PMID- 36257954
OWN - NLM
STAT- MEDLINE
DCOM- 20221020
LR  - 20230831
IS  - 2041-1723 (Electronic)
IS  - 2041-1723 (Linking)
VI  - 13
IP  - 1
DP  - 2022 Oct 18
TI  - Tmem65 is critical for the structure and function of the intercalated discs in 
      mouse hearts.
PG  - 6166
LID - 10.1038/s41467-022-33303-y [doi]
LID - 6166
AB  - The intercalated disc (ICD) is a unique membrane structure that is indispensable 
      to normal heart function, yet its structural organization is not completely 
      understood. Previously, we showed that the ICD-bound transmembrane protein 65 
      (Tmem65) was required for connexin43 (Cx43) localization and function in cultured 
      mouse neonatal cardiomyocytes. Here, we investigate the functional and cellular 
      effects of Tmem65 reductions on the myocardium in a mouse model by injecting CD1 
      mouse pups (3-7 days after birth) with recombinant adeno-associated virus 9 
      (rAAV9) harboring Tmem65 shRNA, which reduces Tmem65 expression by 90% in mouse 
      ventricles compared to scrambled shRNA injection. Tmem65 knockdown (KD) results 
      in increased mortality which is accompanied by eccentric hypertrophic 
      cardiomyopathy within 3 weeks of injection and progression to dilated 
      cardiomyopathy with severe cardiac fibrosis by 7 weeks post-injection. Tmem65 KD 
      hearts display depressed hemodynamics as measured echocardiographically as well 
      as slowed conduction in optical recording accompanied by prolonged PR intervals 
      and QRS duration in electrocardiograms. Immunoprecipitation and super-resolution 
      microscopy demonstrate a physical interaction between Tmem65 and sodium channel beta 
      subunit (beta1) in mouse hearts and this interaction appears to be required for both 
      the establishment of perinexal nanodomain structure and the localization of both 
      voltage-gated sodium channel 1.5 (NaV1.5) and Cx43 to ICDs. Despite the loss of 
      NaV1.5 at ICDs, whole-cell patch clamp electrophysiology did not reveal 
      reductions in Na(+) currents but did show reduced Ca(2+) and K(+) currents in 
      Tmem65 KD cardiomyocytes in comparison to control cells. We conclude that 
      disrupting Tmem65 function results in impaired ICD structure, abnormal cardiac 
      electrophysiology, and ultimately cardiomyopathy.
CI  - (c) 2022. The Author(s).
FAU - Teng, Allen C T
AU  - Teng ACT
AD  - Department of Physiology, Temerty Faculty of Medicine, University of Toronto, 
      Toronto, ON, M5S 1A8, Canada. Allen.Teng@utoronto.ca.
AD  - Translational Biology and Engineering Program, Ted Rogers Centre for Heart 
      Research, Toronto, ON, M5G 1M1, Canada. Allen.Teng@utoronto.ca.
FAU - Gu, Liyang
AU  - Gu L
AD  - Translational Biology and Engineering Program, Ted Rogers Centre for Heart 
      Research, Toronto, ON, M5G 1M1, Canada.
FAU - Di Paola, Michelle
AU  - Di Paola M
AD  - Department of Physiology, Temerty Faculty of Medicine, University of Toronto, 
      Toronto, ON, M5S 1A8, Canada.
AD  - Translational Biology and Engineering Program, Ted Rogers Centre for Heart 
      Research, Toronto, ON, M5G 1M1, Canada.
FAU - Lakin, Robert
AU  - Lakin R
AD  - Department of Biology, York University, Toronto, ON, M3J 1P3, Canada.
FAU - Williams, Zachary J
AU  - Williams ZJ
AD  - The Center for Heart and Reparative Medicine, Fralin Biomedical Research 
      Institute at Virginia Tech. Carilion, Roanoke, VA, 24016, USA.
AD  - Translational Biology Medicine and Health Graduate Program, Virginia Tech, 
      Roanoke, VA, 24016, USA.
FAU - Au, Aaron
AU  - Au A
AUID- ORCID: 0000-0001-5717-048X
AD  - Institute of Biomedical Engineering, Faculty of Applied Science and Engineering, 
      University of Toronto, Toronto, ON, M5S 3G9, Canada.
AD  - Donnelly Centre, University of Toronto, Toronto, ON, M5S 3E1, Canada.
FAU - Chen, Wenliang
AU  - Chen W
AD  - Department of Biology, York University, Toronto, ON, M3J 1P3, Canada.
FAU - Callaghan, Neal I
AU  - Callaghan NI
AUID- ORCID: 0000-0001-8214-3395
AD  - Translational Biology and Engineering Program, Ted Rogers Centre for Heart 
      Research, Toronto, ON, M5G 1M1, Canada.
AD  - Institute of Biomedical Engineering, Faculty of Applied Science and Engineering, 
      University of Toronto, Toronto, ON, M5S 3G9, Canada.
FAU - Zadeh, Farigol Hakem
AU  - Zadeh FH
AD  - Department of Physiology, Temerty Faculty of Medicine, University of Toronto, 
      Toronto, ON, M5S 1A8, Canada.
AD  - Translational Biology and Engineering Program, Ted Rogers Centre for Heart 
      Research, Toronto, ON, M5G 1M1, Canada.
FAU - Zhou, Yu-Qing
AU  - Zhou YQ
AD  - Translational Biology and Engineering Program, Ted Rogers Centre for Heart 
      Research, Toronto, ON, M5G 1M1, Canada.
AD  - Institute of Biomedical Engineering, Faculty of Applied Science and Engineering, 
      University of Toronto, Toronto, ON, M5S 3G9, Canada.
