Sequence variants with large effects on cardiac electrophysiology and disease

Norland, Kristjan; Sveinbjörnsson, Garðar; Þórólfsdóttir, Rósa B.; Davidsson, Olafur B.; Tragante, Vinicius; Rajamani, Sridharan; Helgadóttir, Anna; Grétarsdóttir, Sólveig; Setten, Jessica van; Asselbergs, Folkert W.; Sverrisson, Jon Th.; Stephensen, Sigurdur S; Óskarsson, Gylfi; Sigurðsson, Engilbert; Andersen, Karl; Daníelsen, Ragnar; Þorgeirsson, Guðmundur; Þorsteinsdóttir, Unnur; Arnar, Davíð O.; Sulem, Patrick; Hólm, Hilma; Guðbjartsson, Daníel F.; Stefánsson, Kāri

doi:10.1038/s41467-019-12682-9

Cited by 32 publications

(40 citation statements)

References 67 publications

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“…In four subsequent years, there has been just a single report that linked a missense sequence variation in the human ADPRHL1 locus to a specific clinical defect of the left ventricle through a genome-wide association study (GWAS) [23]. No other gene knockdown or knockout in other experimental model animal species has yet validated adprhl1 function during heart development.…”

Section: Introductionmentioning

confidence: 99%

Defective heart chamber growth and myofibrillogenesis after knockout of adprhl1 gene function by targeted disruption of the ancestral catalytic active site

et al. 2020

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ADP-ribosylhydrolase-like 1 (Adprhl1) is a pseudoenzyme expressed in the developing heart myocardium of all vertebrates. In the amphibian Xenopus laevis, knockdown of the two cardiac Adprhl1 protein species (40 and 23 kDa) causes failure of chamber outgrowth but this has only been demonstrated using antisense morpholinos that interfere with RNAsplicing. Transgenic production of 40 kDa Adprhl1 provides only part rescue of these defects. CRISPR/Cas9 technology now enables targeted mutation of the adprhl1 gene in G0-generation embryos with routine cleavage of all alleles. Testing multiple gRNAs distributed across the locus reveals exonic locations that encode critical amino acids for Adprhl1 function. The gRNA recording the highest frequency of a specific ventricle outgrowth phenotype directs Cas9 cleavage of an exon 6 sequence, where microhomology mediated endjoining biases subsequent DNA repairs towards three small in-frame deletions. Mutant alleles encode discrete loss of 1, 3 or 4 amino acids from a di-arginine (Arg271-Arg272) containing peptide loop at the centre of the ancestral ADP-ribosylhydrolase site. Thus despite lacking catalytic activity, it is the modified (adenosine-ribose) substrate binding cleft of Adprhl1 that fulfils an essential role during heart formation. Mutation results in striking loss of myofibril assembly in ventricle cardiomyocytes. The defects suggest Adprhl1 participation from the earliest stage of cardiac myofibrillogenesis and are consistent with previous MO results and Adprhl1 protein localization to actin filament Z-disc boundaries. A single nucleotide change to the gRNA sequence renders it inactive. Mice lacking Adprhl1 exons 3-4 are normal but production of the smaller ADPRHL1 species is unaffected, providing further evidence that cardiac activity is concentrated at the C-terminal protein portion.

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Section: Introductionmentioning

confidence: 99%

Defective heart chamber growth and myofibrillogenesis after knockout of adprhl1 gene function by targeted disruption of the ancestral catalytic active site

et al. 2020

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“…We identify several index variants in or near genes previously related to the cardiovascular system, either through disease associations or links to cardiovascular development and function. ALPK3, FHOD3, VCL and CAND2 has been implicated heart development and/or function 27,[30][31][32] , and TNFAIP2 is central in blood vessel formation 33 . Both ALPK3 and VCL are associated with cardiomyopathy 4,23,27 , and CAND2 is associated with atrial fibrillation 28 .…”

Section: Discussionmentioning

confidence: 99%

“…Most of the remaining index variants are in close proximity to genes previously associated with cardiovascular disease (ALPK3, VCL, CAND2, KLKB1, CYP4V2, HLA-DQB1) 4,23,[26][27][28][29] , cardiovascular development, structure and function (ALPK3, FHOD3, VCL, CAND2, TNFAIP2) 27,[30][31][32][33] , unspecific muscle injury (CAND2, ANO5) 20 , or processes such as coagulation/fibrinolysis, blood-pressure regulation and vascular inflammation (F12, KLKB1, HLA-DQB1, LMAN1) 29,34,35 . The F12/GRK6 locus has also been associated with chronic kidney disease 36 .…”

Section: Discovery Of Genetic Loci Associated With Ctnimentioning

confidence: 99%

Using human genetics to understand the causes and consequences of circulating cardiac troponin I in the general population

Moksnes¹,

Røsjø²,

Richmond³

et al. 2020

Preprint

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Circulating cardiac troponin proteins are associated with structural heart disease and predict incident cardiovascular disease in the general population. However, the genetic contribution to cardiac troponin I (cTnI) concentrations and its causal effect on cardiovascular phenotypes is unclear. We combine data from the Trondelag Health Study and the Generation Scotland Scottish Family Health Study and perform a genome-wide association study of high-sensitivity cTnI concentrations with 48 115 individuals. We identified 12 genetic loci (8 novel) associated with cTnI concentrations. Associated protein-altering variants highlighted putative functional genes: CAND2, HABP2, ANO5, APOH, FHOD3, TNFAIP2, KLKB1 and LMAN1. Using two-sample Mendelian randomization we confirmed the non-causal role of cTnI in acute myocardial infarction, but could not rule out a causal role for cTnI in heart failure. Using genetically informed methods for causal inference of cTnI helps inform the role and value of measuring cTnI in the general population.

