TY - JOUR
T1 - Loss of the Atrial Fibrillation-Related Gene, Zfhx3, Results in Atrial Dilation and Arrhythmias
AU - Jameson, Heather S.
AU - Hanley, Alan
AU - Hill, Matthew C.
AU - Xiao, Ling
AU - Ye, Jiangchuan
AU - Bapat, Aneesh
AU - Ronzier, Elsa
AU - Hall, Amelia Weber
AU - Hucker, William J.
AU - Clauss, Sebastian
AU - Barazza, Miranda
AU - Silber, Elizabeth
AU - Mina, Julie A.
AU - Tucker, Nathan R.
AU - Mills, Robert W.
AU - Dong, Jin Tang
AU - Milan, David J.
AU - Ellinor, Patrick T.
N1 - Funding Information:
P.T. Ellinor is supported by grants from the National Institutes of Health (NIH; 1RO1HL092577, 1R01HL157635, 5R01HL139731), the American Heart Association Strategically Focused Research Networks (18SFRN34110082), and the European Union (Machine Learning Artificial Intelligence Early Detection Stroke Atrial Fibrillation (MAESTRIA) 965286). This research was also supported by the Carol and Roch Hillenbrand and the George L. Nardi, MD, funds at Massachusetts General Hospital. A. Hanley was supported by a postdoctoral fellowship from the Irish Cardiac Society; H.S. Jameson and W.J. Hucker were supported by NIH grant 5T32HL00720840. Dr Xiao was supported by an American Heart Association Career Development Award (20CDA35260081). N.R. Tucker is supported by the NIH Mentored Research Scientist Career Development Award (7K01HL140187). S. Clauss was supported by a Marie Curie International Outgoing Fellowship within the Seventh European Community Framework Program (PIOF-GA-2012-328352 to S. Clauss) and by the German Center for Cardiovascular Research (Deutsches Zentrum für Herz-Kreislauf-Forschung (DZHK); 81X2600210, 81X2600204).
Publisher Copyright:
© 2023 American Heart Association, Inc.
PY - 2023/8/4
Y1 - 2023/8/4
N2 - Background: ZFHX3 (zinc finger homeobox 3), a gene that encodes a large transcription factor, is at the second-most significantly associated locus with atrial fibrillation (AF), but its function in the heart is unknown. This study aims to identify causative genetic variation related to AF at the ZFHX3 locus and examine the impact of Zfhx3 loss on cardiac function in mice. Methods: CRISPR-Cas9 genome editing, chromatin immunoprecipitation, and luciferase assays in pluripotent stem cell-derived cardiomyocytes were used to identify causative genetic variation related to AF at the ZFHX3 locus. Cardiac function was assessed by echocardiography, magnetic resonance imaging, electrophysiology studies, calcium imaging, and RNA sequencing in mice with heterozygous and homozygous cardiomyocyte-restricted Zfhx3 loss (Zfhx3 Het and knockout, respectively). Human cardiac single-nucleus ATAC (assay for transposase-accessible chromatin)-sequencing data was analyzed to determine which genes in atrial cardiomyocytes are directly regulated by ZFHX3. Results: We found single-nucleotide polymorphism (SNP) rs12931021 modulates an enhancer regulating ZFHX3 expression, and the AF risk allele is associated with decreased ZFHX3 transcription. We observed a gene-dose response in AF susceptibility with Zfhx3 knockout mice having higher incidence, frequency, and burden of AF than Zfhx3 Het and wild-type mice, with alterations in conduction velocity, atrial action potential duration, calcium handling and the development of atrial enlargement and thrombus, and dilated cardiomyopathy. Zfhx3 loss results in atrial-specific differential effects on genes and signaling pathways involved in cardiac pathophysiology and AF. Conclusions: Our findings implicate ZFHX3 as the causative gene at the 16q22 locus for AF, and cardiac abnormalities caused by loss of cardiac Zfhx3 are due to atrial-specific dysregulation of pathways involved in AF susceptibility. Together, these data reveal a novel and important role for Zfhx3 in the control of cardiac genes and signaling pathways essential for normal atrial function.
AB - Background: ZFHX3 (zinc finger homeobox 3), a gene that encodes a large transcription factor, is at the second-most significantly associated locus with atrial fibrillation (AF), but its function in the heart is unknown. This study aims to identify causative genetic variation related to AF at the ZFHX3 locus and examine the impact of Zfhx3 loss on cardiac function in mice. Methods: CRISPR-Cas9 genome editing, chromatin immunoprecipitation, and luciferase assays in pluripotent stem cell-derived cardiomyocytes were used to identify causative genetic variation related to AF at the ZFHX3 locus. Cardiac function was assessed by echocardiography, magnetic resonance imaging, electrophysiology studies, calcium imaging, and RNA sequencing in mice with heterozygous and homozygous cardiomyocyte-restricted Zfhx3 loss (Zfhx3 Het and knockout, respectively). Human cardiac single-nucleus ATAC (assay for transposase-accessible chromatin)-sequencing data was analyzed to determine which genes in atrial cardiomyocytes are directly regulated by ZFHX3. Results: We found single-nucleotide polymorphism (SNP) rs12931021 modulates an enhancer regulating ZFHX3 expression, and the AF risk allele is associated with decreased ZFHX3 transcription. We observed a gene-dose response in AF susceptibility with Zfhx3 knockout mice having higher incidence, frequency, and burden of AF than Zfhx3 Het and wild-type mice, with alterations in conduction velocity, atrial action potential duration, calcium handling and the development of atrial enlargement and thrombus, and dilated cardiomyopathy. Zfhx3 loss results in atrial-specific differential effects on genes and signaling pathways involved in cardiac pathophysiology and AF. Conclusions: Our findings implicate ZFHX3 as the causative gene at the 16q22 locus for AF, and cardiac abnormalities caused by loss of cardiac Zfhx3 are due to atrial-specific dysregulation of pathways involved in AF susceptibility. Together, these data reveal a novel and important role for Zfhx3 in the control of cardiac genes and signaling pathways essential for normal atrial function.
KW - atrial fibrillation
KW - electrophysiology
KW - mice
KW - myocytes, cardiac
KW - transcription factors
UR - http://www.scopus.com/inward/record.url?scp=85166479095&partnerID=8YFLogxK
U2 - 10.1161/CIRCRESAHA.123.323029
DO - 10.1161/CIRCRESAHA.123.323029
M3 - Article
C2 - 37449401
AN - SCOPUS:85166479095
SN - 0009-7330
VL - 133
SP - 313
EP - 329
JO - Circulation research
JF - Circulation research
IS - 4
ER -