Targeting miR-423-5p Reverses Exercise Training-Induced HCN4 Channel Remodeling and Sinus Bradycardia

Alicia D'Souza; Charles M. Pearman; Yanwen Wang; Shu Nakao; Sunil Jit R.J. Logantha; Charlotte Cox; Hayley Bennett; Yu Zhang; Anne Berit Johnsen; Nora Linscheid; Pi Camilla Poulsen; Jonathan Elliott; Jessica Coulson; Jamie McPhee; Abigail Robertson; Paula A. Da Costa Martins; Ashraf Kitmitto; Ulrik Wisløff; Elizabeth J. Cartwright; Oliver Monfredi; Alicia Lundby; Halina Dobrzynski; Delvac Oceandy; Gwilym M. Morris; Mark R. Boyett

doi:10.1161/CIRCRESAHA.117.311607

Targeting miR-423-5p Reverses Exercise Training-Induced HCN4 Channel Remodeling and Sinus Bradycardia

Alicia D'Souza, Charles M. Pearman, Yanwen Wang, Shu Nakao, Sunil Jit R.J. Logantha, Charlotte Cox, Hayley Bennett, Yu Zhang, Anne Berit Johnsen, Nora Linscheid, Pi Camilla Poulsen, Jonathan Elliott, Jessica Coulson, Jamie McPhee, Abigail Robertson, Paula A. Da Costa Martins, Ashraf Kitmitto, Ulrik Wisløff, Elizabeth J. Cartwright, Oliver MonfrediAlicia Lundby, Halina Dobrzynski, Delvac Oceandy, Gwilym M. Morris, Mark R. Boyett^*

^*Corresponding author for this work

31 Citations (Scopus)

126 Downloads (Pure)

Abstract

Rationale: Downregulation of the pacemaking ion channel, HCN4 (hyperpolarization-activated cyclic nucleotide gated channel 4), and the corresponding ionic current, I_f, underlies exercise training-induced sinus bradycardia in rodents. If this occurs in humans, it could explain the increased incidence of bradyarrhythmias in veteran athletes, and it will be important to understand the underlying processes. Objective: To test the role of HCN4 in the training-induced bradycardia in human athletes and investigate the role of microRNAs (miRs) in the repression of HCN4. Methods and Results: As in rodents, the intrinsic heart rate was significantly lower in human athletes than in nonathletes, and in all subjects, the rate-lowering effect of the HCN selective blocker, ivabradine, was significantly correlated with the intrinsic heart rate, consistent with HCN repression in athletes. Next-generation sequencing and quantitative real-time reverse transcription polymerase chain reaction showed remodeling of miRs in the sinus node of swim-trained mice. Computational predictions highlighted a prominent role for miR-423-5p. Interaction between miR-423-5p and HCN4 was confirmed by a dose-dependent reduction in HCN4 3′-untranslated region luciferase reporter activity on cotransfection with precursor miR-423-5p (abolished by mutation of predicted recognition elements). Knockdown of miR-423-5p with anti-miR-423-5p reversed training-induced bradycardia via rescue of HCN4 and I_f. Further experiments showed that in the sinus node of swim-trained mice, upregulation of miR-423-5p (intronic miR) and its host gene, NSRP1, is driven by an upregulation of the transcription factor Nkx2.5. Conclusions: HCN remodeling likely occurs in human athletes, as well as in rodent models. miR-423-5p contributes to training-induced bradycardia by targeting HCN4. This work presents the first evidence of miR control of HCN4 and heart rate. miR-423-5p could be a therapeutic target for pathological sinus node dysfunction in veteran athletes.

Original language	English
Journal	Circulation Research
Volume	121
Issue number	9
Pages (from-to)	1058-1068
Number of pages	11
ISSN	0009-7330
DOIs	https://doi.org/10.1161/CIRCRESAHA.117.311607
Publication status	Published - Oct 2017

Keywords

athletes
exercise training
ion channel remodeling
micro-RNAs
sinoatrial node
sinus bradycardia

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Targeting miR-423-5p Reverses Exercise Training–Induced HCN4 Channel Remodeling and Sinus BradycardiaFinal published version, 4.88 MBLicence: CC BY

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D'Souza, A., Pearman, C. M., Wang, Y., Nakao, S., Logantha, S. J. R. J., Cox, C., Bennett, H., Zhang, Y., Johnsen, A. B., Linscheid, N., Poulsen, P. C., Elliott, J., Coulson, J., McPhee, J., Robertson, A., Da Costa Martins, P. A., Kitmitto, A., Wisløff, U., Cartwright, E. J., ... Boyett, M. R. (2017). Targeting miR-423-5p Reverses Exercise Training-Induced HCN4 Channel Remodeling and Sinus Bradycardia. Circulation Research, 121(9), 1058-1068. https://doi.org/10.1161/CIRCRESAHA.117.311607

