ALPK3-deficient cardiomyocytes generated from patient-derived induced pluripotent stem cells and mutant human embryonic stem cells display abnormal calcium handling and establish that ALPK3 deficiency underlies familial cardiomyopathy

Dean G Phelan, David J Anderson, Sara E Howden, Raymond C B Wong, Peter F Hickey, Kate Pope, Gabrielle Rosalie Anne-Ma Wilson, Alice Pebay, Andrew M Davis, Steven Petrou, Andrew G Elefanty, Edouard G Stanley, Paul A James, Ivan Macciocca, Melanie Bahlo, Micheal M H Cheung, David J Amor, David A Elliott, Paul J Lockhart

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Abstract

AIMS: We identified a novel homozygous truncating mutation in the gene encoding alpha kinase 3 (ALPK3) in a family presenting with paediatric cardiomyopathy. A recent study identified biallelic truncating mutations of ALPK3 in three unrelated families; therefore, there is strong genetic evidence that ALPK3 mutation causes cardiomyopathy. This study aimed to clarify the mutation mechanism and investigate the molecular and cellular pathogenesis underlying ALPK3-mediated cardiomyopathy. METHODS AND RESULTS: We performed detailed clinical and genetic analyses of a consanguineous family, identifying a new ALPK3 mutation (c.3792G>A, p.W1264X) which undergoes nonsense-mediated decay in ex vivo and in vivo tissues. Ultra-structural analysis of cardiomyocytes derived from patient-specific and human ESC-derived stem cell lines lacking ALPK3 revealed disordered sarcomeres and intercalated discs. Multi-electrode array analysis and calcium imaging demonstrated an extended field potential duration and abnormal calcium handling in mutant contractile cultures. CONCLUSIONS: This study validates the genetic evidence, suggesting that mutations in ALPK3 can cause familial cardiomyopathy and demonstrates loss of function as the underlying genetic mechanism. We show that ALPK3-deficient cardiomyocytes derived from pluripotent stem cell models recapitulate the ultrastructural and electrophysiological defects observed in vivo. Analysis of differentiated contractile cultures identified abnormal calcium handling as a potential feature of cardiomyocytes lacking ALPK3, providing functional insights into the molecular mechanisms underlying ALPK3-mediated cardiomyopathy.
Original languageEnglish
Pages (from-to)1 - 5
Number of pages5
JournalEuropean Heart Journal
Volume37
Issue number33
DOIs
Publication statusPublished - 2016