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“…A recent human GWAS of the Icelander population has discovered a link between ADPRHL1 and heart ventricle function (Norland et al, 2019). They identified 190 sequence variations from a pool of 32.5 million Icelander SNPs and indels that associated with changes in a detailed analysis of electrocardiogram QRS parameters measured in 81 thousand individuals.…”

Section: Mapping Essential Regions Of the Adprhl1 Proteinmentioning

confidence: 99%

“…In four subsequent years, there has been just a single report that linked a missense sequence variation in the human ADPRHL1 locus to a specific clinical defect of the left ventricle through a genome-wide association study (GWAS) (Norland et al, 2019). No other gene knockdown or knockout in other experimental model animal species has yet validated adprhl1 function during heart development.…”

Section: Introductionmentioning

confidence: 99%

Defective heart chamber growth and myofibrillogenesis after knockout ofadprhl1gene function by targeted disruption of the ancestral catalytic active site

Smith

Towers

Demetriou

et al. 2020

Preprint

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1 Cardiac adprhl1 CRISPR knockout Smith et al. Mohun Highlights.Comparison of adprhl1 morpholinos. Knockdown of the two Xenopus cardiac Adprhl1 protein species (40 and 23 kDa) causes failure of ventricle outgrowth. CRISPR/Cas9 targeted gene mutation of adprhl1 with multiple gRNAs reveals exonic locations that encode critical amino acids for Adprhl1 function.Repair of DSBs at exon 6 yields small in-frame deletions that cause specific ventricle myofibril assembly defects.The deletions disturb a conserved di-arginine containing peptide loop at the centre of the ancestral substrate binding cleft/ADP-ribosylhydrolase site of this pseudoenzyme.Mice lacking Adprhl1 exons 3-4 are normal but production of the smaller ADPRHL1 species is unaffected, providing further evidence that cardiac activity is concentrated at the C-terminal protein portion. 2 Cardiac adprhl1 CRISPR knockout Smith et al. Mohun Abstract.ADP-ribosylhydrolase-like 1 (Adprhl1) is a pseudoenzyme expressed in the developing heart myocardium of all vertebrates. In the amphibian Xenopus laevis, knockdown of the two cardiac Adprhl1 protein species (40 and 23 kDa) causes failure of chamber outgrowth but this has only been demonstrated using antisense morpholinos that interfere with RNA-splicing. Transgenic production of 40 kDa Adprhl1 provides only part rescue of these defects. CRISPR/Cas9 technology now enables targeted mutation of the adprhl1 gene in G0-generation embryos with routine cleavage of all alleles. Testing multiple gRNAs distributed across the locus reveals exonic locations that encode critical amino acids for Adprhl1 function. The gRNA recording the highest frequency of a specific ventricle outgrowth phenotype directs Cas9 cleavage of an exon 6 sequence, where microhomology mediated end-joining biases subsequent DNA repairs towards three small in-frame deletions. Mutant alleles encode discrete loss of 1, 3 or 4 amino acids from a di-arginine (Arg271-Arg272) containing peptide loop at the centre of the ancestral ADP-ribosylhydrolase site. Thus despite lacking catalytic activity, it is the modified (adenosine-ribose) substrate binding cleft of Adprhl1 that fulfils an essential role during heart formation. Mutation results in striking loss of myofibril assembly in ventricle cardiomyocytes. The defects suggest Adprhl1 participation from the earliest stage of cardiac myofibrillogenesis and are consistent with previous MO results and Adprhl1 protein localization to actin filament Z-disc boundaries. A single nucleotide change to the gRNA sequence renders it inactive. Mice lacking Adprhl1 exons 3-4 are normal but production of the smaller ADPRHL1 species is unaffected, providing further evidence that cardiac activity is concentrated at the C-terminal protein portion.

show abstract

Sequence variants with large effects on cardiac electrophysiology and disease

Cited by 32 publications

References 67 publications

Defective heart chamber growth and myofibrillogenesis after knockout of adprhl1 gene function by targeted disruption of the ancestral catalytic active site

Defective heart chamber growth and myofibrillogenesis after knockout of adprhl1 gene function by targeted disruption of the ancestral catalytic active site

Using human genetics to understand the causes and consequences of circulating cardiac troponin I in the general population

Defective heart chamber growth and myofibrillogenesis after knockout ofadprhl1gene function by targeted disruption of the ancestral catalytic active site

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