D'Souza, A, Pearman, CM, Wang, Y, Nakao, S, Logantha, SJRJ, Cox, C, Bennett, H, Zhang, Y, Johnsen, AB, Linscheid, N, Poulsen, PC, Elliott, J, Coulson, J, McPhee, J, Robertson, A, Da Costa Martins, PA, Kitmitto, A, Wisløff, U, Cartwright, EJ, Monfredi, O, Lundby, A, Dobrzynski, H, Oceandy, D, Morris, GM & Boyett, MR 2017, 'Targeting miR-423-5p Reverses Exercise Training-Induced HCN4 Channel Remodeling and Sinus Bradycardia', Circulation Research, vol. 121, no. 9, pp. 1058-1068. https://doi.org/10.1161/CIRCRESAHA.117.311607

@article{5593149643a84054bb62edb8b678187a,

title = "Targeting miR-423-5p Reverses Exercise Training-Induced HCN4 Channel Remodeling and Sinus Bradycardia",

abstract = "Rationale: Downregulation of the pacemaking ion channel, HCN4 (hyperpolarization-activated cyclic nucleotide gated channel 4), and the corresponding ionic current, If, underlies exercise training-induced sinus bradycardia in rodents. If this occurs in humans, it could explain the increased incidence of bradyarrhythmias in veteran athletes, and it will be important to understand the underlying processes. Objective: To test the role of HCN4 in the training-induced bradycardia in human athletes and investigate the role of microRNAs (miRs) in the repression of HCN4. Methods and Results: As in rodents, the intrinsic heart rate was significantly lower in human athletes than in nonathletes, and in all subjects, the rate-lowering effect of the HCN selective blocker, ivabradine, was significantly correlated with the intrinsic heart rate, consistent with HCN repression in athletes. Next-generation sequencing and quantitative real-time reverse transcription polymerase chain reaction showed remodeling of miRs in the sinus node of swim-trained mice. Computational predictions highlighted a prominent role for miR-423-5p. Interaction between miR-423-5p and HCN4 was confirmed by a dose-dependent reduction in HCN4 3′-untranslated region luciferase reporter activity on cotransfection with precursor miR-423-5p (abolished by mutation of predicted recognition elements). Knockdown of miR-423-5p with anti-miR-423-5p reversed training-induced bradycardia via rescue of HCN4 and If. Further experiments showed that in the sinus node of swim-trained mice, upregulation of miR-423-5p (intronic miR) and its host gene, NSRP1, is driven by an upregulation of the transcription factor Nkx2.5. Conclusions: HCN remodeling likely occurs in human athletes, as well as in rodent models. miR-423-5p contributes to training-induced bradycardia by targeting HCN4. This work presents the first evidence of miR control of HCN4 and heart rate. miR-423-5p could be a therapeutic target for pathological sinus node dysfunction in veteran athletes.",

keywords = "athletes, exercise training, ion channel remodeling, micro-RNAs, sinoatrial node, sinus bradycardia",

author = "Alicia D'Souza and Pearman, {Charles M.} and Yanwen Wang and Shu Nakao and Logantha, {Sunil Jit R.J.} and Charlotte Cox and Hayley Bennett and Yu Zhang and Johnsen, {Anne Berit} and Nora Linscheid and Poulsen, {Pi Camilla} and Jonathan Elliott and Jessica Coulson and Jamie McPhee and Abigail Robertson and {Da Costa Martins}, {Paula A.} and Ashraf Kitmitto and Ulrik Wisl{\o}ff and Cartwright, {Elizabeth J.} and Oliver Monfredi and Alicia Lundby and Halina Dobrzynski and Delvac Oceandy and Morris, {Gwilym M.} and Boyett, {Mark R.}",

year = "2017",

month = oct,

doi = "10.1161/CIRCRESAHA.117.311607",

language = "English",

volume = "121",

pages = "1058--1068",

journal = "Circulation",

issn = "0009-7322",

publisher = "AHA/ASA",

number = "9",

}

TY - JOUR

T1 - Targeting miR-423-5p Reverses Exercise Training-Induced HCN4 Channel Remodeling and Sinus Bradycardia

AU - D'Souza, Alicia

AU - Pearman, Charles M.

AU - Wang, Yanwen

AU - Nakao, Shu

AU - Logantha, Sunil Jit R.J.

AU - Cox, Charlotte

AU - Bennett, Hayley

AU - Zhang, Yu

AU - Johnsen, Anne Berit

AU - Linscheid, Nora

AU - Poulsen, Pi Camilla

AU - Elliott, Jonathan

AU - Coulson, Jessica

AU - McPhee, Jamie

AU - Robertson, Abigail

AU - Da Costa Martins, Paula A.

