Cardiac-targeted <italic style="font-style: italic">PIASy</italic> gene silencing mediates deSUMOylation of caveolin-3 and prevents ischemia/reperfusion-induced Na<sub>v</sub>1.5 downregulation and ventricular arrhythmias

Chen-Chen Hu; Xin Wei; Jin-Min Liu; Lin-Lin Han; Cheng-Kun Xia; Jing Wu; Tao You; A-Fang Zhu; Shang-Long Yao; Shi-Ying Yuan; Hao-Dong Xu; Zheng-Yuan Xia; Ting-Ting Wang; Wei-Ke Mao

doi:10.1186/s40779-022-00415-x

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Cardiac-targeted PIASy gene silencing mediates deSUMOylation of caveolin-3 and prevents ischemia/reperfusion-induced Na_v1.5 downregulation and ventricular arrhythmias

RESEARCH | Updated：2023-07-21

- Cardiac-targeted PIASy gene silencing mediates deSUMOylation of caveolin-3 and prevents ischemia/reperfusion-induced Na_v1.5 downregulation and ventricular arrhythmias
- Cardiac-targeted PIASy gene silencing mediates deSUMOylation of caveolin-3 and prevents ischemia/reperfusion-induced Na_v1.5 downregulation and ventricular arrhythmias
- MMR 2023年10卷第3期页码：342-358
- Affiliations：
  
  1.Department of Anesthesiology, Institute of Anesthesiology and Critical Care Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China
  2.Department of Cardiology, the Second Affiliated Hospital of Soochow University, Suzhou 215004, Jiangsu, China
  3.Department of Anesthesiology, Peking Union Medical College Hospital, CAMS and PUMC, Beijing 100730, China
  4.Department of Pathology, University of Washington, Seattle, WA 98195, USA
  5.State Key Laboratory of Pharmaceutical Biotechnology, the University of Hong Kong, Hong Kong 999077, China
  6.Department of Anesthesiology, Affiliated Hospital of Guangdong Medical University, Zhanjiang 524000, Guangdong, China
- Author bio：
  
  * wangtt201307@163.com;
  1984xH0556@hust.edu.cn
- Funds：
  
  This work was supported by grants from the National Natural Science Foundation of China(81770824;81270239)
- DOI：10.1186/s40779-022-00415-x
  中图分类号：
- 纸质出版：2023-06
- Accepted：
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Cardiac-targeted PIASy gene silencing mediates deSUMOylation of caveolin-3 and prevents ischemia/reperfusion-induced Na_v1.5 downregulation and ventricular arrhythmias[J]. MMR, 2023,10(3):342-358.

Cite this article as: Hu CC, Wei X, Liu JM, Han LL, Xia CK, Wu J, et al. Cardiac-targeted PIASy gene silencing mediates deSUMOylation of caveolin-3 and prevents ischemia/reperfusion-induced Na_v1.5 downregulation and ventricular arrhythmias. Mil Med Res. 2022;9(1):58.
Cardiac-targeted PIASy gene silencing mediates deSUMOylation of caveolin-3 and prevents ischemia/reperfusion-induced Na_v1.5 downregulation and ventricular arrhythmias[J]. MMR, 2023,10(3):342-358. DOI： 10.1186/s40779-022-00415-x.

Cite this article as: Hu CC, Wei X, Liu JM, Han LL, Xia CK, Wu J, et al. Cardiac-targeted PIASy gene silencing mediates deSUMOylation of caveolin-3 and prevents ischemia/reperfusion-induced Na_v1.5 downregulation and ventricular arrhythmias. Mil Med Res. 2022;9(1):58. DOI： 10.1186/s40779-022-00415-x.

摘要

Abstract

Background:

Abnormal myocardial voltage-gated sodium channel 1.5 (Na

1.5) expression and function cause lethal ventricular arrhythmias during myocardial ischemia–reperfusion (I/R). Protein inhibitor of activated STAT Y (PIASy)-mediated caveolin-3 (Cav-3) small ubiquitin-related modifier (SUMO) modification affects Cav-3 binding to the Na

1.5. PIASy activity is increased after myocardial I/R

but it is unclear whether this is attributable to plasma membrane Na

1.5 downregulation and ventricular arrhythmias.

Methods:

Using recombinant adeno-associated virus subtype 9 (AAV9)

rat cardiac PIASy was silenced using intraventricular injection of PIASy short hairpin RNA (shRNA). After two weeks

rat hearts were subjected to I/R and electrocardiography was performed to assess malignant arrhythmias. Tissues from peri-infarct areas of the left ventricle were collected for molecular biological measurements.

