediated Rho-Associated Kinase Signaling is Responsible for Cardiac Function.
abstract: BACKGROUND
  Cardiac metabolism is critical for the functioning of the heart, and disturbance in this homeostasis is likely to influence cardiac disorders or cardiomyopathy. Our laboratory has previously shown that SNRK (sucrose nonfermenting related kinase) enzyme, which belongs to the AMPK (adenosine monophosphate-activated kinase) family, was essential for cardiac metabolism in mammals. Snrk global homozygous knockout (KO) mice die at postnatal day 0, and conditional deletion of Snrk in cardiomyocytes (Snrk cmcKO) leads to cardiac failure and death by 8 to 10 months.
  
  
  RESULTS
  We performed additional cardiac functional studies using echocardiography and identified further cardiac functional deficits in Snrk cmcKO mice. Nuclear magnetic resonance-based metabolomics analysis identified key metabolic pathway deficits in SNRK knockdown cardiomyocytes in vitro. Specifically, metabolites involved in lipid metabolism and oxidative phosphorylation are altered, and perturbations in these pathways can result in cardiac function deficits and heart failure. A phosphopeptide-based proteomic screen identified ROCK (Rho-associated kinase) as a putative substrate for SNRK, and mass spec-based fragment analysis confirmed key amino acid residues on ROCK that are phosphorylated by SNRK. Western blot analysis on heart lysates from Snrk cmcKO adult mice and SNRK knockdown cardiomyocytes showed increased ROCK activity. In addition, in vivo inhibition of ROCK partially rescued the in vivo Snrk cmcKO cardiac function deficits.
  
  
  CONCLUSIONS
  Collectively, our data suggest that SNRK in cardiomyocytes is responsible for maintaining cardiac metabolic homeostasis, which is mediated in part by ROCK, and alteration of this homeostasis influences cardiac function in the adult heart.
authors: From the Department of Pediatrics (S.M.C., A.G., S. Pooya, M.B., R.R.), OBGYN, Developmental Vascular Biology Program, Children's Research Institute (R.R.), Division of Cardiovascular Medicine, Cardiovascular Center, Clinical and Translational Science Institute (L.M.H.), Division of Cardiovascular Medicine, Department of Cell Biology, Neurobiology and Anatomy, Cardiovascular Center, Clinical and Translational Science Institute (J.L.S.), and Division of Pediatric Pathology, Department of Pathology (S.N.K.), Medical College of Wisconsin, Milwaukee; Department of Chemical and Biological Engineering (V.J.B., X.B., S. Palecek), Morgridge Institute for Research (M.A.H.), University of Wisconsin-Madison; Marginalen Bank, Stockholm, Sweden (I.S.); Department of Chemistry and Biochemistry, University of Wisconsin-Milwaukee (S.P.M.).
authors: Stephanie M
authors: SM
authors: Cossette
authors: From the Department of Pediatrics (S.M.C., A.G., S. Pooya, M.B., R.R.), OBGYN, Developmental Vascular Biology Program, Children's Research Institute (R.R.), Division of Cardiovascular Medicine, Cardiovascular Center, Clinical and Translational Science Institute (L.M.H.), Division of Cardiovascular Medicine, Department of Cell Biology, Neurobiology and Anatomy, Cardiovascular Center, Clinical and Translational Science Institute (J.L.S.), and Division of Pediatric Pathology, Department of Pathology (S.N.K.), Medical College of Wisconsin, Milwaukee; Department of Chemical and Biological Engineering (V.J.B., X.B., S. Palecek), Morgridge Institute for Research (M.A.H.), University of Wisconsin-Madison; Marginalen Bank, Stockholm, Sweden (I.S.); Department of Chemistry and Biochemistry, University of Wisconsin-Milwaukee (S.P.M.).