AU - Kitmitto, Ashraf

AU - Wisløff, Ulrik

AU - Cartwright, Elizabeth J.

AU - Monfredi, Oliver

AU - Lundby, Alicia

AU - Dobrzynski, Halina

AU - Oceandy, Delvac

AU - Morris, Gwilym M.

AU - Boyett, Mark R.

PY - 2017/10

Y1 - 2017/10

N2 - Rationale: Downregulation of the pacemaking ion channel, HCN4 (hyperpolarization-activated cyclic nucleotide gated channel 4), and the corresponding ionic current, If, underlies exercise training-induced sinus bradycardia in rodents. If this occurs in humans, it could explain the increased incidence of bradyarrhythmias in veteran athletes, and it will be important to understand the underlying processes. Objective: To test the role of HCN4 in the training-induced bradycardia in human athletes and investigate the role of microRNAs (miRs) in the repression of HCN4. Methods and Results: As in rodents, the intrinsic heart rate was significantly lower in human athletes than in nonathletes, and in all subjects, the rate-lowering effect of the HCN selective blocker, ivabradine, was significantly correlated with the intrinsic heart rate, consistent with HCN repression in athletes. Next-generation sequencing and quantitative real-time reverse transcription polymerase chain reaction showed remodeling of miRs in the sinus node of swim-trained mice. Computational predictions highlighted a prominent role for miR-423-5p. Interaction between miR-423-5p and HCN4 was confirmed by a dose-dependent reduction in HCN4 3′-untranslated region luciferase reporter activity on cotransfection with precursor miR-423-5p (abolished by mutation of predicted recognition elements). Knockdown of miR-423-5p with anti-miR-423-5p reversed training-induced bradycardia via rescue of HCN4 and If. Further experiments showed that in the sinus node of swim-trained mice, upregulation of miR-423-5p (intronic miR) and its host gene, NSRP1, is driven by an upregulation of the transcription factor Nkx2.5. Conclusions: HCN remodeling likely occurs in human athletes, as well as in rodent models. miR-423-5p contributes to training-induced bradycardia by targeting HCN4. This work presents the first evidence of miR control of HCN4 and heart rate. miR-423-5p could be a therapeutic target for pathological sinus node dysfunction in veteran athletes.

AB - Rationale: Downregulation of the pacemaking ion channel, HCN4 (hyperpolarization-activated cyclic nucleotide gated channel 4), and the corresponding ionic current, If, underlies exercise training-induced sinus bradycardia in rodents. If this occurs in humans, it could explain the increased incidence of bradyarrhythmias in veteran athletes, and it will be important to understand the underlying processes. Objective: To test the role of HCN4 in the training-induced bradycardia in human athletes and investigate the role of microRNAs (miRs) in the repression of HCN4. Methods and Results: As in rodents, the intrinsic heart rate was significantly lower in human athletes than in nonathletes, and in all subjects, the rate-lowering effect of the HCN selective blocker, ivabradine, was significantly correlated with the intrinsic heart rate, consistent with HCN repression in athletes. Next-generation sequencing and quantitative real-time reverse transcription polymerase chain reaction showed remodeling of miRs in the sinus node of swim-trained mice. Computational predictions highlighted a prominent role for miR-423-5p. Interaction between miR-423-5p and HCN4 was confirmed by a dose-dependent reduction in HCN4 3′-untranslated region luciferase reporter activity on cotransfection with precursor miR-423-5p (abolished by mutation of predicted recognition elements). Knockdown of miR-423-5p with anti-miR-423-5p reversed training-induced bradycardia via rescue of HCN4 and If. Further experiments showed that in the sinus node of swim-trained mice, upregulation of miR-423-5p (intronic miR) and its host gene, NSRP1, is driven by an upregulation of the transcription factor Nkx2.5. Conclusions: HCN remodeling likely occurs in human athletes, as well as in rodent models. miR-423-5p contributes to training-induced bradycardia by targeting HCN4. This work presents the first evidence of miR control of HCN4 and heart rate. miR-423-5p could be a therapeutic target for pathological sinus node dysfunction in veteran athletes.

KW - athletes

KW - exercise training

KW - ion channel remodeling

KW - micro-RNAs

KW - sinoatrial node

KW - sinus bradycardia

U2 - 10.1161/CIRCRESAHA.117.311607

DO - 10.1161/CIRCRESAHA.117.311607

M3 - Journal article

C2 - 28821541

AN - SCOPUS:85031712012

SN - 0009-7322

VL - 121

SP - 1058

EP - 1068

JO - Circulation

JF - Circulation

IS - 9

ER -

Targeting miR-423-5p Reverses Exercise Training-Induced HCN4 Channel Remodeling and Sinus Bradycardia

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Keywords

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