Results:

PIASy was upregulated by I/R (

0.01)

with increased SUMO2/3 modification of Cav-3 and reduced membrane Na

1.5 density (

0.01). AAV9-PIASy shRNA intraventricular injection into the rat heart down-regulated PIASy after I/R

at both mRNA and protein levels (

0.05

. Scramble-shRNA+I/R group)

decreased SUMO-modified Cav-3 levels

enhanced Cav-3 binding to Na

1.5

and prevented I/R-induced decrease of Na

1.5 and Cav-3 co-localization in the intercalated disc and lateral membrane. PIASy silencing in rat hearts reduced I/R-induced fatal arrhythmias

which was reflected by a modest decrease in the duration of ventricular fibrillation (VF;

0.05

. Scramble-shRNA+I/R group) and a significantly reduced arrhythmia score (

0.01

. Scramble-shRNA+I/R group). The anti-arrhythmic effects of PIASy silencing were also evidenced by decreased episodes of ventricular tachycardia (VT)

sustained VT and VF

especially at the time 5–10 min after ischemia (

0.05

. Scramble-shRNA+IR group). Using

in vitro

human embryonic kidney 293 T (HEK293T) cells and isolated adult rat cardiomyocyte models exposed to hypoxia/reoxygenation (H/R)

we confirmed that increased PIASy promoted Cav-3 modification by SUMO2/3 and Na

1.5/Cav-3 dissociation after H/R. Mutation of SUMO consensus lysine sites in Cav-3 (K38R or K144R) altered the membrane expression levels of Na

1.5 and Cav-3 before and after H/R in HEK293T cells.

Conclusions:

I/R-induced cardiac PIASy activation increased Cav-3 SUMOylation by SUMO2/3 and dysregulated Na

1.5-related ventricular arrhythmias. Cardiac-targeted PIASy silencing mediated Cav-3 deSUMOylation and partially prevented I/R-induced Na

1.5 downregulation in the plasma membrane of cardiomyocytes

and subsequent ventricular arrhythmias in rats. PIASy was identified as a potential therapeutic target for life-threatening arrhythmias in patients with ischemic heart diseases.

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references

Dong C , Wang Y , Ma A , Wang T . Life cycle of the cardiac voltage-gated sodium channel Na v 1.5 . Front Physiol . 2020 ; 11 : 609733 .

Nathan S , Gabelli SB , Yoder JB , Srinivasan L , Aldrich RW , Tomaselli GF , et al . Structural basis of cytoplasmic NaV1.5 and NaV1.4 regulation . J Gen Physiol . 2021 ; 153 ( 1 ): e202012722 .

El RM , Coles S , Musa H , Stevens TL , Wallace MJ , Murphy NP , et al . Altered expression of zonula occludens-1 affects cardiac Na + channels and increases susceptibility to ventricular arrhythmias . ells . 2022 ; 11 ( 4 ): 665 .

Zaklyazminskaya E , Dzemeshkevich S . The role of mutations in the SCN5A gene in cardiomyopathies . Biochim Biophys Acta . 2016 ; 1863 ( 7 Pt B ): 1799 - 805 .

Yang HQ , Perez-Hernandez M , Sanchez-Alonso J , Shevchuk A , Gorelik J , Rothenberg E , et al . Ankyrin-G mediates targeting of both Na + and KATP channels to the rat cardiac intercalated disc . Elife . 2020 ; 9 : e52373 .

Turan NN , Moshal KS , Roder K , Baggett BC , Kabakov AY , Dhakal S , et al . The endosomal trafficking regulator LITAF controls the cardiac Na v 1.5 channel via the ubiquitin ligase NEDD4–2 . J Biol Chem . 2020 ; 295 ( 52 ): 18148 - 59 .

Zhou J , Wang L , Zuo M , Wang X , Ahmed AS , Chen Q , et al . Cardiac sodium channel regulator MOG1 regulates cardiac morphogenesis and rhythm . Sci Rep . 2016 ; 6 : 21538 .

Matamoros M , Perez-Hernandez M , Guerrero-Serna G , Amoros I , Barana A , Nunez M , et al . Na v 1.5 N-terminal domain binding to alpha1-syntrophin increases membrane density of human Kir2.1, Kir2.2 and Na v 1.5 channels . Cardiovasc Res . 2016 ; 110 ( 2 ): 279 - 90 .

Eichel CA , Beuriot A , Chevalier MY , Rougier JS , Louault F , Dilanian G , et al . Lateral membrane-specific MAGUK CASK down-regulates Na v 1.5 channel in cardiac myocytes . Circ Res . 2016 ; 119 ( 4 ): 544 - 56 .

Balse E , Eichel C . The cardiac sodium channel and its protein partners . Handb Exp Pharmacol . 2018 ; 246 : 73 - 99 .