authors: Vijesh J
authors: VJ
authors: Bhute
authors: From the Department of Pediatrics (S.M.C., A.G., S. Pooya, M.B., R.R.), OBGYN, Developmental Vascular Biology Program, Children's Research Institute (R.R.), Division of Cardiovascular Medicine, Cardiovascular Center, Clinical and Translational Science Institute (L.M.H.), Division of Cardiovascular Medicine, Department of Cell Biology, Neurobiology and Anatomy, Cardiovascular Center, Clinical and Translational Science Institute (J.L.S.), and Division of Pediatric Pathology, Department of Pathology (S.N.K.), Medical College of Wisconsin, Milwaukee; Department of Chemical and Biological Engineering (V.J.B., X.B., S. Palecek), Morgridge Institute for Research (M.A.H.), University of Wisconsin-Madison; Marginalen Bank, Stockholm, Sweden (I.S.); Department of Chemistry and Biochemistry, University of Wisconsin-Milwaukee (S.P.M.).
authors: Xiaoping
authors: X
authors: Bao
authors: From the Department of Pediatrics (S.M.C., A.G., S. Pooya, M.B., R.R.), OBGYN, Developmental Vascular Biology Program, Children's Research Institute (R.R.), Division of Cardiovascular Medicine, Cardiovascular Center, Clinical and Translational Science Institute (L.M.H.), Division of Cardiovascular Medicine, Department of Cell Biology, Neurobiology and Anatomy, Cardiovascular Center, Clinical and Translational Science Institute (J.L.S.), and Division of Pediatric Pathology, Department of Pathology (S.N.K.), Medical College of Wisconsin, Milwaukee; Department of Chemical and Biological Engineering (V.J.B., X.B., S. Palecek), Morgridge Institute for Research (M.A.H.), University of Wisconsin-Madison; Marginalen Bank, Stockholm, Sweden (I.S.); Department of Chemistry and Biochemistry, University of Wisconsin-Milwaukee (S.P.M.).
authors: Leanne M
authors: LM
authors: Harmann
authors: From the Department of Pediatrics (S.M.C., A.G., S. Pooya, M.B., R.R.), OBGYN, Developmental Vascular Biology Program, Children's Research Institute (R.R.), Division of Cardiovascular Medicine, Cardiovascular Center, Clinical and Translational Science Institute (L.M.H.), Division of Cardiovascular Medicine, Department of Cell Biology, Neurobiology and Anatomy, Cardiovascular Center, Clinical and Translational Science Institute (J.L.S.), and Division of Pediatric Pathology, Department of Pathology (S.N.K.), Medical College of Wisconsin, Milwaukee; Department of Chemical and Biological Engineering (V.J.B., X.B., S. Palecek), Morgridge Institute for Research (M.A.H.), University of Wisconsin-Madison; Marginalen Bank, Stockholm, Sweden (I.S.); Department of Chemistry and Biochemistry, University of Wisconsin-Milwaukee (S.P.M.).
authors: Mark A
authors: MA
authors: Horswill
authors: From the Department of Pediatrics (S.M.C., A.G., S. Pooya, M.B., R.R.), OBGYN, Developmental Vascular Biology Program, Children's Research Institute (R.R.), Division of Cardiovascular Medicine, Cardiovascular Center, Clinical and Translational Science Institute (L.M.H.), Division of Cardiovascular Medicine, Department of Cell Biology, Neurobiology and Anatomy, Cardiovascular Center, Clinical and Translational Science Institute (J.L.S.), and Division of Pediatric Pathology, Department of Pathology (S.N.K.), Medical College of Wisconsin, Milwaukee; Department of Chemical and Biological Engineering (V.J.B., X.B., S. Palecek), Morgridge Institute for Research (M.A.H.), University of Wisconsin-Madison; Marginalen Bank, Stockholm, Sweden (I.S.); Department of Chemistry and Biochemistry, University of Wisconsin-Milwaukee (S.P.M.).
authors: Indranil
authors: I
authors: Sinha
authors: From the Department of Pediatrics (S.M.C., A.G., S. Pooya, M.B., R.R.), OBGYN, Developmental Vascular Biology Program, Children's Research Institute (R.R.), Division of Cardiovascular Medicine, Cardiovascular Center, Clinical and Translational Science Institute (L.M.H.), Division of Cardiovascular Medicine, Department of Cell Biology, Neurobiology and Anatomy, Cardiovascular Center, Clinical and Translational Science Institute (J.L.S.), and Division of Pediatric Pathology, Department of Pathology (S.N.K.), Medical College of Wisconsin, Milwaukee; Department of Chemical and Biological Engineering (V.J.B., X.B., S. Palecek), Morgridge Institute for Research (M.A.H.), University of Wisconsin-Madison; Marginalen Bank, Stockholm, Sweden (I.S.); Department of Chemistry and Biochemistry, University of Wisconsin-Milwaukee (S.P.M.).
