Impaired diastolic function after exchange of endogenous troponin I with C-terminal truncated troponin I in human cardiac muscle

Nadiya A. Narolska, Nicoletta Piroddi, Alexandra Belus, Nicky M. Boontje, Beatrice Scellini, Sascha Deppermann, Ruud Zaremba, Rene J. Musters, Cris Dos Remedios, Kornelia Jaquet, D. Brian Foster, Anne M. Murphy, Jennifer E. Van Eyk, Chiara Tesi, Corrado Poggesi, Jolanda Van Der Velden, Ger J.M. Stienen

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

The specific and selective proteolysis of cardiac troponin I (cTnI) has been proposed to play a key role in human ischemic myocardial disease, including stunning and acute pressure overload. In this study, the functional implications of cTnI proteolysis were investigated in human cardiac tissue for the first time. The predominant human cTnI degradation product (cTnI1-192) and full-length cTnI were expressed in Escherichia coli, purified, reconstituted with the other cardiac troponin subunits, troponin T and C, and subsequently exchanged into human cardiac myofibrils and permeabilized cardiomyocytes isolated from healthy donor hearts. Maximal isometric force and kinetic parameters were measured in myofibrils, using rapid solution switching, whereas force development was measured in single cardiomyocytes at various calcium concentrations, at sarcomere lengths of 1.9 and 2.2 μm, and after treatment with the catalytic subunit of protein kinase A (PKA) to mimic β-adrenergic stimulation. One-dimensional gel electrophoresis, Western immunoblotting, and 3D imaging revealed that approximately 50% of endogenous cTnI had been homogeneously replaced by cTnI1-192 in both myofibrils and cardiomyocytes. Maximal tension was not affected, whereas the rates of force activation and redevelopment as well as relaxation kinetics were slowed down. Ca sensitivity of the contractile apparatus was increased in preparations containing cTnI1-192 (pCa50: 5.73±0.03 versus 5.52±0.03 for cTnI1-192 and full-length cTnI, respectively). The sarcomere length dependency of force development and the desensitizing effect of PKA were preserved in cTnI1-192-exchanged cardiomyocytes. These results indicate that degradation of cTnI in human myocardium may impair diastolic function, whereas systolic function is largely preserved.

Original languageEnglish
Pages (from-to)1012-1020
Number of pages9
JournalCirculation Research
Volume99
Issue number9
DOIs
Publication statusPublished - 1 Oct 2006

Cite this

Narolska, N. A., Piroddi, N., Belus, A., Boontje, N. M., Scellini, B., Deppermann, S., ... Stienen, G. J. M. (2006). Impaired diastolic function after exchange of endogenous troponin I with C-terminal truncated troponin I in human cardiac muscle. Circulation Research, 99(9), 1012-1020. https://doi.org/10.1161/01.RES.0000248753.30340.af
Narolska, Nadiya A. ; Piroddi, Nicoletta ; Belus, Alexandra ; Boontje, Nicky M. ; Scellini, Beatrice ; Deppermann, Sascha ; Zaremba, Ruud ; Musters, Rene J. ; Dos Remedios, Cris ; Jaquet, Kornelia ; Foster, D. Brian ; Murphy, Anne M. ; Van Eyk, Jennifer E. ; Tesi, Chiara ; Poggesi, Corrado ; Van Der Velden, Jolanda ; Stienen, Ger J.M. / Impaired diastolic function after exchange of endogenous troponin I with C-terminal truncated troponin I in human cardiac muscle. In: Circulation Research. 2006 ; Vol. 99, No. 9. pp. 1012-1020.
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abstract = "The specific and selective proteolysis of cardiac troponin I (cTnI) has been proposed to play a key role in human ischemic myocardial disease, including stunning and acute pressure overload. In this study, the functional implications of cTnI proteolysis were investigated in human cardiac tissue for the first time. The predominant human cTnI degradation product (cTnI1-192) and full-length cTnI were expressed in Escherichia coli, purified, reconstituted with the other cardiac troponin subunits, troponin T and C, and subsequently exchanged into human cardiac myofibrils and permeabilized cardiomyocytes isolated from healthy donor hearts. Maximal isometric force and kinetic parameters were measured in myofibrils, using rapid solution switching, whereas force development was measured in single cardiomyocytes at various calcium concentrations, at sarcomere lengths of 1.9 and 2.2 μm, and after treatment with the catalytic subunit of protein kinase A (PKA) to mimic β-adrenergic stimulation. One-dimensional gel electrophoresis, Western immunoblotting, and 3D imaging revealed that approximately 50{\%} of endogenous cTnI had been homogeneously replaced by cTnI1-192 in both myofibrils and cardiomyocytes. Maximal tension was not affected, whereas the rates of force activation and redevelopment as well as relaxation kinetics were slowed down. Ca sensitivity of the contractile apparatus was increased in preparations containing cTnI1-192 (pCa50: 5.73±0.03 versus 5.52±0.03 for cTnI1-192 and full-length cTnI, respectively). The sarcomere length dependency of force development and the desensitizing effect of PKA were preserved in cTnI1-192-exchanged cardiomyocytes. These results indicate that degradation of cTnI in human myocardium may impair diastolic function, whereas systolic function is largely preserved.",
keywords = "Cardiac function, Cardiomyocytes, Cardiomyopathy, Contractility, Ischemia",
author = "Narolska, {Nadiya A.} and Nicoletta Piroddi and Alexandra Belus and Boontje, {Nicky M.} and Beatrice Scellini and Sascha Deppermann and Ruud Zaremba and Musters, {Rene J.} and {Dos Remedios}, Cris and Kornelia Jaquet and Foster, {D. Brian} and Murphy, {Anne M.} and {Van Eyk}, {Jennifer E.} and Chiara Tesi and Corrado Poggesi and {Van Der Velden}, Jolanda and Stienen, {Ger J.M.}",
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Narolska, NA, Piroddi, N, Belus, A, Boontje, NM, Scellini, B, Deppermann, S, Zaremba, R, Musters, RJ, Dos Remedios, C, Jaquet, K, Foster, DB, Murphy, AM, Van Eyk, JE, Tesi, C, Poggesi, C, Van Der Velden, J & Stienen, GJM 2006, 'Impaired diastolic function after exchange of endogenous troponin I with C-terminal truncated troponin I in human cardiac muscle' Circulation Research, vol. 99, no. 9, pp. 1012-1020. https://doi.org/10.1161/01.RES.0000248753.30340.af

