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Histidine button engineered into cardiac troponin I protects the ischemic and failing heart

Nature Medicine volume 12, pages 181–189 (2006)Cite this article

Abstract

The myofilament protein troponin I (TnI) has a key isoform-dependent role in the development of contractile failure during acidosis and ischemia. Here we show that cardiac performance in vitro and in vivo is enhanced when a single histidine residue present in the fetal cardiac TnI isoform is substituted into the adult cardiac TnI isoform at codon 164. The most marked effects are observed under the acute challenges of acidosis, hypoxia, ischemia and ischemia-reperfusion, in chronic heart failure in transgenic mice and in myocytes from failing human hearts. In the isolated heart, histidine-modified TnI improves systolic and diastolic function and mitigates reperfusion-associated ventricular arrhythmias. Cardiac performance is markedly enhanced in transgenic hearts during reperfusion despite a high-energy phosphate content similar to that in nontransgenic hearts, providing evidence for greater energetic economy. This pH-sensitive 'histidine button' engineered in TnI produces a titratable molecular switch that 'senses' changes in the intracellular milieu of the cardiac myocyte and responds by preferentially augmenting acute and long-term function under pathophysiological conditions. Myofilament-based inotropy may represent a therapeutic avenue to improve myocardial performance in the ischemic and failing heart.

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Figure 1: cTnI A164H transgene construct and expression, and isometric tension in isolated single mouse cardiac myocytes.
Figure 2: Acidic pH challenge in single intact myocytes and isolated hearts.
Figure 3: In vivo micromanometry measurements under hypoxic and ischemic challenges.
Figure 4: Hemodynamics, arrhythmias and energetics during ischemia and reperfusion in isolated hearts.
Figure 5: Assessment of function in the chronically failing mouse heart in vivo and in human failing cardiac myocytes ex vivo.

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Acknowledgements

We thank J. Li, A. Lopatin, S. Whitesall, R. Liao, K. Converso, M. Russell, F. Pagani, H. Patel, E. DeVaney, D. Dyke, E. Favre, A. Sober-Rankin, K. Pasyk and S. Forfa for assistance and expertise. This work was supported by grants from the American Heart Association (0475032N to S.M.D.) and the US National Institutes of Health (HL67254 to M.V.W., HL52320 and HL63985 to J.S.I., and HL059301 to J.M.M.).

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Author notes

  1. Sharlene M Day and Margaret V Westfall: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Internal Medicine, 1150 West Medical Center Drive, 7301 MSRB III, University of Michigan Medical School, Ann Arbor, 48109, Michigan, USA

    Sharlene M Day & Joseph M Metzger

  2. Department of Cardiac Surgery, 1150 West Medical Center Drive, B570D MSRB II, University of Michigan Medical School, Ann Arbor, 48109, Michigan, USA

    Margaret V Westfall

  3. Department of Molecular and Integrative Physiology, 1301 East Catherine Street, 7730 Medical Science II, University of Michigan Medical School, Ann Arbor, 48109, Michigan, USA

    Margaret V Westfall, Ekaterina V Fomicheva, Soichiro Yasuda, Nathan C La Cross, Louis G D'Alecy & Joseph M Metzger

  4. Department of Medicine, NMR Laboratory for Physiological Chemistry, Brigham and Women's Hospital, Harvard Medical School, 221 Longwood Avenue, Boston, 02115, Massachusetts, USA

    Kirsten Hoyer & Joanne S Ingwall

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Supplementary information

Supplementary Fig. 1

Representative tension-pCa relationships in adult single cardiac myocytes expressing cTnI (control), ssTnI or ssTnI H132A. (PDF 770 kb)

Supplementary Fig. 2

Summary of pCa50 in cardiac myocytes after cTnI A164H adenoviral gene transfer in isolated rat cardiac myocytes. (PDF 25 kb)

Supplementary Fig. 3

Confocal imaging of indirect immunofluorescently labeled myocardial cryosections. (PDF 386 kb)

Supplementary Fig. 4

Cell and organ morphometrics, histology and electron microscopy. (PDF 145 kb)

Supplementary Fig. 5

Protein expression in intact hearts. (PDF 54 kb)

Supplementary Fig. 6

Isolated myocyte and heart organ responses to β-adrenergic stimuli. (PDF 605 kb)

Supplementary Fig. 7

Intraventricular pressures during ischemia and reperfusion in isolated mouse hearts from two independent transgenic mouse lines. (PDF 40 kb)

Supplementary Fig. 8

Effects of ischemia and reperfusion in cTnI A164H mice in vivo. (PDF 25 kb)

Supplementary Table 1

Energetic enzyme measurements. (PDF 10 kb)

Supplementary Methods (PDF 50 kb)

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Day, S., Westfall, M., Fomicheva, E. et al. Histidine button engineered into cardiac troponin I protects the ischemic and failing heart. Nat Med 12, 181–189 (2006). https://doi.org/10.1038/nm1346

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