authors: Adam
authors: A
authors: Gastonguay
authors: From the Department of Pediatrics (S.M.C., A.G., S. Pooya, M.B., R.R.), OBGYN, Developmental Vascular Biology Program, Children's Research Institute (R.R.), Division of Cardiovascular Medicine, Cardiovascular Center, Clinical and Translational Science Institute (L.M.H.), Division of Cardiovascular Medicine, Department of Cell Biology, Neurobiology and Anatomy, Cardiovascular Center, Clinical and Translational Science Institute (J.L.S.), and Division of Pediatric Pathology, Department of Pathology (S.N.K.), Medical College of Wisconsin, Milwaukee; Department of Chemical and Biological Engineering (V.J.B., X.B., S. Palecek), Morgridge Institute for Research (M.A.H.), University of Wisconsin-Madison; Marginalen Bank, Stockholm, Sweden (I.S.); Department of Chemistry and Biochemistry, University of Wisconsin-Milwaukee (S.P.M.).
authors: Shabnam
authors: S
authors: Pooya
authors: From the Department of Pediatrics (S.M.C., A.G., S. Pooya, M.B., R.R.), OBGYN, Developmental Vascular Biology Program, Children's Research Institute (R.R.), Division of Cardiovascular Medicine, Cardiovascular Center, Clinical and Translational Science Institute (L.M.H.), Division of Cardiovascular Medicine, Department of Cell Biology, Neurobiology and Anatomy, Cardiovascular Center, Clinical and Translational Science Institute (J.L.S.), and Division of Pediatric Pathology, Department of Pathology (S.N.K.), Medical College of Wisconsin, Milwaukee; Department of Chemical and Biological Engineering (V.J.B., X.B., S. Palecek), Morgridge Institute for Research (M.A.H.), University of Wisconsin-Madison; Marginalen Bank, Stockholm, Sweden (I.S.); Department of Chemistry and Biochemistry, University of Wisconsin-Milwaukee (S.P.M.).
authors: Michelle
authors: M
authors: Bordas
authors: From the Department of Pediatrics (S.M.C., A.G., S. Pooya, M.B., R.R.), OBGYN, Developmental Vascular Biology Program, Children's Research Institute (R.R.), Division of Cardiovascular Medicine, Cardiovascular Center, Clinical and Translational Science Institute (L.M.H.), Division of Cardiovascular Medicine, Department of Cell Biology, Neurobiology and Anatomy, Cardiovascular Center, Clinical and Translational Science Institute (J.L.S.), and Division of Pediatric Pathology, Department of Pathology (S.N.K.), Medical College of Wisconsin, Milwaukee; Department of Chemical and Biological Engineering (V.J.B., X.B., S. Palecek), Morgridge Institute for Research (M.A.H.), University of Wisconsin-Madison; Marginalen Bank, Stockholm, Sweden (I.S.); Department of Chemistry and Biochemistry, University of Wisconsin-Milwaukee (S.P.M.).
authors: Suresh N
authors: SN
authors: Kumar
authors: From the Department of Pediatrics (S.M.C., A.G., S. Pooya, M.B., R.R.), OBGYN, Developmental Vascular Biology Program, Children's Research Institute (R.R.), Division of Cardiovascular Medicine, Cardiovascular Center, Clinical and Translational Science Institute (L.M.H.), Division of Cardiovascular Medicine, Department of Cell Biology, Neurobiology and Anatomy, Cardiovascular Center, Clinical and Translational Science Institute (J.L.S.), and Division of Pediatric Pathology, Department of Pathology (S.N.K.), Medical College of Wisconsin, Milwaukee; Department of Chemical and Biological Engineering (V.J.B., X.B., S. Palecek), Morgridge Institute for Research (M.A.H.), University of Wisconsin-Madison; Marginalen Bank, Stockholm, Sweden (I.S.); Department of Chemistry and Biochemistry, University of Wisconsin-Milwaukee (S.P.M.).
