New VMD2 gene mutations identified in patients affected by Best vitelliform macular dystrophy
- D Marchant1,
- K Yu2,
- K Bigot1,
- O Roche2,
- A Germain1,
- D Bonneau6,
- V Drouin-Garraud7,
- D F Schorderet3,
- F Munier8,
- D Schmidt2,
- P Le Neindre2,
- C Marsac1,
- M Menasche1,
- J L Dufier2,
- R Fischmeister5,
- C Hartzell4,
- M Abitbol1
- 1Centre de recherche thérapeutique en ophtalmologie, équipe d’accueil 2502 MENRT, Université René Descartes Paris V, Faculté de Médecine Necker-Enfants Malades, 156 rue de Vaugirard, Paris, France
- 2Département d’ophtalmologie, Centre Hospitalier Universitaire Necker-Enfants Malades, Paris, France
- 3IRO - Institut de Recherche en Ophtalmologie, Université de Lausanne et EPFL - Ecole polytechnique fédérale de Lausanne, Lausanne, Switzerland
- 4Department of Cell Biology, The Center for Neurodegenerative Disease, Emory University School of Medicine, Atlanta, Georgia, USA
- 5INSERM U769, Châtenay-Malabry, France
- 6Département de Génétique, Centre Hospitalier Universitaire d’Angers, Angers, France
- 7Département de Génétique, Centre Hospitalier Universitaire de Rouen, Rouen, France
- 8Service d’Ophtalmologie, Hôpital Jules Gonin, Lausanne, Switzerland
- Correspondence to: Dr M Abitbol Centre de recherche thérapeutique en ophtalmologie, Université René Descartes Paris V, Faculté de Médecine Necker-Enfants Malades, 156 rue de Vaugirard, 75015 Paris, France; abitbol{at}necker.fr Dr C Hartzell, Department of Cell Biology, Emory University School of Medicine, 615 Michael St, Atlanta, GA 30322, USA; criss.hartzell{at}emory.edu
- Received 5 June 2006
- Accepted 17 August 2006
- Revised 10 August 2006
- Published Online First 7 February 2007
Abstract
Purpose: The mutations responsible for Best vitelliform macular dystrophy (BVMD) are found in a gene called VMD2. The VMD2 gene encodes a transmembrane protein named bestrophin-1 (hBest1) which is a Ca2+-sensitive chloride channel. This study was performed to identify disease-specific mutations in 27 patients with BVMD. Because this disease is characterised by an alteration in Cl− channel function, patch clamp analysis was used to test the hypothesis that one of the VMD2 mutated variants causes the disease.
Methods: Direct sequencing analysis of the 11 VMD2 exons was performed to detect new abnormal sequences. The mutant of hBest1 was expressed in HEK-293 cells and the associated Cl− current was examined using whole-cell patch clamp analysis.
Results: Six new VMD2 mutations were identified, located exclusively in exons four, six and eight. One of these mutations (Q293H) was particularly severe. Patch clamp analysis of human embryonic kidney cells expressing the Q293H mutant showed that this mutant channel is non-functional. Furthermore, the Q293H mutant inhibited the function of wild-type bestrophin-1 channels in a dominant negative manner.
Conclusions: This study provides further support for the idea that mutations in VMD2 are a necessary factor for Best disease. However, because variable expressivity of VMD2 was observed in a family with the Q293H mutation, it is also clear that a disease-linked mutation in VMD2 is not sufficient to produce BVMD. The finding that the Q293H mutant does not form functional channels in the membrane could be explained either by disruption of channel conductance or gating mechanisms or by improper trafficking of the protein to the plasma membrane.
- AVMD, adult vitelliform macular dystrophy
- BVMD, Best vitelliform macular dystrophy
- EOG, electrooculogram
- HEK, human embryonic kidney
- mfERG, multifocal electroretinography
- OCT, optical coherence tomography
- RPE, retinal pigment epithelium
Footnotes
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Competing interests: None declared.
