Congenital hereditary endothelial dystrophy 1 (CHED1) and posterior polymorphous corneal dystrophy 1 (PPCD1) are autosomal-dominant corneal endothelial dystrophies that have been genetically mapped to overlapping loci on the short arm of chromosome 20. We combined genetic and genomic approaches to identify the cause of disease in extensive pedigrees comprising over 100 affected individuals. After exclusion of pathogenic coding, splice-site, and copy-number variations, a parallel approach using targeted and whole-genome sequencing facilitated the identification of pathogenic variants in a conserved region of the OVOL2 proximal promoter sequence in the index families (c.-339_361dup for CHED1 and c.-370T>C for PPCD1). Direct sequencing of the OVOL2 promoter in other unrelated affected individuals identified two additional mutations within the conserved proximal promoter sequence (c.-274T>G and c.-307T>C). OVOL2 encodes ovo-like zinc finger 2, a C2H2 zinc-finger transcription factor that regulates mesenchymal-to-epithelial transition and acts as a direct transcriptional repressor of the established PPCD-associated gene ZEB1. Interestingly, we did not detect OVOL2 expression in the normal corneal endothelium. Our in vitro data demonstrate that all four mutated OVOL2 promoters exhibited more transcriptional activity than the corresponding wild-type promoter, and we postulate that the mutations identified create cryptic cis-acting regulatory sequence binding sites that drive aberrant OVOL2 expression during endothelial cell development. Our data establish CHED1 and PPCD1 as allelic conditions and show that CHED1 represents the extreme of what can be considered a disease spectrum. They also implicate transcriptional dysregulation of OVOL2 as a common cause of dominantly inherited corneal endothelial dystrophies.

Department of Ophthalmology 1st Faculty of Medicine Charles University Prague and General University Hospital Prague U Nemocnice 2 Prague 128 08 Czech Republic

European Eye Clinic Lexum Antala Staška 1670 80 Prague 140 00 Czech Republic

Institute of Inherited Metabolic Disorders 1st Faculty of Medicine Charles University Prague and General University Hospital Prague Ke Karlovu 2 Prague 128 08 Czech Republic

Institute of Molecular Genetics Vídeňská 1083 Prague 142 20 Czech Republic

Moorfields Eye Hospital London EC1V 2PD UK

UCL Genetics Institute University College London London WC1E 6BT UK

UCL Institute of Ophthalmology University College London London EC1V 9EL UK

Weatherall Institute of Molecular Medicine University of Oxford John Radcliffe Hospital Oxford OX3 9DS UK

000      

00000naa a2200000 a 4500

001      

bmc16020040

003      

CZ-PrNML

005      

20201021104717.0

007      

ta

008      

160722s2016 xxu f 000 0|eng||

009      

AR

024    7_

$a 10.1016/j.ajhg.2015.11.018 $2 doi

024    7_

$a 10.1016/j.ajhg.2015.11.018 $2 doi

035    __

$a (PubMed)26749309

040    __

$a ABA008 $b cze $d ABA008 $e AACR2

041    0_

$a eng

044    __

$a xxu

100    1_

$a Davidson, Alice E $u UCL Institute of Ophthalmology, University College London, London EC1V 9EL, UK.

245    10

$a Autosomal-Dominant Corneal Endothelial Dystrophies CHED1 and PPCD1 Are Allelic Disorders Caused by Non-coding Mutations in the Promoter of OVOL2 / $c AE. Davidson, P. Liskova, CJ. Evans, L. Dudakova, L. Nosková, N. Pontikos, H. Hartmannová, K. Hodaňová, V. Stránecký, Z. Kozmík, HJ. Levis, N. Idigo, N. Sasai, GJ. Maher, J. Bellingham, N. Veli, ND. Ebenezer, ME. Cheetham, JT. Daniels, CM. Thaung, K. Jirsova, V. Plagnol, M. Filipec, S. Kmoch, SJ. Tuft, AJ. Hardcastle,

520    9_

$a Congenital hereditary endothelial dystrophy 1 (CHED1) and posterior polymorphous corneal dystrophy 1 (PPCD1) are autosomal-dominant corneal endothelial dystrophies that have been genetically mapped to overlapping loci on the short arm of chromosome 20. We combined genetic and genomic approaches to identify the cause of disease in extensive pedigrees comprising over 100 affected individuals. After exclusion of pathogenic coding, splice-site, and copy-number variations, a parallel approach using targeted and whole-genome sequencing facilitated the identification of pathogenic variants in a conserved region of the OVOL2 proximal promoter sequence in the index families (c.-339_361dup for CHED1 and c.-370T>C for PPCD1). Direct sequencing of the OVOL2 promoter in other unrelated affected individuals identified two additional mutations within the conserved proximal promoter sequence (c.-274T>G and c.-307T>C). OVOL2 encodes ovo-like zinc finger 2, a C2H2 zinc-finger transcription factor that regulates mesenchymal-to-epithelial transition and acts as a direct transcriptional repressor of the established PPCD-associated gene ZEB1. Interestingly, we did not detect OVOL2 expression in the normal corneal endothelium. Our in vitro data demonstrate that all four mutated OVOL2 promoters exhibited more transcriptional activity than the corresponding wild-type promoter, and we postulate that the mutations identified create cryptic cis-acting regulatory sequence binding sites that drive aberrant OVOL2 expression during endothelial cell development. Our data establish CHED1 and PPCD1 as allelic conditions and show that CHED1 represents the extreme of what can be considered a disease spectrum. They also implicate transcriptional dysregulation of OVOL2 as a common cause of dominantly inherited corneal endothelial dystrophies.

