Thorac Cardiovasc Surg 2006; 54(6): 381-387
DOI: 10.1055/s-2006-924267
Basic Science

© Georg Thieme Verlag KG Stuttgart · New York

Differential Gene Expression in Response to Ventricular Unloading in Rat and Human Myocardium

T. Doenst1 , H. Bugger1 , S. Leippert1 , B. Barleon2 , D. Marme2 , F. Beyersdorf1
  • 1Department of Cardiovascular Surgery, Albert-Ludwigs University of Freiburg, Freiburg i. Br., Germany
  • 2Clinic for Tumor Biology, Albert-Ludwigs University of Freiburg, Freiburg i. Br., Germany
Further Information

Publication History

Received October 23, 2005

Publication Date:
07 September 2006 (online)

Abstract

Left ventricular unloading by mechanical assist devices induces myocardial atrophy. We aimed to systematically identify differentially expressed genes in a model of physiological atrophy (unloading of healthy rat myocardium) and compare these changes to those in a unloaded, failing human heart. Methods: Atrophy in rat hearts was induced by heterotopic transplantation of a donor heart into the abdomen of an isogenic recipient. After one week, donor and recipient RNA was isolated. Differential gene expression was assessed by subtractive hybridization. Two screens with radioactive probes were performed to verify differentially expressed clones. Positive clones were sequenced and cDNA of genes of known homology were used as probes for hybridization with RNA from separate atrophied rat hearts and human tissue from a normal, failing or failing and unloaded left ventricle. Results: We picked 1880 clones from the subtractive hybridization procedure (940/940: forward/reverse runs assessing up- or down-regulation, respectively). The first screen verified 465/140 and the second screen verified 67/30 clones. 24/23 clones were sequenced and 14/10 homologies to known genes were found. In the atrophied heart, respiratory chain and metabolic genes were down-regulated (NADH‐DH, cytochrome c oxidase, acetyl-CoA synthetase, myoglobin) and cellular recognition and stress genes were up-regulated (MHC1 and 2, HSP70). In the human heart, cytochrome c oxidase, acetyl-CoA synthetase, and myoglobin expression was increased in the failing heart and returned to normal with unloading. Unloading also resulted in up-regulation of HSP70. Conclusions: The genetic responses of failing human and healthy rat myocardium to mechanical unloading show similarities that appear to be independent of species differences and/or underlying disease. Thus, heterotopic heart transplantation is a relevant model for investigating the mechanisms of mechanical unloading.

