Visual Abstract

Introduction: Ventricular arrhythmia is the most important risk factor for sudden cardiac death (SCD) after acute myocardial infarction (MI) worldwide. However, the molecular mechanisms underlying these arrhythmias are complex and not completely understood. Objective: Here, we evaluated whether caveolin-3 (Cav3), the structural protein of caveolae, plays an important role in the therapeutic strategy for ventricular arrhythmias. Methods: A model of cardiac-specific overexpression of Cav3 was established to evaluate the incidence of ventricular arrhythmias after MI in mice. Ca2+ imaging was employed to detect the propensity of adult murine cardiomyocytes to generate arrhythmias, and immunoprecipitation and immunofluorescence were used to determine the relationship of proteins. Additionally, qRT-PCR and western blotting were used to detect the mRNA and protein expression. Results: We found that cardiac-specific overexpression of Cav3 delivered by a recombinant adeno-associated viral vector reduced the incidence of ventricular arrhythmias and SCD after MI in mice. Ca2+ imaging and western blotting revealed that overexpression of Cav3 reduced diastolic spontaneous Ca2+ waves by inhibiting the hyperphosphorylation of ryanodine receptor-2 (RyR2) at Ser2814, rather than at Ser2808, compared to in rAAV-red fluorescent protein control mice. Furthermore, we demonstrated that Cav3-regulated RYR2 hyperphosphorylation relied on plakophilin-2 in hypoxia-stimulated cultured cardiomyocytes by western blotting, immunoprecipitation, and immunofluorescence in vitro. Conclusions: Our results suggested a novel role for Cav3 in the prevention of ventricular arrhythmias, thereby identifying a new target for preventing SCD after MI.

1.
Zipes
DP
,
Camm
AJ
,
Borggrefe
M
,
Buxton
AE
,
Chaitman
B
,
Fromer
M
, et al;
American College of Cardiology/American Heart Association Task Force
;
European Society of Cardiology Committee for Practice Guidelines
;
European Heart Rhythm Association
;
Heart Rhythm Society
.
ACC/AHA/ESC 2006 guidelines for management of patients with ventricular arrhythmias and the prevention of sudden cardiac death: A report of the american college of Cardiology/American heart association task force and the european society of cardiology committee for practice guidelines (writing committee to develop guidelines for management of patients with ventricular arrhythmias and the prevention of sudden cardiac death): Developed in collaboration with the european heart rhythm association and the heart rhythm society
.
Circulation
.
2006
Sep
;
114
(
10
):
e385
484
.
[PubMed]
0009-7322
2.
Zipes
DP
,
Wellens
HJ
.
Sudden cardiac death
.
Circulation
.
1998
Nov
;
98
(
21
):
2334
51
.
[PubMed]
0009-7322
3.
Mitchell
LB
.
Treatment of ventricular arrhythmias after recovery from myocardial infarction
.
Annu Rev Med
.
1994
;
45
:
119
38
.
[PubMed]
0066-4219
4.
Xie
LH
,
Weiss
JN
.
Arrhythmogenic consequences of intracellular calcium waves
.
Am J Physiol Heart Circ Physiol
.
2009
Sep
;
297
(
3
):
H997
1002
.
[PubMed]
0363-6135
5.
Panneerselvam
M
,
Patel
HH
,
Roth
DM
.
Caveolins and heart diseases
.
Adv Exp Med Biol
.
2012
;
729
:
145
56
.
[PubMed]
0065-2598
6.
Rothberg
KG
,
Heuser
JE
,
Donzell
WC
,
Ying
YS
,
Glenney
JR
,
Anderson
RG
.
Caveolin, a protein component of caveolae membrane coats
.
Cell
.
1992
Feb
;
68
(
4
):
673
82
.
[PubMed]
0092-8674
7.
Song
KS
,
Scherer
PE
,
Tang
Z
,
Okamoto
T
,
Li
S
,
Chafel
M
, et al
Expression of caveolin-3 in skeletal, cardiac, and smooth muscle cells. Caveolin-3 is a component of the sarcolemma and co-fractionates with dystrophin and dystrophin-associated glycoproteins
.
J Biol Chem
.
1996
Jun
;
271
(
25
):
15160
5
.
[PubMed]
0021-9258
8.
Koga
A
,
Oka
N
,
Kikuchi
T
,
Miyazaki
H
,
Kato
S
,
Imaizumi
T
.
Adenovirus-mediated overexpression of caveolin-3 inhibits rat cardiomyocyte hypertrophy
.
Hypertension
.
2003
Aug
;
42
(
2
):
213
9
.
[PubMed]
0194-911X
9.
Tsutsumi
YM
,
Horikawa
YT
,
Jennings
MM
,
Kidd
MW
,
Niesman
IR
,
Yokoyama
U
, et al
Cardiac-specific overexpression of caveolin-3 induces endogenous cardiac protection by mimicking ischemic preconditioning
.
Circulation
.