Cite this

Phelan, Dean G ; Anderson, David J ; Howden, Sara E ; Wong, Raymond C B ; Hickey, Peter F ; Pope, Kate ; Wilson, Gabrielle Rosalie Anne-Ma ; Pebay, Alice ; Davis, Andrew M ; Petrou, Steven ; Elefanty, Andrew G ; Stanley, Edouard G ; James, Paul A ; Macciocca, Ivan ; Bahlo, Melanie ; Cheung, Micheal M H ; Amor, David J ; Elliott, David A ; Lockhart, Paul J. / ALPK3-deficient cardiomyocytes generated from patient-derived induced pluripotent stem cells and mutant human embryonic stem cells display abnormal calcium handling and establish that ALPK3 deficiency underlies familial cardiomyopathy. In: European Heart Journal. 2016 ; Vol. 37, No. 33. pp. 1 - 5.
@article{41788b3e3eb149509f6f92c516e60476,
title = "ALPK3-deficient cardiomyocytes generated from patient-derived induced pluripotent stem cells and mutant human embryonic stem cells display abnormal calcium handling and establish that ALPK3 deficiency underlies familial cardiomyopathy",
abstract = "AIMS: We identified a novel homozygous truncating mutation in the gene encoding alpha kinase 3 (ALPK3) in a family presenting with paediatric cardiomyopathy. A recent study identified biallelic truncating mutations of ALPK3 in three unrelated families; therefore, there is strong genetic evidence that ALPK3 mutation causes cardiomyopathy. This study aimed to clarify the mutation mechanism and investigate the molecular and cellular pathogenesis underlying ALPK3-mediated cardiomyopathy. METHODS AND RESULTS: We performed detailed clinical and genetic analyses of a consanguineous family, identifying a new ALPK3 mutation (c.3792G>A, p.W1264X) which undergoes nonsense-mediated decay in ex vivo and in vivo tissues. Ultra-structural analysis of cardiomyocytes derived from patient-specific and human ESC-derived stem cell lines lacking ALPK3 revealed disordered sarcomeres and intercalated discs. Multi-electrode array analysis and calcium imaging demonstrated an extended field potential duration and abnormal calcium handling in mutant contractile cultures. CONCLUSIONS: This study validates the genetic evidence, suggesting that mutations in ALPK3 can cause familial cardiomyopathy and demonstrates loss of function as the underlying genetic mechanism. We show that ALPK3-deficient cardiomyocytes derived from pluripotent stem cell models recapitulate the ultrastructural and electrophysiological defects observed in vivo. Analysis of differentiated contractile cultures identified abnormal calcium handling as a potential feature of cardiomyocytes lacking ALPK3, providing functional insights into the molecular mechanisms underlying ALPK3-mediated cardiomyopathy.",
author = "Phelan, {Dean G} and Anderson, {David J} and Howden, {Sara E} and Wong, {Raymond C B} and Hickey, {Peter F} and Kate Pope and Wilson, {Gabrielle Rosalie Anne-Ma} and Alice Pebay and Davis, {Andrew M} and Steven Petrou and Elefanty, {Andrew G} and Stanley, {Edouard G} and James, {Paul A} and Ivan Macciocca and Melanie Bahlo and Cheung, {Micheal M H} and Amor, {David J} and Elliott, {David A} and Lockhart, {Paul J}",
year = "2016",
doi = "10.1093/eurheartj/ehw160",
language = "English",
volume = "37",
pages = "1 -- 5",
journal = "European Heart Journal",
issn = "0195-668X",
publisher = "Oxford University Press",
number = "33",

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Phelan, DG, Anderson, DJ, Howden, SE, Wong, RCB, Hickey, PF, Pope, K, Wilson, GRA-M, Pebay, A, Davis, AM, Petrou, S, Elefanty, AG, Stanley, EG, James, PA, Macciocca, I, Bahlo, M, Cheung, MMH, Amor, DJ, Elliott, DA & Lockhart, PJ 2016, 'ALPK3-deficient cardiomyocytes generated from patient-derived induced pluripotent stem cells and mutant human embryonic stem cells display abnormal calcium handling and establish that ALPK3 deficiency underlies familial cardiomyopathy', European Heart Journal, vol. 37, no. 33, pp. 1 - 5. https://doi.org/10.1093/eurheartj/ehw160

ALPK3-deficient cardiomyocytes generated from patient-derived induced pluripotent stem cells and mutant human embryonic stem cells display abnormal calcium handling and establish that ALPK3 deficiency underlies familial cardiomyopathy. / Phelan, Dean G; Anderson, David J; Howden, Sara E; Wong, Raymond C B; Hickey, Peter F; Pope, Kate; Wilson, Gabrielle Rosalie Anne-Ma; Pebay, Alice; Davis, Andrew M; Petrou, Steven; Elefanty, Andrew G; Stanley, Edouard G; James, Paul A; Macciocca, Ivan; Bahlo, Melanie; Cheung, Micheal M H; Amor, David J; Elliott, David A; Lockhart, Paul J.

