BIN1 is reduced and Cav1.2 trafficking is impaired in human failing cardiomyocytes

Ting Ting Hong, James W. Smyth, Kevin Y. Chu, Jacob M. Vogan, Tina S. Fong, Brian C. Jensen, Kun Fang, Marc K Halushka, Stuart D. Russell, Henry Colecraft, Charles W. Hoopes, Karen Ocorr, Neil C. Chi, Robin M. Shaw

Research output: Contribution to journalArticle

Abstract

BACKGROUND: Heart failure is a growing epidemic, and a typical aspect of heart failure pathophysiology is altered calcium transients. Normal cardiac calcium transients are initiated by Cav1.2 channels at cardiac T tubules. Bridging integrator 1 (BIN1) is a membrane scaffolding protein that causes Cav1.2 to traffic to T tubules in healthy hearts. The mechanisms of Cav1.2 trafficking in heart failure are not known. OBJECTIVE: To study BIN1 expression and its effect on Cav1.2 trafficking in failing hearts. METHODS: Intact myocardium and freshly isolated cardiomyocytes from nonfailing and end-stage failing human hearts were used to study BIN1 expression and Cav1.2 localization. To confirm Cav1.2 surface expression dependence on BIN1, patch-clamp recordings were performed of Cav1.2 current in cell lines with and without trafficking-competent BIN1. Also, in adult mouse cardiomyocytes, surface Cav1.2 and calcium transients were studied after small hairpin RNA-mediated knockdown of BIN1. For a functional readout in intact heart, calcium transients and cardiac contractility were analyzed in a zebrafish model with morpholino-mediated knockdown of BIN1. RESULTS: BIN1 expression is significantly decreased in failing cardiomyocytes at both mRNA (30% down) and protein (36% down) levels. Peripheral Cav1.2 is reduced to 42% by imaging, and a biochemical T-tubule fraction of Cav1.2 is reduced to 68%. The total calcium current is reduced to 41% in a cell line expressing a nontrafficking BIN1 mutant. In mouse cardiomyocytes, BIN1 knockdown decreases surface Cav1.2 and impairs calcium transients. In zebrafish hearts, BIN1 knockdown causes a 75% reduction in calcium transients and severe ventricular contractile dysfunction. CONCLUSIONS: The data indicate that BIN1 is significantly reduced in human heart failure, and this reduction impairs Cav1.2 trafficking, calcium transients, and contractility.

Original languageEnglish (US)
Pages (from-to)812-820
Number of pages9
JournalHeart Rhythm
Volume9
Issue number5
DOIs
StatePublished - May 2012

Fingerprint

Cardiac Myocytes
Calcium
Heart Failure
Zebrafish
Ventricular Dysfunction
Cell Line
Morpholinos
Small Interfering RNA
Myocardium
Membrane Proteins
Messenger RNA
Proteins

Keywords

  • Calcium
  • Calcium transient
  • Cardiomyopathy
  • Heart failure
  • Ion channels
  • L-type calcium channel
  • Trafficking

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)

Cite this

Hong, T. T., Smyth, J. W., Chu, K. Y., Vogan, J. M., Fong, T. S., Jensen, B. C., ... Shaw, R. M. (2012). BIN1 is reduced and Cav1.2 trafficking is impaired in human failing cardiomyocytes. Heart Rhythm, 9(5), 812-820. https://doi.org/10.1016/j.hrthm.2011.11.055

BIN1 is reduced and Cav1.2 trafficking is impaired in human failing cardiomyocytes. / Hong, Ting Ting; Smyth, James W.; Chu, Kevin Y.; Vogan, Jacob M.; Fong, Tina S.; Jensen, Brian C.; Fang, Kun; Halushka, Marc K; Russell, Stuart D.; Colecraft, Henry; Hoopes, Charles W.; Ocorr, Karen; Chi, Neil C.; Shaw, Robin M.