2008
Nov
;
118
(
19
):
1979
88
.
[PubMed]
0009-7322
10.
Murfitt
L
,
Whiteley
G
,
Iqbal
MM
,
Kitmitto
A
.
Targeting caveolin-3 for the treatment of diabetic cardiomyopathy
.
Pharmacol Ther
.
2015
Jul
;
151
:
50
71
.
[PubMed]
0163-7258
11.
Hatzfeld
M
,
Wolf
A
,
Keil
R
.
Plakophilins in desmosomal adhesion and signaling
.
Cell Commun Adhes
.
2014
Feb
;
21
(
1
):
25
42
.
[PubMed]
1541-9061
12.
Novelli
V
,
Malkani
K
,
Cerrone
M
.
Pleiotropic phenotypes associated with PKP2 variants
.
Front Cardiovasc Med
.
2018
Dec
;
5
:
184
.
[PubMed]
2297-055X
13.
Cerrone
M
,
Montnach
J
,
Lin
X
,
Zhao
YT
,
Zhang
M
,
Agullo-Pascual
E
, et al
Plakophilin-2 is required for transcription of genes that control calcium cycling and cardiac rhythm
.
Nat Commun
.
2017
Jul
;
8
(
1
):
106
.
[PubMed]
2041-1723
14.
Xiao
X
,
Li
J
,
Samulski
RJ
.
Production of high-titer recombinant adeno-associated virus vectors in the absence of helper adenovirus
.
J Virol
.
1998
Mar
;
72
(
3
):
2224
32
.
[PubMed]
0022-538X
15.
Gao
E
,
Lei
YH
,
Shang
X
,
Huang
ZM
,
Zuo
L
,
Boucher
M
, et al
A novel and efficient model of coronary artery ligation and myocardial infarction in the mouse
.
Circ Res
.
2010
Dec
;
107
(
12
):
1445
53
.
[PubMed]
0009-7330
16.
Curtis
MJ
,
Hancox
JC
,
Farkas
A
,
Wainwright
CL
,
Stables
CL
,
Saint
DA
, et al
The Lambeth Conventions (II): guidelines for the study of animal and human ventricular and supraventricular arrhythmias
.
Pharmacol Ther
.
2013
Aug
;
139
(
2
):
213
48
.
[PubMed]
0163-7258
17.
Ma
SF
,
Luo
Y
,
Ding
YJ
,
Chen
Y
,
Pu
SX
,
Wu
HJ
, et al
Hydrogen sulfide targets the Cys320/Cys529 motif in kv4.2 to inhibit the ito potassium channels in cardiomyocytes and regularizes fatal arrhythmia in myocardial infarction
.
Antioxid Redox Signal
.
2015
Jul
;
23
(
2
):
129
47
.
[PubMed]
1523-0864
18.
de Almeida
PW
,
de Freitas Lima
R
,
de Morais Gomes
ER
,
Rocha-Resende
C
,
Roman-Campos
D
,
Gondim
AN
, et al
Functional cross-talk between aldosterone and angiotensin-(1-7) in ventricular myocytes
.
Hypertension
.
2013
Feb
;
61
(
2
):
425
30
.
[PubMed]
0194-911X
19.
Thireau
J
,
Karam
S
,
Fauconnier
J
,
Roberge
S
,
Cassan
C
,
Cazorla
O
, et al
Functional evidence for an active role of B-type natriuretic peptide in cardiac remodelling and pro-arrhythmogenicity
.
Cardiovasc Res
.
2012
Jul
;
95
(
1
):
59
68
.
[PubMed]
0008-6363
20.
Li
H
,
Huang
S
,
Wang
S
,
Wang
L
,
Qi
L
,
Zhang
Y
, et al
Relationship between annexin A7 and integrin β4 in autophagy
.
Int J Biochem Cell Biol
.
2013
Nov
;
45
(
11
):
2605
11
.
[PubMed]
1357-2725
21.
Huang
CL
.
Murine electrophysiological models of cardiac arrhythmogenesis
.
Physiol Rev
.
2017
Jan
;
97
(
1
):
283
409
.
[PubMed]
0031-9333
22.
Ter Keurs
HE
,
Boyden
PA
.
Calcium and arrhythmogenesis
.
Physiol Rev
.
2007
Apr
;
87
(
2
):
457
506
.
[PubMed]
0031-9333
23.
Belevych
AE
,
Terentyev
D
,
Terentyeva
R
,
Ho
HT
,
Gyorke
I
,
Bonilla
IM
, et al
Shortened Ca2+ signaling refractoriness underlies cellular arrhythmogenesis in a postinfarction model of sudden cardiac death
.
Circ Res
.
2012
Feb
;
110
(
4
):
569
77
.
[PubMed]
0009-7330
24.
Györke
S
,
Terentyev
D
.
Modulation of ryanodine receptor by luminal calcium and accessory proteins in health and cardiac disease
.
Cardiovasc Res
.
2008
Jan
;
77
(
2
):
245
55
.