650    12

$a alely $7 D000483

650    _2

$a sekvence nukleotidů $7 D001483

650    _2

$a dědičné dystrofie rohovky $x genetika $7 D003317

650    _2

$a DNA $7 D004247

650    _2

$a ženské pohlaví $7 D005260

650    _2

$a lidé $7 D006801

650    _2

$a mužské pohlaví $7 D008297

650    12

$a mutace $7 D009154

650    _2

$a rodokmen $7 D010375

650    12

$a promotorové oblasti (genetika) $7 D011401

650    _2

$a sekvenční homologie nukleových kyselin $7 D012689

650    _2

$a transkripční faktory $x genetika $7 D014157

655    _2

$a časopisecké články $7 D016428

655    _2

$a práce podpořená grantem $7 D013485

700    1_

$a Lišková, Petra $u UCL Institute of Ophthalmology, University College London, London EC1V 9EL, UK; Institute of Inherited Metabolic Disorders, First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, Ke Karlovu 2, Prague 128 08, Czech Republic; Department of Ophthalmology, First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, U Nemocnice 2, Prague 128 08, Czech Republic. Electronic address: petra.liskova@lf1.cuni.cz. $7 xx0173361

700    1_

$a Evans, Cerys J $u UCL Institute of Ophthalmology, University College London, London EC1V 9EL, UK.

700    1_

$a Dudakova, Lubica $u Institute of Inherited Metabolic Disorders, First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, Ke Karlovu 2, Prague 128 08, Czech Republic.

700    1_

$a Nosková, Lenka $u Institute of Inherited Metabolic Disorders, First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, Ke Karlovu 2, Prague 128 08, Czech Republic. $7 xx0117961

700    1_

$a Pontikos, Nikolas $u UCL Institute of Ophthalmology, University College London, London EC1V 9EL, UK; UCL Genetics Institute, University College London, London WC1E 6BT, UK.

700    1_

$a Hartmannová, Hana $u Institute of Inherited Metabolic Disorders, First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, Ke Karlovu 2, Prague 128 08, Czech Republic. $7 xx0121900

700    1_

$a Hodaňová, Kateřina $u Institute of Inherited Metabolic Disorders, First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, Ke Karlovu 2, Prague 128 08, Czech Republic. $7 xx0074115

700    1_

$a Stránecký, Viktor $u Institute of Inherited Metabolic Disorders, First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, Ke Karlovu 2, Prague 128 08, Czech Republic. $7 xx0128943

700    1_

$a Kozmík, Zbyněk $u Institute of Molecular Genetics, Vídeňská 1083, Prague 142 20, Czech Republic.

700    1_

$a Levis, Hannah J $u UCL Institute of Ophthalmology, University College London, London EC1V 9EL, UK.

700    1_

$a Idigo, Nwamaka $u UCL Institute of Ophthalmology, University College London, London EC1V 9EL, UK.

700    1_

$a Sasai, Noriaki $u UCL Institute of Ophthalmology, University College London, London EC1V 9EL, UK.

700    1_

$a Maher, Geoffrey J $u Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DS, UK.

700    1_

$a Bellingham, James $u UCL Institute of Ophthalmology, University College London, London EC1V 9EL, UK.

700    1_

$a Veli, Neyme $u Moorfields Eye Hospital, London EC1V 2PD, UK.

700    1_

$a Ebenezer, Neil D $u UCL Institute of Ophthalmology, University College London, London EC1V 9EL, UK.

700    1_

$a Cheetham, Michael E $u UCL Institute of Ophthalmology, University College London, London EC1V 9EL, UK.

700    1_

$a Daniels, Julie T $u UCL Institute of Ophthalmology, University College London, London EC1V 9EL, UK.

700    1_

$a Thaung, Caroline M H $u UCL Institute of Ophthalmology, University College London, London EC1V 9EL, UK; Moorfields Eye Hospital, London EC1V 2PD, UK.

700    1_

$a Jirsová, Kateřina, $u Institute of Inherited Metabolic Disorders, First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, Ke Karlovu 2, Prague 128 08, Czech Republic. $d 1966- $7 xx0101780

700    1_

$a Plagnol, Vincent $u UCL Genetics Institute, University College London, London WC1E 6BT, UK.

700    1_

$a Filipec, Martin, $u European Eye Clinic Lexum, Antala Staška 1670/80, Prague 140 00, Czech Republic. $d 1955- $7 nlk20040147089

700    1_

$a Kmoch, Stanislav, $u Institute of Inherited Metabolic Disorders, First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, Ke Karlovu 2, Prague 128 08, Czech Republic. $d 1963- $7 xx0056529

700    1_

$a Tuft, Stephen J $u UCL Institute of Ophthalmology, University College London, London EC1V 9EL, UK; Moorfields Eye Hospital, London EC1V 2PD, UK.

700    1_

$a Hardcastle, Alison J $u UCL Institute of Ophthalmology, University College London, London EC1V 9EL, UK. Electronic address: a.hardcastle@ucl.ac.uk.

773    0_

$w MED00000254 $t American journal of human genetics $x 1537-6605 $g Roč. 98, č. 1 (2016), s. 75-89

856    41

$u https://pubmed.ncbi.nlm.nih.gov/26749309 $y Pubmed

910    __

$a ABA008 $b sig $c sign $y a $z 0

990    __

$a 20160722 $b ABA008

991    __

$a 20201021104713 $b ABA008

999    __

$a ok $b bmc $g 1154710 $s 944568

BAS    __

$a 3

BAS    __

$a PreBMC

BMC    __

$a 2016 $b 98 $c 1 $d 75-89 $e 20151231 $i 1537-6605 $m American journal of human genetics $n Am J Hum Genet $x MED00000254

GRA    __

$a NT13116 $p MZ0

GRA    __

$a NV15-28208A $p MZ0

LZP    __

$a Pubmed-20160722