TY - JOUR
T1 - Functional assessment of potential splice site variants in arrhythmogenic right ventricular dysplasia/cardiomyopathy
AU - Groeneweg, Judith A.
AU - Ummels, Amber
AU - Mulder, Marcel
AU - Bikker, Hennie
AU - van der Smagt, Jasper J.
AU - van Mil, Anneke M.
AU - Homfray, Tessa
AU - Post, Jan G.
AU - Elvan, Arif
AU - van der Heijden, Jeroen F.
AU - Houweling, Arjan C.
AU - Jongbloed, Jan D. H.
AU - Wilde, Arthur A. M.
AU - van Tintelen, J. Peter
AU - Hauer, Richard N.
AU - Dooijes, Dennis
PY - 2014
Y1 - 2014
N2 - Interpretation of genetic screening results in arrhythmogenic right ventricular dysplasia/cardiomyopathy (ARVD/C) often is difficult. Pathogenicity of variants with uncertain clinical significance may be predicted by software algorithms. However, functional assessment can unambiguously demonstrate the effect of such variants. The purpose of this study was to perform functional analysis of potential splice site variants in ARVD/C patients. Nine variants in desmosomal (PKP2, JUP, DSG2, DSC2) genes with potential RNA splicing effect were analyzed. The variants were found in patients who fulfilled 2010 ARVD/C Task Force Criteria (n = 7) or had suspected ARVD/C (n = 2). Total RNA was isolated from fresh blood samples and subjected to reverse transcriptase polymerase chain reaction. An effect on splicing was predicted by software algorithms for all variants. Of the 9 variants, 5 were intronic and 4 exonic. RNA analysis showed a functional effect on mRNA splicing by exon skipping, generation of new splice sites, or activation of cryptic sites in 6 variants. All 5 intronic variants tested severely impaired splicing. Only 1 of 4 exonic potential splice site variants was shown to have a deleterious effect on splicing. The remaining 3 exonic variants had no detectable effect on splicing, and heterozygous presence in mRNA confirmed biallelic expression. Six variants of uncertain clinical significance in the PKP2, JUP, and DSG2 genes showed a deleterious effect on mRNA splicing, indicating these are ARVD/C-related pathogenic splice site mutations. These results highlight the importance of functional assessment of potential splice site variants to improve patient care and facilitate cascade screening
AB - Interpretation of genetic screening results in arrhythmogenic right ventricular dysplasia/cardiomyopathy (ARVD/C) often is difficult. Pathogenicity of variants with uncertain clinical significance may be predicted by software algorithms. However, functional assessment can unambiguously demonstrate the effect of such variants. The purpose of this study was to perform functional analysis of potential splice site variants in ARVD/C patients. Nine variants in desmosomal (PKP2, JUP, DSG2, DSC2) genes with potential RNA splicing effect were analyzed. The variants were found in patients who fulfilled 2010 ARVD/C Task Force Criteria (n = 7) or had suspected ARVD/C (n = 2). Total RNA was isolated from fresh blood samples and subjected to reverse transcriptase polymerase chain reaction. An effect on splicing was predicted by software algorithms for all variants. Of the 9 variants, 5 were intronic and 4 exonic. RNA analysis showed a functional effect on mRNA splicing by exon skipping, generation of new splice sites, or activation of cryptic sites in 6 variants. All 5 intronic variants tested severely impaired splicing. Only 1 of 4 exonic potential splice site variants was shown to have a deleterious effect on splicing. The remaining 3 exonic variants had no detectable effect on splicing, and heterozygous presence in mRNA confirmed biallelic expression. Six variants of uncertain clinical significance in the PKP2, JUP, and DSG2 genes showed a deleterious effect on mRNA splicing, indicating these are ARVD/C-related pathogenic splice site mutations. These results highlight the importance of functional assessment of potential splice site variants to improve patient care and facilitate cascade screening
U2 - https://doi.org/10.1016/j.hrthm.2014.07.041
DO - https://doi.org/10.1016/j.hrthm.2014.07.041
M3 - Article
C2 - 25087486
SN - 1547-5271
VL - 11
SP - 2010
EP - 2017
JO - Heart Rhythm
JF - Heart Rhythm
IS - 11
ER -