[PubMed]
0008-6363
25.
Vatta
M
,
Ackerman
MJ
,
Ye
B
,
Makielski
JC
,
Ughanze
EE
,
Taylor
EW
, et al
Mutant caveolin-3 induces persistent late sodium current and is associated with long-QT syndrome
.
Circulation
.
2006
Nov
;
114
(
20
):
2104
12
.
[PubMed]
0009-7322
26.
Woodman
SE
,
Park
DS
,
Cohen
AW
,
Cheung
MW
,
Chandra
M
,
Shirani
J
, et al
Caveolin-3 knock-out mice develop a progressive cardiomyopathy and show hyperactivation of the p42/44 MAPK cascade
.
J Biol Chem
.
2002
Oct
;
277
(
41
):
38988
97
.
[PubMed]
0021-9258
27.
Horikawa
YT
,
Patel
HH
,
Tsutsumi
YM
,
Jennings
MM
,
Kidd
MW
,
Hagiwara
Y
, et al
Caveolin-3 expression and caveolae are required for isoflurane-induced cardiac protection from hypoxia and ischemia/reperfusion injury
.
J Mol Cell Cardiol
.
2008
Jan
;
44
(
1
):
123
30
.
[PubMed]
0022-2828
28.
Zhang
L
,
Yin
H
,
Jiao
L
,
Liu
T
,
Gao
Y
,
Shao
Y
, et al
Abnormal downregulation of caveolin-3 mediates the Pro-Fibrotic action of MicroRNA-22 in a model of myocardial infarction
.
Cell Physiol Biochem
.
2018
;
45
(
4
):
1641
53
.
[PubMed]
1015-8987
29.
Chen
X
,
Bonne
S
,
Hatzfeld
M
,
van Roy
F
,
Green
KJ
.
Protein binding and functional characterization of plakophilin 2. Evidence for its diverse roles in desmosomes and beta -catenin signaling
.
J Biol Chem
.
2002
Mar
;
277
(
12
):
10512
22
.
[PubMed]
0021-9258
30.
Laurita
KR
,
Rosenbaum
DS
.
Mechanisms and potential therapeutic targets for ventricular arrhythmias associated with impaired cardiac calcium cycling
.
J Mol Cell Cardiol
.
2008
Jan
;
44
(
1
):
31
43
.
[PubMed]
0022-2828
31.
Shannon
TR
,
Lew
WY
.
Diastolic release of calcium from the sarcoplasmic reticulum: a potential target for treating triggered arrhythmias and heart failure
.
J Am Coll Cardiol
.
2009
May
;
53
(
21
):
2006
8
.
[PubMed]
0735-1097
32.
Belevych
AE
,
Sansom
SE
,
Terentyeva
R
,
Ho
HT
,
Nishijima
Y
,
Martin
MM
, et al
MicroRNA-1 and -133 increase arrhythmogenesis in heart failure by dissociating phosphatase activity from RyR2 complex
.
PLoS One
.
2011
;
6
(
12
):
e28324
.
[PubMed]
1932-6203
33.
Ullrich
ND
,
Valdivia
HH
,
Niggli
E
.
PKA phosphorylation of cardiac ryanodine receptor modulates SR luminal Ca2+ sensitivity
.
J Mol Cell Cardiol
.
2012
Jul
;
53
(
1
):
33
42
.
[PubMed]
0022-2828
34.
Ather
S
,
Wang
W
,
Wang
Q
,
Li
N
,
Anderson
ME
,
Wehrens
XH
.
Inhibition of CaMKII phosphorylation of RyR2 prevents inducible ventricular arrhythmias in mice with Duchenne muscular dystrophy
.
Heart Rhythm
.
2013
Apr
;
10
(
4
):
592
9
.
[PubMed]
1547-5271
35.
Li
N
,
Wang
T
,
Wang
W
,
Cutler
MJ
,
Wang
Q
,
Voigt
N
, et al
Inhibition of CaMKII phosphorylation of RyR2 prevents induction of atrial fibrillation in FKBP12.6 knockout mice
.
Circ Res
.
2012
Feb
;
110
(
3
):
465
70
.
[PubMed]
0009-7330
36.
Cheng
J
,
Valdivia
CR
,
Vaidyanathan
R
,
Balijepalli
RC
,
Ackerman
MJ
,
Makielski
JC
.
Caveolin-3 suppresses late sodium current by inhibiting nNOS-dependent S-nitrosylation of SCN5A
.
J Mol Cell Cardiol
.
2013
Aug
;
61
:
102
10
.
[PubMed]
0022-2828
37.
Cerrone
M
,
Lin
X
,
Zhang
M
,
Agullo-Pascual
E
,
Pfenniger
A
,
Chkourko Gusky
H
, et al
Missense mutations in plakophilin-2 cause sodium current deficit and associate with a Brugada syndrome phenotype
.
Circulation
.
2014
Mar
;
129
(
10
):
1092
103
.
[PubMed]
0009-7322
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