In: European Heart Journal, Vol. 37, No. 33, 2016, p. 1 - 5.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - ALPK3-deficient cardiomyocytes generated from patient-derived induced pluripotent stem cells and mutant human embryonic stem cells display abnormal calcium handling and establish that ALPK3 deficiency underlies familial cardiomyopathy

AU - Phelan, Dean G

AU - Anderson, David J

AU - Howden, Sara E

AU - Wong, Raymond C B

AU - Hickey, Peter F

AU - Pope, Kate

AU - Wilson, Gabrielle Rosalie Anne-Ma

AU - Pebay, Alice

AU - Davis, Andrew M

AU - Petrou, Steven

AU - Elefanty, Andrew G

AU - Stanley, Edouard G

AU - James, Paul A

AU - Macciocca, Ivan

AU - Bahlo, Melanie

AU - Cheung, Micheal M H

AU - Amor, David J

AU - Elliott, David A

AU - Lockhart, Paul J

PY - 2016

Y1 - 2016

N2 - AIMS: We identified a novel homozygous truncating mutation in the gene encoding alpha kinase 3 (ALPK3) in a family presenting with paediatric cardiomyopathy. A recent study identified biallelic truncating mutations of ALPK3 in three unrelated families; therefore, there is strong genetic evidence that ALPK3 mutation causes cardiomyopathy. This study aimed to clarify the mutation mechanism and investigate the molecular and cellular pathogenesis underlying ALPK3-mediated cardiomyopathy. METHODS AND RESULTS: We performed detailed clinical and genetic analyses of a consanguineous family, identifying a new ALPK3 mutation (c.3792G>A, p.W1264X) which undergoes nonsense-mediated decay in ex vivo and in vivo tissues. Ultra-structural analysis of cardiomyocytes derived from patient-specific and human ESC-derived stem cell lines lacking ALPK3 revealed disordered sarcomeres and intercalated discs. Multi-electrode array analysis and calcium imaging demonstrated an extended field potential duration and abnormal calcium handling in mutant contractile cultures. CONCLUSIONS: This study validates the genetic evidence, suggesting that mutations in ALPK3 can cause familial cardiomyopathy and demonstrates loss of function as the underlying genetic mechanism. We show that ALPK3-deficient cardiomyocytes derived from pluripotent stem cell models recapitulate the ultrastructural and electrophysiological defects observed in vivo. Analysis of differentiated contractile cultures identified abnormal calcium handling as a potential feature of cardiomyocytes lacking ALPK3, providing functional insights into the molecular mechanisms underlying ALPK3-mediated cardiomyopathy.

AB - AIMS: We identified a novel homozygous truncating mutation in the gene encoding alpha kinase 3 (ALPK3) in a family presenting with paediatric cardiomyopathy. A recent study identified biallelic truncating mutations of ALPK3 in three unrelated families; therefore, there is strong genetic evidence that ALPK3 mutation causes cardiomyopathy. This study aimed to clarify the mutation mechanism and investigate the molecular and cellular pathogenesis underlying ALPK3-mediated cardiomyopathy. METHODS AND RESULTS: We performed detailed clinical and genetic analyses of a consanguineous family, identifying a new ALPK3 mutation (c.3792G>A, p.W1264X) which undergoes nonsense-mediated decay in ex vivo and in vivo tissues. Ultra-structural analysis of cardiomyocytes derived from patient-specific and human ESC-derived stem cell lines lacking ALPK3 revealed disordered sarcomeres and intercalated discs. Multi-electrode array analysis and calcium imaging demonstrated an extended field potential duration and abnormal calcium handling in mutant contractile cultures. CONCLUSIONS: This study validates the genetic evidence, suggesting that mutations in ALPK3 can cause familial cardiomyopathy and demonstrates loss of function as the underlying genetic mechanism. We show that ALPK3-deficient cardiomyocytes derived from pluripotent stem cell models recapitulate the ultrastructural and electrophysiological defects observed in vivo. Analysis of differentiated contractile cultures identified abnormal calcium handling as a potential feature of cardiomyocytes lacking ALPK3, providing functional insights into the molecular mechanisms underlying ALPK3-mediated cardiomyopathy.

UR - http://www.ncbi.nlm.nih.gov/pubmed/27106955

U2 - 10.1093/eurheartj/ehw160

DO - 10.1093/eurheartj/ehw160

M3 - Article

VL - 37

SP - 1

EP - 5

JO - European Heart Journal

JF - European Heart Journal

SN - 0195-668X

IS - 33

ER -