Impaired diastolic function after exchange of endogenous troponin I with C-terminal truncated troponin I in human cardiac muscle. / Narolska, Nadiya A.; Piroddi, Nicoletta; Belus, Alexandra; Boontje, Nicky M.; Scellini, Beatrice; Deppermann, Sascha; Zaremba, Ruud; Musters, Rene J.; Dos Remedios, Cris; Jaquet, Kornelia; Foster, D. Brian; Murphy, Anne M.; Van Eyk, Jennifer E.; Tesi, Chiara; Poggesi, Corrado; Van Der Velden, Jolanda; Stienen, Ger J.M.

In: Circulation Research, Vol. 99, No. 9, 01.10.2006, p. 1012-1020.

Research output: Contribution to journalArticleAcademicpeer-review

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T1 - Impaired diastolic function after exchange of endogenous troponin I with C-terminal truncated troponin I in human cardiac muscle

AU - Narolska, Nadiya A.

AU - Piroddi, Nicoletta

AU - Belus, Alexandra

AU - Boontje, Nicky M.

AU - Scellini, Beatrice

AU - Deppermann, Sascha

AU - Zaremba, Ruud

AU - Musters, Rene J.

AU - Dos Remedios, Cris

AU - Jaquet, Kornelia

AU - Foster, D. Brian

AU - Murphy, Anne M.

AU - Van Eyk, Jennifer E.

AU - Tesi, Chiara

AU - Poggesi, Corrado

AU - Van Der Velden, Jolanda

AU - Stienen, Ger J.M.

PY - 2006/10/1

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N2 - The specific and selective proteolysis of cardiac troponin I (cTnI) has been proposed to play a key role in human ischemic myocardial disease, including stunning and acute pressure overload. In this study, the functional implications of cTnI proteolysis were investigated in human cardiac tissue for the first time. The predominant human cTnI degradation product (cTnI1-192) and full-length cTnI were expressed in Escherichia coli, purified, reconstituted with the other cardiac troponin subunits, troponin T and C, and subsequently exchanged into human cardiac myofibrils and permeabilized cardiomyocytes isolated from healthy donor hearts. Maximal isometric force and kinetic parameters were measured in myofibrils, using rapid solution switching, whereas force development was measured in single cardiomyocytes at various calcium concentrations, at sarcomere lengths of 1.9 and 2.2 μm, and after treatment with the catalytic subunit of protein kinase A (PKA) to mimic β-adrenergic stimulation. One-dimensional gel electrophoresis, Western immunoblotting, and 3D imaging revealed that approximately 50% of endogenous cTnI had been homogeneously replaced by cTnI1-192 in both myofibrils and cardiomyocytes. Maximal tension was not affected, whereas the rates of force activation and redevelopment as well as relaxation kinetics were slowed down. Ca sensitivity of the contractile apparatus was increased in preparations containing cTnI1-192 (pCa50: 5.73±0.03 versus 5.52±0.03 for cTnI1-192 and full-length cTnI, respectively). The sarcomere length dependency of force development and the desensitizing effect of PKA were preserved in cTnI1-192-exchanged cardiomyocytes. These results indicate that degradation of cTnI in human myocardium may impair diastolic function, whereas systolic function is largely preserved.

AB - The specific and selective proteolysis of cardiac troponin I (cTnI) has been proposed to play a key role in human ischemic myocardial disease, including stunning and acute pressure overload. In this study, the functional implications of cTnI proteolysis were investigated in human cardiac tissue for the first time. The predominant human cTnI degradation product (cTnI1-192) and full-length cTnI were expressed in Escherichia coli, purified, reconstituted with the other cardiac troponin subunits, troponin T and C, and subsequently exchanged into human cardiac myofibrils and permeabilized cardiomyocytes isolated from healthy donor hearts. Maximal isometric force and kinetic parameters were measured in myofibrils, using rapid solution switching, whereas force development was measured in single cardiomyocytes at various calcium concentrations, at sarcomere lengths of 1.9 and 2.2 μm, and after treatment with the catalytic subunit of protein kinase A (PKA) to mimic β-adrenergic stimulation. One-dimensional gel electrophoresis, Western immunoblotting, and 3D imaging revealed that approximately 50% of endogenous cTnI had been homogeneously replaced by cTnI1-192 in both myofibrils and cardiomyocytes. Maximal tension was not affected, whereas the rates of force activation and redevelopment as well as relaxation kinetics were slowed down. Ca sensitivity of the contractile apparatus was increased in preparations containing cTnI1-192 (pCa50: 5.73±0.03 versus 5.52±0.03 for cTnI1-192 and full-length cTnI, respectively). The sarcomere length dependency of force development and the desensitizing effect of PKA were preserved in cTnI1-192-exchanged cardiomyocytes. These results indicate that degradation of cTnI in human myocardium may impair diastolic function, whereas systolic function is largely preserved.

KW - Cardiac function

KW - Cardiomyocytes

KW - Cardiomyopathy

KW - Contractility

KW - Ischemia

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