FAU - Fatah, Meena
AU  - Fatah M
AD  - The Labatt Family Heart Centre (Dept. of Pediatrics) and Translational Medicine, 
      The Hospital for Sick Children & Research Institute, University of Toronto, 
      Toronto, ON., M5G 1X8, Canada.
FAU - Chatterjee, Diptendu
AU  - Chatterjee D
AD  - The Labatt Family Heart Centre (Dept. of Pediatrics) and Translational Medicine, 
      The Hospital for Sick Children & Research Institute, University of Toronto, 
      Toronto, ON., M5G 1X8, Canada.
FAU - Jourdan, L Jane
AU  - Jourdan LJ
AD  - The Center for Heart and Reparative Medicine, Fralin Biomedical Research 
      Institute at Virginia Tech. Carilion, Roanoke, VA, 24016, USA.
AD  - Virginia Tech Carilion School of Medicine, Roanoke, VA, 24016, USA.
AD  - Department of Biomedical Engineering and Mechanics, Virginia Polytechnic 
      Institute and State University, Blacksburg, VA, 24060, USA.
FAU - Liu, Jack
AU  - Liu J
AD  - Department of Physiology, Temerty Faculty of Medicine, University of Toronto, 
      Toronto, ON, M5S 1A8, Canada.
FAU - Simmons, Craig A
AU  - Simmons CA
AUID- ORCID: 0000-0001-7729-1772
AD  - Translational Biology and Engineering Program, Ted Rogers Centre for Heart 
      Research, Toronto, ON, M5G 1M1, Canada.
AD  - Institute of Biomedical Engineering, Faculty of Applied Science and Engineering, 
      University of Toronto, Toronto, ON, M5S 3G9, Canada.
FAU - Kislinger, Thomas
AU  - Kislinger T
AUID- ORCID: 0000-0003-3525-5540
AD  - Princess Margaret Cancer Centre, University Health Network, Toronto, ON, M5G 1L7, 
      Canada.
AD  - Department of Medical Biophysics, University of Toronto, Toronto, ON, M5G 1L7, 
      Canada.
FAU - Yip, Christopher M
AU  - Yip CM
AUID- ORCID: 0000-0003-4507-556X
AD  - Institute of Biomedical Engineering, Faculty of Applied Science and Engineering, 
      University of Toronto, Toronto, ON, M5S 3G9, Canada.
AD  - Donnelly Centre, University of Toronto, Toronto, ON, M5S 3E1, Canada.
FAU - Backx, Peter H
AU  - Backx PH
AD  - Department of Biology, York University, Toronto, ON, M3J 1P3, Canada.
FAU - Gourdie, Robert G
AU  - Gourdie RG
AUID- ORCID: 0000-0001-6021-0796
AD  - The Center for Heart and Reparative Medicine, Fralin Biomedical Research 
      Institute at Virginia Tech. Carilion, Roanoke, VA, 24016, USA.
AD  - Virginia Tech Carilion School of Medicine, Roanoke, VA, 24016, USA.
AD  - Department of Biomedical Engineering and Mechanics, Virginia Polytechnic 
      Institute and State University, Blacksburg, VA, 24060, USA.
FAU - Hamilton, Robert M
AU  - Hamilton RM
AUID- ORCID: 0000-0002-5344-1278
AD  - The Labatt Family Heart Centre (Dept. of Pediatrics) and Translational Medicine, 
      The Hospital for Sick Children & Research Institute, University of Toronto, 
      Toronto, ON., M5G 1X8, Canada.
FAU - Gramolini, Anthony O
AU  - Gramolini AO
AUID- ORCID: 0000-0003-1109-2070
AD  - Department of Physiology, Temerty Faculty of Medicine, University of Toronto, 
      Toronto, ON, M5S 1A8, Canada. Anthony.Gramolini@utoronto.ca.
AD  - Translational Biology and Engineering Program, Ted Rogers Centre for Heart 
      Research, Toronto, ON, M5G 1M1, Canada. Anthony.Gramolini@utoronto.ca.
LA  - eng
GR  - R01 HL141855/HL/NHLBI NIH HHS/United States
GR  - R35 HL161237/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20221018
PL  - England
TA  - Nat Commun
JT  - Nature communications
JID - 101528555
RN  - 0 (Connexin 43)
RN  - 0 (NAV1.5 Voltage-Gated Sodium Channel)
RN  - 0 (RNA, Small Interfering)
RN  - 0 (Transcription Factors)
SB  - IM
MH  - Mice
MH  - Animals
MH  - *Connexin 43/genetics/metabolism
MH  - *NAV1.5 Voltage-Gated Sodium Channel/genetics/metabolism
MH  - RNA, Small Interfering/metabolism
MH  - Myocardium/metabolism
MH  - Myocytes, Cardiac/metabolism
MH  - Transcription Factors/metabolism
PMC - PMC9579145
COIS- The authors declare no competing interests.
EDAT- 2022/10/19 06:00
MHDA- 2022/10/21 06:00
PMCR- 2022/10/18
CRDT- 2022/10/18 23:15
PHST- 2020/06/08 00:00 [received]
PHST- 2022/09/07 00:00 [accepted]
PHST- 2022/10/18 23:15 [entrez]
PHST- 2022/10/19 06:00 [pubmed]
PHST- 2022/10/21 06:00 [medline]
PHST- 2022/10/18 00:00 [pmc-release]
AID - 10.1038/s41467-022-33303-y [pii]
AID - 33303 [pii]
AID - 10.1038/s41467-022-33303-y [doi]
PST - epublish
SO  - Nat Commun. 2022 Oct 18;13(1):6166. doi: 10.1038/s41467-022-33303-y.