Functional characterization of ABCC8 variants of unknown significance based on bioinformatics predictions, splicing assays, and protein analyses: Benefits for the accurate diagnosis of congenital hyperinsulinism.
ABCC8
KATP channel SUR1 subunit
congenital hyperinsulinism
in silico predictions
minigene splicing assays
variants of unknown significance
Journal
Human mutation
ISSN: 1098-1004
Titre abrégé: Hum Mutat
Pays: United States
ID NLM: 9215429
Informations de publication
Date de publication:
04 2021
04 2021
Historique:
revised:
06
12
2020
received:
12
08
2020
accepted:
31
12
2020
pubmed:
8
1
2021
medline:
1
4
2022
entrez:
7
1
2021
Statut:
ppublish
Résumé
ABCC8 encodes the SUR1 subunit of the β-cell ATP-sensitive potassium channel whose loss of function causes congenital hyperinsulinism (CHI). Molecular diagnosis is critical for optimal management of CHI patients. Unfortunately, assessing the impact of ABCC8 variants on RNA splicing remains very challenging as this gene is poorly expressed in leukocytes. Here, we performed bioinformatics analysis and cell-based minigene assays to assess the impact on splicing of 13 ABCC8 variants identified in 20 CHI patients. Next, channel properties of SUR1 proteins expected to originate from minigene-detected in-frame splicing defects were analyzed after ectopic expression in COSm6 cells. Out of the analyzed variants, seven induced out-of-frame splicing defects and were therefore classified as recessive pathogenic, whereas two led to skipping of in-frame exons. Channel functional analysis of the latter demonstrated their pathogenicity. Interestingly, the common rs757110 SNP increased exon skipping in our system suggesting that it may act as a disease modifier factor. Our strategy allowed determining the pathogenicity of all selected ABCC8 variants, and CHI-inheritance pattern for 16 out of the 20 patients. This study highlights the value of combining RNA and protein functional approaches in variant interpretation and reveals the minigene splicing assay as a new tool for CHI molecular diagnostics.
Identifiants
pubmed: 33410562
doi: 10.1002/humu.24164
pmc: PMC8049974
mid: NIHMS1661852
doi:
Substances chimiques
ABCC8 protein, human
0
Sulfonylurea Receptors
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
408-420Subventions
Organisme : NIDDK NIH HHS
ID : R01 DK057699
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK066485
Pays : United States
Informations de copyright
© 2021 Wiley Periodicals LLC.
Références
Eur J Nucl Med Mol Imaging. 2007 Dec;34(12):2120-8
pubmed: 17661030
J Appl Genet. 2019 May;60(2):231
pubmed: 30888641
PLoS Genet. 2016 Jan 13;12(1):e1005756
pubmed: 26761715
Genes (Basel). 2020 Apr 27;11(5):
pubmed: 32349249
Hum Mutat. 2020 Aug 2;:
pubmed: 32741062
Genome Res. 2018 Jan;28(1):100-110
pubmed: 29162642
Eur J Endocrinol. 2013 Mar 15;168(4):557-64
pubmed: 23345197
Genet Med. 2015 May;17(5):405-24
pubmed: 25741868
J Clin Endocrinol Metab. 2006 Mar;91(3):933-40
pubmed: 16403819
Hum Mutat. 2013 Nov;34(11):1547-57
pubmed: 23983145
Pediatr Radiol. 2012 Nov;42(11):1372-9
pubmed: 22885604
Cell. 2015 Oct 22;163(3):698-711
pubmed: 26496609
Hum Mutat. 2019 Nov;40(11):2044-2056
pubmed: 31237724
Hum Mutat. 2012 Aug;33(8):1228-38
pubmed: 22505045
Am J Hum Genet. 2013 Jan 10;92(1):131-6
pubmed: 23273570
Cell. 2017 Jan 12;168(1-2):101-110.e10
pubmed: 28086082
Hum Genet. 2017 Sep;136(9):1155-1172
pubmed: 28597072
Diabetes. 2011 Jun;60(6):1797-804
pubmed: 21536946
Am J Med Genet A. 2019 Nov;179(11):2214-2227
pubmed: 31464105
EMBO Rep. 2009 Aug;10(8):810-6
pubmed: 19648957
Oncogene. 2020 Jan;39(1):30-35
pubmed: 31467430
Physiol Rep. 2015 Sep;3(9):
pubmed: 26416970
Physiol Rev. 1998 Jan;78(1):227-45
pubmed: 9457174
Elife. 2017 Jan 16;6:
pubmed: 28092267
BMC Med. 2019 Jul 11;17(1):132
pubmed: 31291970
Hum Mutat. 2006 Feb;27(2):214
pubmed: 16429405
Hum Mutat. 2020 May;41(5):884-905
pubmed: 32027066
J Clin Endocrinol Metab. 2012 Jan;97(1):E94-9
pubmed: 22031516
Hum Mutat. 2020 Aug;41(8):1358-1364
pubmed: 32369867
Cancer Res. 2020 Apr 1;80(7):1374-1386
pubmed: 32046981
FEBS Lett. 2006 Aug 7;580(18):4449-56
pubmed: 16870183
Hepatology. 2009 Feb;49(2):553-67
pubmed: 19101985
J Pathol Clin Res. 2020 Jan;6(1):12-16
pubmed: 31577849
Cell. 2019 Jan 24;176(3):535-548.e24
pubmed: 30661751
Nucleic Acids Res. 2020 Feb 20;48(3):1600-1601
pubmed: 31863589
Mol Imaging Biol. 2013 Feb;15(1):97-105
pubmed: 22752652
Clin Endocrinol (Oxf). 2012 Feb;76(2):312-3
pubmed: 21851374
Am J Hum Genet. 2007 Mar;80(3):416-32
pubmed: 17273963
Science. 2015 Jan 9;347(6218):1254806
pubmed: 25525159
Diabetes. 2007 Sep;56(9):2339-48
pubmed: 17575084
J Med Genet. 2010 Nov;47(11):752-9
pubmed: 20685672
Nat Rev Genet. 2002 Apr;3(4):285-98
pubmed: 11967553
J Clin Endocrinol Metab. 2013 Feb;98(2):E355-63
pubmed: 23275527
FEBS Lett. 2005 Mar 28;579(9):1900-3
pubmed: 15792793
Genome Res. 2011 Aug;21(8):1360-74
pubmed: 21659425
Nucleic Acids Res. 2014;42(16):10681-97
pubmed: 25147205
Nat Rev Genet. 2007 Oct;8(10):749-61
pubmed: 17726481
Hum Genet. 2019 Jul;138(7):771-785
pubmed: 31168774
Diabetes Care. 2012 Feb;35(2):198-203
pubmed: 22190679
Nature. 2018 Oct;562(7726):217-222
pubmed: 30209399
Genet Med. 2020 Jun;22(6):1005-1014
pubmed: 32123317