Schilling JM , Horikawa YT , Zemljic-Harpf AE , Vincent KP , Tyan L , Yu JK , et al . Electrophysiology and metabolism of caveolin-3-overexpressing mice . Basic Res Cardiol . 2016 ; 111 ( 3 ): 28 .

Dixon RE , Navedo MF , Binder MD , Santana LF . Mechanisms and physiological implications of cooperative gating of clustered ion channels . Physiol Rev . 2022 ; 102 ( 3 ): 1159 - 210 .

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 ; 61 : 102 - 10 .

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 ; 114 ( 20 ): 2104 - 12 .

Wilkinson KA , Henley JM . Mechanisms, regulation and consequences of protein SUMOylation . Biochem J . 2010 ; 428 ( 2 ): 133 - 45 .

Sahin U , de The H , Lallemand-Breitenbach V . Sumoylation in physiology, pathology and therapy . Cells . 2022 ; 11 ( 5 ): 814 .

Fuhs SR , Insel PA . Caveolin-3 undergoes SUMOylation by the SUMO E3 ligase PIASy: sumoylation affects G-protein-coupled receptor desensitization . J Biol Chem . 2011 ; 286 ( 17 ): 14830 - 41 .

Palvimo JJ . PIAS proteins as regulators of small ubiquitin-related modifier (SUMO) modifications and transcription . Biochem Soc Trans . 2007 ; 35 ( Pt 6 ): 1405 - 8 .

Yan Y , Ollila S , Wong I , Vallenius T , Palvimo JJ , Vaahtomeri K , et al . SUMOylation of AMPKα1 by PIAS4 specifically regulates mTORC1 signalling . Nat Commun . 2015 ; 6 : 8979 .

Diezko R , Suske G . Ligand binding reduces SUMOylation of the peroxisome proliferator-activated receptor gamma (PPARgamma) activation function 1 (AF1) domain . PLoS One . 2013 ; 8 ( 6 ): e66947 .

Wei X , Zhu A , Zhang Y , Yao S , Mao W . Pre- and delayed treatments with ranolazine ameliorate ventricular arrhythmias and Na v 1.5 downregulation in ischemic/reperfused rat hearts . J Cardiovasc Pharmacol . 2016 ; 68 ( 4 ): 269 - 79 .

Louch WE , Sheehan KA , Wolska BM . Methods in cardiomyocyte isolation, culture, and gene transfer . J Mol Cell Cardiol . 2011 ; 51 ( 3 ): 288 - 98 .

Piras BA , O'Connor DM , French BA . Systemic delivery of shRNA by AAV9 provides highly efficient knockdown of ubiquitously expressed GFP in mouse heart, but not liver . PLoS One . 2013 ; 8 ( 9 ): e75894 .

Bacchelli E , Loi E , Cameli C , Moi L , Vega-Benedetti AF , Blois S , et al . Analysis of a sardinian multiplex family with autism spectrum disorder points to post-synaptic density gene variants and identifies CAPG as a functionally relevant candidate gene . J Clin Med . 2019 ; 8 ( 2 ): 212 .

Wei M , Ma Y , Shen L , Xu Y , Liu L , Bu X , et al . NDRG2 regulates adherens junction integrity to restrict colitis and tumourigenesis . EBioMedicine . 2020 ; 61 : 103068 .

Curtis MJ , Walker MJ . Quantification of arrhythmias using scoring systems: an examination of seven scores in an in vivo model of regional myocardial ischaemia . Cardiovasc Res . 1988 ; 22 ( 9 ): 656 - 65 .

Kukkula A , Ojala VK , Mendez LM , Sistonen L , Elenius K , Sundvall M . Therapeutic potential of targeting the SUMO pathway in cancer . Cancers (Basel) . 2021 ; 13 ( 17 ): 4402 .

Chaanine AH , Nonnenmacher M , Kohlbrenner E , Jin D , Kovacic JC , Akar FG , et al . Effect of bortezomib on the efficacy of AAV9.SERCA2a treatment to preserve cardiac function in a rat pressure-overload model of heart failure . Gene Ther . 2014 ; 21 ( 4 ): 379 - 86 .

Ni L , Scott LJ , Campbell HM , Pan X , Alsina KM , Reynolds J , et al . Atrial-specific gene delivery using an adeno-associated viral vector . Circ Res . 2019 ; 124 ( 2 ): 256 - 62 .

Yang Y , Ma Z , Hu W , Wang D , Jiang S , Fan C , et al . Caveolin-1/-3: therapeutic targets for myocardial ischemia/reperfusion injury . Basic Res Cardiol . 2016 ; 111 ( 4 ): 45 .