authors: Shama P
authors: SP
authors: Mirza
authors: From the Department of Pediatrics (S.M.C., A.G., S. Pooya, M.B., R.R.), OBGYN, Developmental Vascular Biology Program, Children's Research Institute (R.R.), Division of Cardiovascular Medicine, Cardiovascular Center, Clinical and Translational Science Institute (L.M.H.), Division of Cardiovascular Medicine, Department of Cell Biology, Neurobiology and Anatomy, Cardiovascular Center, Clinical and Translational Science Institute (J.L.S.), and Division of Pediatric Pathology, Department of Pathology (S.N.K.), Medical College of Wisconsin, Milwaukee; Department of Chemical and Biological Engineering (V.J.B., X.B., S. Palecek), Morgridge Institute for Research (M.A.H.), University of Wisconsin-Madison; Marginalen Bank, Stockholm, Sweden (I.S.); Department of Chemistry and Biochemistry, University of Wisconsin-Milwaukee (S.P.M.).
authors: Sean P
authors: SP
authors: Palecek
authors: From the Department of Pediatrics (S.M.C., A.G., S. Pooya, M.B., R.R.), OBGYN, Developmental Vascular Biology Program, Children's Research Institute (R.R.), Division of Cardiovascular Medicine, Cardiovascular Center, Clinical and Translational Science Institute (L.M.H.), Division of Cardiovascular Medicine, Department of Cell Biology, Neurobiology and Anatomy, Cardiovascular Center, Clinical and Translational Science Institute (J.L.S.), and Division of Pediatric Pathology, Department of Pathology (S.N.K.), Medical College of Wisconsin, Milwaukee; Department of Chemical and Biological Engineering (V.J.B., X.B., S. Palecek), Morgridge Institute for Research (M.A.H.), University of Wisconsin-Madison; Marginalen Bank, Stockholm, Sweden (I.S.); Department of Chemistry and Biochemistry, University of Wisconsin-Milwaukee (S.P.M.).
authors: Jennifer L
authors: JL
authors: Strande
authors: From the Department of Pediatrics (S.M.C., A.G., S. Pooya, M.B., R.R.), OBGYN, Developmental Vascular Biology Program, Children's Research Institute (R.R.), Division of Cardiovascular Medicine, Cardiovascular Center, Clinical and Translational Science Institute (L.M.H.), Division of Cardiovascular Medicine, Department of Cell Biology, Neurobiology and Anatomy, Cardiovascular Center, Clinical and Translational Science Institute (J.L.S.), and Division of Pediatric Pathology, Department of Pathology (S.N.K.), Medical College of Wisconsin, Milwaukee; Department of Chemical and Biological Engineering (V.J.B., X.B., S. Palecek), Morgridge Institute for Research (M.A.H.), University of Wisconsin-Madison; Marginalen Bank, Stockholm, Sweden (I.S.); Department of Chemistry and Biochemistry, University of Wisconsin-Milwaukee (S.P.M.). rramchan@mcw.edu.
authors: Ramani
authors: R
authors: Ramchandran
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004452:Echocardiography; D053595:Embryonic Stem Cells / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology; D004734:Energy Metabolism / Q000187:drug effects; D005355:Fibrosis; D020022:Genetic Predisposition to Disease; D006333:Heart Failure / Q000201:enzymology* / Q000235:genetics / Q000473:pathology / Q000503:physiopathology; D006720:Homozygote; D061307:Human Umbilical Vein Endothelial Cells / Q000201:enzymology; D050356:Lipid Metabolism; D009682:Magnetic Resonance Spectroscopy; D055432:Metabolomics / Q000379:methods; D018345:Mice, Knockout; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology; D010085:Oxidative Phosphorylation; D010641:Phenotype; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D015398:Signal Transduction; D014162:Transfection; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D047428:Protein Kinase Inhibitors; C490993:Snrk protein, mouse; C490994:SNRK protein, human; D017346:Protein Serine-Threonine Kinases; C515851:ROCK1 protein, human; C515854:Rock1 protein, mouse; D054460:rho-Associated Kinases
keywords: MYH6; ROCK; SNRK; cardiac; echocardiography; metabolism