In: Heart Rhythm, Vol. 9, No. 5, 05.2012, p. 812-820.

Research output: Contribution to journalArticle

Hong, TT, Smyth, JW, Chu, KY, Vogan, JM, Fong, TS, Jensen, BC, Fang, K, Halushka, MK, Russell, SD, Colecraft, H, Hoopes, CW, Ocorr, K, Chi, NC & Shaw, RM 2012, 'BIN1 is reduced and Cav1.2 trafficking is impaired in human failing cardiomyocytes', Heart Rhythm, vol. 9, no. 5, pp. 812-820. https://doi.org/10.1016/j.hrthm.2011.11.055
Hong, Ting Ting ; Smyth, James W. ; Chu, Kevin Y. ; Vogan, Jacob M. ; Fong, Tina S. ; Jensen, Brian C. ; Fang, Kun ; Halushka, Marc K ; Russell, Stuart D. ; Colecraft, Henry ; Hoopes, Charles W. ; Ocorr, Karen ; Chi, Neil C. ; Shaw, Robin M. / BIN1 is reduced and Cav1.2 trafficking is impaired in human failing cardiomyocytes. In: Heart Rhythm. 2012 ; Vol. 9, No. 5. pp. 812-820.
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abstract = "BACKGROUND: Heart failure is a growing epidemic, and a typical aspect of heart failure pathophysiology is altered calcium transients. Normal cardiac calcium transients are initiated by Cav1.2 channels at cardiac T tubules. Bridging integrator 1 (BIN1) is a membrane scaffolding protein that causes Cav1.2 to traffic to T tubules in healthy hearts. The mechanisms of Cav1.2 trafficking in heart failure are not known. OBJECTIVE: To study BIN1 expression and its effect on Cav1.2 trafficking in failing hearts. METHODS: Intact myocardium and freshly isolated cardiomyocytes from nonfailing and end-stage failing human hearts were used to study BIN1 expression and Cav1.2 localization. To confirm Cav1.2 surface expression dependence on BIN1, patch-clamp recordings were performed of Cav1.2 current in cell lines with and without trafficking-competent BIN1. Also, in adult mouse cardiomyocytes, surface Cav1.2 and calcium transients were studied after small hairpin RNA-mediated knockdown of BIN1. For a functional readout in intact heart, calcium transients and cardiac contractility were analyzed in a zebrafish model with morpholino-mediated knockdown of BIN1. RESULTS: BIN1 expression is significantly decreased in failing cardiomyocytes at both mRNA (30{\%} down) and protein (36{\%} down) levels. Peripheral Cav1.2 is reduced to 42{\%} by imaging, and a biochemical T-tubule fraction of Cav1.2 is reduced to 68{\%}. The total calcium current is reduced to 41{\%} in a cell line expressing a nontrafficking BIN1 mutant. In mouse cardiomyocytes, BIN1 knockdown decreases surface Cav1.2 and impairs calcium transients. In zebrafish hearts, BIN1 knockdown causes a 75{\%} reduction in calcium transients and severe ventricular contractile dysfunction. CONCLUSIONS: The data indicate that BIN1 is significantly reduced in human heart failure, and this reduction impairs Cav1.2 trafficking, calcium transients, and contractility.",
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AU - Hong, Ting Ting

AU - Smyth, James W.

AU - Chu, Kevin Y.

AU - Vogan, Jacob M.

AU - Fong, Tina S.

AU - Jensen, Brian C.

AU - Fang, Kun

AU - Halushka, Marc K

AU - Russell, Stuart D.

AU - Colecraft, Henry

AU - Hoopes, Charles W.

AU - Ocorr, Karen

AU - Chi, Neil C.

AU - Shaw, Robin M.

