Analysis of Mutations of Retinitis pigmentosa by Sequencing

Year 2017, Volume: 11 Issue: 1, 11 - 14, 30.04.2017

Nuray Altıntaş , Özge Sarıca Yılmaz Ali Soylu İlker Biçer

Abstract

In our study, we aimed to provide the first source of regional data obtained from molecular investigations by sequencing of selected exons of RPE65 and RPGR genes in Retinitis pigmentosa (RP) patients in Manisa province. Ethical committee approval was obtained and the study group consisted of 100 healthy volunteers with no clinical history of RP and RP with approved clinical manifestations in Ophthalmology Clinic of Manisa State Hospital. DNA samples were analyzed by Sequential Analysis and SeqScape software using ABI Prism 310 Genetic Analyzer and mutation tables. As a consequence of sequencing the sequence in the RPE65 gene in four patients with RP, two mutations known to be associated with the RPE65 gene have emerged in the BBS.E352E G> A Glu352Glu (Syn; 1056G> A) and R91Q G> A homozygote mutations were detected.Two RP patients had Bardet-Biedl syndrome.As a result of the sequence screening gene in RPGR , ORF15+1478 T>A heterozygote mutation in ORF15 exon of RPGR gene in 2 women patients , ORF15+1643 C>T and ORF15+1677 G>A polymorphisms in ORF15 exon of RPGR gene in 2 men patients were detected. Our results are originally given as regional first in Manisa and its region.

Keywords

RP, DNA Sequencing, ORF15, RPGR, RPE65

References

• [1] Jennifer D. Churchill, S. Bowne J, Sullivan S.L. Lewis A .R, Dianna K. Wheaton, Birch G.D, Branham K.E, Heckenlively J.R, Daiger S.P. Mutations in the X-Linked Retinitis Pigmentosa Genes RPGR and RP2 Found in 8.5% of Families with a Provisional Diagnosis of Autosomal Dominant Retinitis Pigmentosa.Investigative Ophthalmology & Visual Science, February 2013, Vol. 54, No. 2
• [2] Morımura H, Fıshman A.G, Grover S.A, Fulton A.B, Berson E.L,. Dryja T.P. Mutations in the RPE65 gene in patients with autosomal recessive retinitis pigmentosa or Leber congenital amaurosis .Proc. Natl. Acad. Sci. USA Vol. 95, pp. 3088–3093, March 1998. Medical Sciences .
• [3] Jiang,J, Wu X, Shen D,, Dong L, Jiao X, Hejtmancik J.F, Li N. Analysis of RP2 and RPGR Mutations in Five XLinked Chinese Families with Retinitis Pigmentosa. Scientific Reports | 7:44465 | DOI: 10.1038/srep44465 .Published: 15 March 2017.
• [4] Wang Y, Guo1 L, Cai S.P, Dai M, Yang O, Yu1, Yan W.N, Zhou X, Fu J, Guo X, Han P, Wang J, Liu X .Exome Sequencing Identifies Compound Heterozygous Mutations in CYP4V2 in a Pedigree with Retinitis Pigmentosa.PLoS ONE | PLoS ONE.| May 2012 | Volume 7 | Issue 5 | e33673.
• [5] Yoon C.K, Kim N.K.D, Joung J.G , Shin J.Y, Park J.H., Eum H.H, Lee H, Park W.Y, Yu H.G. The diagnostic application of targeted re-sequencing in Korean patients with retinitis pigmentosa.Yoon et al. BMC Genomics (2015) 16:515 DOI 10.1186/s12864-015-1723-x
• [6] Jacobson SG, Cideciyan AV, Aleman TS, Sumaroka A, Schwartz SB, Windsor EAM, Roman AJ, Heon E, Stone EM and Thompson DA. RDH12 and RPE65, Visual Cycle Genes Causing Leber Congenital Amaurosis, Differ in Disease Expression. Investigate Ophtalmology & Visual Science. January 2007, Vol. 48, No. 1: 332-338.
• [7] Philp AR, Jin M, Li S, Schindler EI, Iannaccone A, Lam BL, Weleber RG, Fishman GA, Jacobson SG, Mullins RF, Travis GH, Stone EM. Predicting the Pathogenicity of RPE65 Mutations. Human Mutation. August 2009; 30(8): 1183-1188.
• [8] Thompson AD, Gyürüs P, Fleiscer LL, Bingham EL, McHenry CL,Apfelstedt-Sylla E,Zrenner E,Lorenz B,Richards JE,Jacobson SG,Sieving PA and Gal A. Genetics and Phenotypes of RPE65 Mutations in Inherited Retinal Degeneration. Investigative Ophthalmology & Visual Science, December 2000, Vol. 41, No. 13: 4293-4299.
• [9] Simovich MJ, Miller B, Ezeldin H, Kirkland BT, McLeod G, Fulmer C, Nathans J, Jacobson SG and Pittler SJ. Four Novel Mutations in the RPE65 Gene in Patients With Leber Congenital Amaurosis.Human Mutation. August 2001, Volume 18, Issue 2, page 164-168.
• [10] Matri LE, Ambresin A, Schorderet DF, Kawasaki A,Seeliger MW, Wenzel A, Arsenijevic Y, Borruat FX, Munier FL. Phenotype of three consanguineous Tunisian families with early-onset retinal degeneration caused by an R91W homozygous mutation in the RPE65 gene. Clinical Investigation. 2006; 244: 1104-1112.
• [11] Jacobson SG, Cideciyan AV, Aleman TS, Sumaroka A, Windsor EAM, Schwartz SB, Heon E and Stone EM. Photoreceptor Layer Topography in Children with Leber Congenital Amaurosis Caused by RPE65 Mutations. Investigative Ophthalmology & Visual Science. October 2008, Vol. 49, No. 10: 4573-4577.
• [12] Mamatha G, Srilekha S, Meenakshi S, Kumaramanickavel G. Screening of the RPE65 Gene in the Asian Indian Patients with Leber Congenital Amaurosis. Ophthalmic Genetics, 2008, 29: 73-78.
• [13] Pusch CM, Broghammer M, Jurklies B, Besch D, Jacobi FK. Ten novel ORF15 mutations confirm mutational hot spot in the RPGR gene in European patients with Xlinked retinitis pigmentosa. Hum Mutat. 2002; 20(5):405.
• [14] Kellner U, Kellner S, Weber BH, Fiebig B, Weinitz S, Ruether K. Lipofuscinand melanin-related fundus autofluorescence visualize different retinal pigment epithelial alterations in patients with retinitis pigmentosa. Eye (Lond). 2009 Jun;23(6):1349-59.