References

  • 1 Frazier O H, Macris M P, Myers T J. et al . Improved survival after extended bridge to cardiac transplantation.  Ann Thorac Surg. 1994;  57 1416-1422
  • 2 Müller J, Wallukat G, Weng Y G. et al . Weaning from mechanical cardiac support in patients with idiopathic dilated cardiomyopathy.  Circul. 1997;  96 542-549
  • 3 Oz M C, Argenziano M, Catanese K A. et al . Bridge experience with long-term implantable left ventricular assist device. Are they an alternative to transplantation?.  Circul. 1997;  95 1844-1852
  • 4 Mizuno T, Weisel R D, Li R K. Reloading the heart: a new animal model of left ventricular assist device removal.  J Thorac Cardiovasc Surg. 2005;  130 99-106
  • 5 Depre C, Shipley G L, Chen W. et al . Unloaded heart in vivo replicates fetal gene expression of cardiac hypertrophy.  Nature Med. 1998;  4 1269-1275
  • 6 Klein I, Samarel A M, Welikson R. et al . Heterotopic cardiac transplantation decreases the capacity of rat myocardial protein synthesis.  Circ Res. 1991;  68 1100-1107
  • 7 Welsh D C, Dipla K, McNulty P H. et al . Preserved contractile function despite atrophic remodelling in unloaded rat hearts.  Am J Physiol Heart Circ Physiol. 2001;  281 H1131-H1136
  • 8 Doenst T, Goodwin G W, Cedars A M. et al . Load-induced changes in vivo alter substrate fluxes and insulin responsiveness of rat heart in vitro.  Metabolism. 2001;  50 1083-1090
  • 9 Diatchenko L, Lau Y C, Campbell A P. et al . Suppression subtractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and libraries.  Proc Nat Acad Sci. 1996;  93 6025-6030
  • 10 Von Stein O D, Thies W, Hofmann M. A high throughput screening for rarely transcribed differentially expressed genes.  Nucleic Acids Res. 1997;  25 2598-2602
  • 11 Wu W X, Zhang Q, Ma X H. et al . Suppression subtractive hybridization identified a marked increase in thrombospondin-1 associated with parturition in pregnant sheep myometrium.  Endocrinol. 1999;  140 2364-2371
  • 12 Ono K, Lindsey E S. Improved technique of heart transplantation in rats.  J Thorac Cardiovasc Surg. 1969;  57 225-229
  • 13 Doenst T, Schlensak C, Kobba J L. et al . A technique of heterotopic, infrarenal heart transplantation with double anastomosis in mice.  J Heart Lung Transplant. 2001;  20 762-765
  • 14 Zar J H. Biostatistical Analysis. 4th ed. Upper Saddle River; Prentice-Hall, Inc. 1999
  • 15 Razeghi P, Bruckner B A, Sharma S. et al . Mechanical unloading of the failing human heart fails to activate the protein kinase B/Akt/glycogen synthase kinase-3beta survival pathway.  Cardiol. 2003;  100 17-22
  • 16 Razeghi P, Young M E, Alcorn J L. et al . Metabolic gene expression in fetal and failing human heart.  Circul. 2001;  104 2923-2931
  • 17 Chen Y, Park S, Li Y. et al . Alterations of gene expression in failing myocardium following left ventricular assist device support.  Physiol Genomics. 2003;  14 251-260
  • 18 Napoli C, Lerman L O, Sica V. et al . Microarray analysis: a novel research tool for cardiovascular scientists and physicians.  Heart. 2003;  89 597-604
  • 19 Miller R A, Galecki A, Shmookler-Reis R J. Interpretation, design, and analysis of gene array expression experiments.  J Gerontol Ser A-Biol Sci Med Sci. 2001;  56 B52-B57
  • 20 Tsuneyoshi H, Oriyanhan W, Kanemitsu H. et al . Heterotopic transplantation of the failing rat heart as a model of left ventricular mechanical unloading toward recovery.  ASAIO J. 2005;  51 116-120
  • 21 Radovancevic B, Frazier O H, Duncan J M. Implantation technique for the HeartMate left ventricular assist device.  J Card Surg. 1992;  7 203-207
  • 22 Loschen G, Azzi A, Flohe L. Mitochondrial H2O2 formation: relationship with energy conservation.  FEBS Lett. 1973;  33 84-87
  • 23 Cadenas E. Mitochondrial free radical production and cell signalling.  Mol Aspects Med. 2004;  25 17-26
  • 24 Balaban R S, Nemoto S, Finkel T. Mitochondria, oxidants, and aging.  Cell. 2005;  120 483-495
  • 25 Taegtmeyer H, Roberts A F, Raine A E. Energy metabolism in reperfused heart muscle: metabolic correlates to return of function.  J Am Coll Cardiol. 1985;  6 864-870
  • 26 Bretschneider H J, Hubner G, Knoll D. et al . Myocardial resistance and tolerance to ischemia: physiological and chemical basis.  J Cardiovasc Surg. 1975;  16 241-260
  • 27 Fell D A. Enzymes, metabolites and fluxes.  J Exp Bot. 2005;  56 267-272
  • 28 Vogt A M, Ackermann C, Yildiz M. et al . Lactate accumulation rather than ATP depletion predicts ischemic myocardial necrosis: implications for the development of lethal myocardial injury.  Biochim Biophys Acta. 2002;  1586 219-226
  • 29 Garry D J, Kanatous S B, Mammen P P. Emerging roles for myoglobin in the heart.  Trends Cardiovasc Med. 2003;  13 111-116
  • 30 Flögel U, Laussmann T, Gödecke A. et al . Lack of myoglobin causes a switch in cardiac substrate selection.  Circ Res. 2005;  96 e68-e75
  • 31 Lepore D A, Knight K R, Anderson R L. et al . Role of priming stresses and HSP70 in protection from ischemia-reperfusion injury in cardiac and skeletal muscle.  Cell Stress Chaperones. 2001;  6 93-96
  • 32 Kelly D P, Scarpulla R C. Transcriptional regulatory circuits controlling mitochondrial biogenesis and function.  Genes Dev. 2004;  18 357-368
  • 33 Huss J M, Kelly D P. Mitochondrial energy metabolism in heart failure: a question of balance.  J Clin Invest. 2005;  115 547-555
  • 34 Stanley W C, Recchia F A, Lopaschuk G D. Myocardial substrate metabolism in the normal and failing heart.  Physiol Rev. 2005;  85 1093-1129
  • 35 Hedhli N, Pelat M, Depre C. Protein turnover in cardiac cell growth and survival.  Cardiovasc Res. 2005;  68 186-196

MD Torsten Doenst

Department of Cardiovascular Surgery
University of Freiburg

Hugstetter Straße 55

79106 Freiburg i. Br.

Germany

Phone: + 49 76 12 70 28 18

Fax: + 49 76 12 70 61 36

Email: doenst@ch11.ukl.uni-freiburg.de

    >