Zhu A , Wei X , Zhang Y , You T , Yao S , Yuan S , et al . Propofol provides cardiac protection by suppressing the proteasome degradation of Caveolin-3 in ischemic/reperfused rat hearts . J Cardiovasc Pharmacol . 2017 ; 69 ( 3 ): 170 - 7 .

Vaidyanathan R , Reilly L , Eckhardt LL . Caveolin-3 microdomain: arrhythmia implications for potassium inward rectifier and cardiac sodium channel . Front Physiol . 2018 ; 9 : 1548 .

Marangoni S , Di Resta C , Rocchetti M , Barile L , Rizzetto R , Summa A , et al . A Brugada syndrome mutation (p.S216L) and its modulation by p.H558R polymorphism: standard and dynamic characterization . Cardiovasc Res . 2011 ; 91 ( 4 ): 606 - 16 .

Chang YC , Oram MK , Bielinsky AK . SUMO-targeted ubiquitin ligases and their functions in maintaining genome stability . Int J Mol Sci . 2021 ; 22 ( 10 ): 5391 .

Cai Q , Verma SC , Kumar P , Ma M , Robertson ES . Hypoxia inactivates the VHL tumor suppressor through PIASy-mediated SUMO modification . PLoS One . 2010 ; 5 ( 3 ): e9720 .

Mabb AM , Wuerzberger-Davis SM , Miyamoto S . PIASy mediates NEMO sumoylation and NF-kappaB activation in response to genotoxic stress . Nat Cell Biol . 2006 ; 8 ( 9 ): 986 - 93 .

Chen Y , Liu Z , Chen H , Huang X , Huang X , Lei Y , et al . p53 SUMOylation mediates aopp-induced endothelial senescence and apoptosis evasion . Front Cardiovasc Med . 2021 ; 8 : 795747 .

Han Y , Huang X , Cao X , Li Y , Gao L , Jia J , et al . SENP3-mediated TIP60 deSUMOylation is required for DNA-PKcs activity and DNA damage repair . MedComm . 2022 ; 3 ( 2 ): e123 .

Glynn P , Musa H , Wu X , Unudurthi SD , Little S , Qian L , et al . Voltage-gated sodium channel phosphorylation at ser571 regulates late current, arrhythmia, and cardiac function in vivo . Circulation . 2015 ; 132 ( 7 ): 567 - 77 .

Iqbal SM , Lemmens-Gruber R . Phosphorylation of cardiac voltage-gated sodium channel: Potential players with multiple dimensions . Acta Physiol (Oxf ) . 2019 ; 225 ( 3 ): e13210 .

Hallaq H , Wang DW , Kunic JD , George AJ , Wells KS , Murray KT . Activation of protein kinase C alters the intracellular distribution and mobility of cardiac Na + channels . Am J Physiol Heart Circ Physiol . 2012 ; 302 ( 3 ): H782 - 9 .

Zhao C , Wang L , Ma X , Zhu W , Yao L , Cui Y , et al . Cardiac Nav1.5 is modulated by ubiquitin protein ligase E3 component n-recognin UBR3 and 6 . J Cell Mol Med . 2015 ; 19 ( 9 ): 2143 - 52 .

Daimi H , Lozano-Velasco E , Aranega A , Franco D . Genomic and nongenomic regulatory mechanisms of the cardiac sodium channel in cardiac arrhythmias . Int J Mol Sci . 2022 ; 23 ( 3 ): 1381 .

Fouda MA , Ghovanloo MR , Ruben PC . Late sodium current: incomplete inactivation triggers seizures, myotonias, arrhythmias, and pain syndromes . J Physiol . 2022 ; 600 ( 12 ): 2835 - 51 .

Song Y , Shryock JC , Wagner S , Maier LS , Belardinelli L . Blocking late sodium current reduces hydrogen peroxide-induced arrhythmogenic activity and contractile dysfunction . J Pharmacol Exp Ther . 2006 ; 318 ( 1 ): 214 - 22 .

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Cardiac-targeted PIASy gene silencing mediates deSUMOylation of caveolin-3 and prevents ischemia/reperfusion-induced Nav1.5 downregulation and ventricular arrhythmias

Cardiac-targeted PIASy gene silencing mediates deSUMOylation of caveolin-3 and prevents ischemia/reperfusion-induced Nav1.5 downregulation and ventricular arrhythmias

DOI：10.1186/s40779-022-00415-x

摘要

Abstract

关键词

Keywords

references

Cardiac-targeted PIASy gene silencing mediates deSUMOylation of caveolin-3 and prevents ischemia/reperfusion-induced Na_v1.5 downregulation and ventricular arrhythmias

Cardiac-targeted PIASy gene silencing mediates deSUMOylation of caveolin-3 and prevents ischemia/reperfusion-induced Na_v1.5 downregulation and ventricular arrhythmias