PY - 2012/5

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N2 - BACKGROUND: Heart failure is a growing epidemic, and a typical aspect of heart failure pathophysiology is altered calcium transients. Normal cardiac calcium transients are initiated by Cav1.2 channels at cardiac T tubules. Bridging integrator 1 (BIN1) is a membrane scaffolding protein that causes Cav1.2 to traffic to T tubules in healthy hearts. The mechanisms of Cav1.2 trafficking in heart failure are not known. OBJECTIVE: To study BIN1 expression and its effect on Cav1.2 trafficking in failing hearts. METHODS: Intact myocardium and freshly isolated cardiomyocytes from nonfailing and end-stage failing human hearts were used to study BIN1 expression and Cav1.2 localization. To confirm Cav1.2 surface expression dependence on BIN1, patch-clamp recordings were performed of Cav1.2 current in cell lines with and without trafficking-competent BIN1. Also, in adult mouse cardiomyocytes, surface Cav1.2 and calcium transients were studied after small hairpin RNA-mediated knockdown of BIN1. For a functional readout in intact heart, calcium transients and cardiac contractility were analyzed in a zebrafish model with morpholino-mediated knockdown of BIN1. RESULTS: BIN1 expression is significantly decreased in failing cardiomyocytes at both mRNA (30% down) and protein (36% down) levels. Peripheral Cav1.2 is reduced to 42% by imaging, and a biochemical T-tubule fraction of Cav1.2 is reduced to 68%. The total calcium current is reduced to 41% in a cell line expressing a nontrafficking BIN1 mutant. In mouse cardiomyocytes, BIN1 knockdown decreases surface Cav1.2 and impairs calcium transients. In zebrafish hearts, BIN1 knockdown causes a 75% reduction in calcium transients and severe ventricular contractile dysfunction. CONCLUSIONS: The data indicate that BIN1 is significantly reduced in human heart failure, and this reduction impairs Cav1.2 trafficking, calcium transients, and contractility.

AB - BACKGROUND: Heart failure is a growing epidemic, and a typical aspect of heart failure pathophysiology is altered calcium transients. Normal cardiac calcium transients are initiated by Cav1.2 channels at cardiac T tubules. Bridging integrator 1 (BIN1) is a membrane scaffolding protein that causes Cav1.2 to traffic to T tubules in healthy hearts. The mechanisms of Cav1.2 trafficking in heart failure are not known. OBJECTIVE: To study BIN1 expression and its effect on Cav1.2 trafficking in failing hearts. METHODS: Intact myocardium and freshly isolated cardiomyocytes from nonfailing and end-stage failing human hearts were used to study BIN1 expression and Cav1.2 localization. To confirm Cav1.2 surface expression dependence on BIN1, patch-clamp recordings were performed of Cav1.2 current in cell lines with and without trafficking-competent BIN1. Also, in adult mouse cardiomyocytes, surface Cav1.2 and calcium transients were studied after small hairpin RNA-mediated knockdown of BIN1. For a functional readout in intact heart, calcium transients and cardiac contractility were analyzed in a zebrafish model with morpholino-mediated knockdown of BIN1. RESULTS: BIN1 expression is significantly decreased in failing cardiomyocytes at both mRNA (30% down) and protein (36% down) levels. Peripheral Cav1.2 is reduced to 42% by imaging, and a biochemical T-tubule fraction of Cav1.2 is reduced to 68%. The total calcium current is reduced to 41% in a cell line expressing a nontrafficking BIN1 mutant. In mouse cardiomyocytes, BIN1 knockdown decreases surface Cav1.2 and impairs calcium transients. In zebrafish hearts, BIN1 knockdown causes a 75% reduction in calcium transients and severe ventricular contractile dysfunction. CONCLUSIONS: The data indicate that BIN1 is significantly reduced in human heart failure, and this reduction impairs Cav1.2 trafficking, calcium transients, and contractility.

KW - Calcium

KW - Calcium transient

KW - Cardiomyopathy

KW - Heart failure

KW - Ion channels

KW - L-type calcium channel

KW - Trafficking

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