TY - JOUR
T1 - A dominant mutation in RPE65 identified by whole-exome sequencing causes retinitis pigmentosa with choroidal involvement
AU - Bowne, Sara J.
AU - Humphries, Marian M.
AU - Sullivan, Lori S.
AU - Kenna, Paul F.
AU - Tam, Lawrence C.S.
AU - Kiang, Anna S.
AU - Campbell, Matthew
AU - Weinstock, George M.
AU - Koboldt, Daniel C.
AU - Ding, Li
AU - Fulton, Robert S.
AU - Sodergren, Erica J.
AU - Allman, Denis
AU - Millington-Ward, Sophia
AU - Palfi, Arpad
AU - McKee, Alex
AU - Blanton, Susan H.
AU - Slifer, Susan
AU - Konidari, Ioanna
AU - Farrar, G. Jane
AU - Daiger, Stephen P.
AU - Humphries, Peter
N1 - Funding Information:
We are grateful to TCD-G and TCD-H members who participated in this study, also Mr Hugh Nolan for clinical photography and Ms Hilary Dempsey for electroretinography. This work was supported by The Foundation Fighting Blindness, NIH EY007142, Fighting Blindness-Ireland, Science Foundation Ireland and Health Research Board of Ireland.
PY - 2011/10
Y1 - 2011/10
N2 - Linkage testing using Affymetrix 6.0 SNP Arrays mapped the disease locus in TCD-G, an Irish family with autosomal dominant retinitis pigmentosa (adRP), to an 8.8 Mb region on 1p31. Of 50 known genes in the region, 11 candidates, including RPE65 and PDE4B, were sequenced using di-deoxy capillary electrophoresis. Simultaneously, a subset of family members was analyzed using Agilent SureSelect All Exome capture, followed by sequencing on an Illumina GAIIx platform. Candidate gene and exome sequencing resulted in the identification of an Asp477Gly mutation in exon 13 of the RPE65 gene tracking with the disease in TCD-G. All coding exons of genes not sequenced to sufficient depth by next generation sequencing were sequenced by di-deoxy sequencing. No other potential disease-causing variants were found to segregate with disease in TCD-G. The Asp477Gly mutation was not present in Irish controls, but was found in a second Irish family provisionally diagnosed with choroideremia, bringing the combined maximum two-point LOD score to 5.3. Mutations in RPE65 are a known cause of recessive Leber congenital amaurosis (LCA) and recessive RP, but no dominant mutations have been reported. Protein modeling suggests that the Asp477Gly mutation may destabilize protein folding, and mutant RPE65 protein migrates marginally faster on SDS-PAGE, compared with wild type. Gene therapy for LCA patients with RPE65 mutations has shown great promise, raising the possibility of related therapies for dominant-acting mutations in this gene.
AB - Linkage testing using Affymetrix 6.0 SNP Arrays mapped the disease locus in TCD-G, an Irish family with autosomal dominant retinitis pigmentosa (adRP), to an 8.8 Mb region on 1p31. Of 50 known genes in the region, 11 candidates, including RPE65 and PDE4B, were sequenced using di-deoxy capillary electrophoresis. Simultaneously, a subset of family members was analyzed using Agilent SureSelect All Exome capture, followed by sequencing on an Illumina GAIIx platform. Candidate gene and exome sequencing resulted in the identification of an Asp477Gly mutation in exon 13 of the RPE65 gene tracking with the disease in TCD-G. All coding exons of genes not sequenced to sufficient depth by next generation sequencing were sequenced by di-deoxy sequencing. No other potential disease-causing variants were found to segregate with disease in TCD-G. The Asp477Gly mutation was not present in Irish controls, but was found in a second Irish family provisionally diagnosed with choroideremia, bringing the combined maximum two-point LOD score to 5.3. Mutations in RPE65 are a known cause of recessive Leber congenital amaurosis (LCA) and recessive RP, but no dominant mutations have been reported. Protein modeling suggests that the Asp477Gly mutation may destabilize protein folding, and mutant RPE65 protein migrates marginally faster on SDS-PAGE, compared with wild type. Gene therapy for LCA patients with RPE65 mutations has shown great promise, raising the possibility of related therapies for dominant-acting mutations in this gene.
KW - RPE65
KW - choroideremia
KW - exome capture
KW - next-generation sequencing
KW - retinitis pigmentosa
UR - http://www.scopus.com/inward/record.url?scp=80053050730&partnerID=8YFLogxK
U2 - 10.1038/ejhg.2011.86
DO - 10.1038/ejhg.2011.86
M3 - Article
C2 - 21654732
AN - SCOPUS:80053050730
SN - 1018-4813
VL - 19
SP - 1074
EP - 1081
JO - European Journal of Human Genetics
JF - European Journal of Human Genetics
IS - 10
ER -