Evaluation of dystrophin gene deletion patterns in a large Duchenne/Becker muscular dystrophy patient sample; 17 years experience from one Turkish Diagnostic Center

Öz

Duchenne/Becker muscular dystrophy (DMD/BMD) is an X-linked recessive disease results from mutations in the dystrophin gene. We established the deletion pattern profile in unrelated DMD/BMD patients using multiplex PCR (M-PCR). Methods: During 1998-2015, 1,385 unrelated DMD/BMD patients were admitted for genetic confirmation and/or exclusion of the disease. Deletion analysis in the dystrophin (DMD) gene was performed. Results: Of all patients admitted, 42.6 % deletion carriers (n=589) were detected, of which 180 (80.3 %) were carrying single exon deletions and 409 (14.8 %) multiple exon deletions. Deletions covering the major hotspot region were 80.3 %, the minor region 14.8% and 2.4% covered both regions. The mean age of diagnosis of patients with out-of-frame deletions (7.27 year) was notably lower than the cases with in frame deletions (17.54 year). No single exon 4 deletion was detected. Conclusions: When the known deletion hotspots are considered, the study population showed a similar deletion pattern with other populations. The mean age of patients with out-of-frame deletions were lower than mean age of those with in-frame deletions, in concordance with the reading frame hypothesis. Strikingly, no single exon 4 deletion was found, supporting the hypothesis that absence of it might have no functional consequences. 

Anahtar Kelimeler

• Dystrophin,Gene Deletion,Duchenne-Becker Muscular Dystrophy

Evaluation of dystrophin gene deletion patterns in a large Duchenne/Becker muscular dystrophy patient sample; 17 years experience from one Turkish Diagnostic Center

Öz

Background: Duchenne/Becker muscular dystrophy (DMD/BMD) is an X-linked recessive disease
results from mutations in the dystrophin gene. We established the deletion pattern profile in unrelated
DMD/BMD patients using multiplex PCR (M-PCR). Methods: During 1998-2015, 1,385 unrelated
Deletion
analysis in the dystrophin(DMD) gene was performed.Results: Of all patients admitted, 42.6% deletion
carriers (n=589) were detected, of which 180 (80.3 %)were carrying single exon deletions and 409 (14.8
%) multiple exon deletions. Deletions covering the major hotspot region were 80.3 %, the minor region
14.8% and 2.4% covered both regions. The mean age of diagnosis of patients with out-of-frame
deletions (7.27 year) was notably lower than the cases with in frame deletions (17.54 year). No single
exon 4 deletion was detected.Conclusions: When the known deletion hotspots are considered, the
study population showed a similar deletion pattern with other populations. The mean age of patients with
out-of-frame deletions were lower than mean age of those with in-frame deletions, in concordance with
the reading frame hypothesis. Strikingly, no single exon 4 deletion was found, supporting the hypothesis
that absence of it might have no functional consequences

Kaynakça

1. 10. White S, Kalf M, Liu Q, Villerius M, Engelsma D, Kriek M, Vollebregt E, Bakker B, van Ommen GJ, Breuning MH, den Dunnen JT. Comprehensive detection of genomic duplications and deletions in the DMD gene, by use of multiplex amplifiable probe hybridization. Am J Hum Genet 2002; 71: 365-374.
2. 11. Koenig M, Hoffman EP, Bertelson CJ, Monaco AP, Feener C, Kunkel LM. Complete cloning of the Duchenne muscular dystrophy (DMD) cDNA and preliminary genomic organization of the DMD gene in normal and affected individuals. Cell 1987; 50: 509-17.
3. 12. Nobile C, Marchi J, Nigro V, Roberts RG, Danieli GA. Exon-intron organization of the human dystrophin gene. Genomics 1997; 45: 421-424.
4. 13. Den-Dunnen JT, Grootscholten PM, Bakker E. Topography of the Duchenne muscular dystrophy 8d0d9 gene: FIGE and cDNA analysis of 194 cases reveals 115 deletions and 13 duplications. Am J Hum Genet 1999; 45: 835-847.
5. 14. Gillard EF, Chamberlain JS, Murphy EG, Duff CL, Smith B, Burghes AHM, Thompson MW, Sutherland J, Oss I, Bodrug SE, Klamut HJ, Ray PN, Worton RG. Molecular and phenotypic analysis of patients with deletions within the deletion-rich region of the Duchenne muscular dystrophy (DMD) gene. Am J Hum Genet 1989; 45: 507-520.
6. 15. Chamberlain JS, Gibbs RA, Ranier JE, Nguyen PN, Caskey CT. Deletion screening of the Duchenne muscular dystrophy locus via multiplex DNA amplification. Nucleic Acids Res 1988; 23: 11141-11156.
7. 16. Beggs AH, Koenig M, Boyce FM, Kunkel LM. Detection of 98 % of DMD/BMD gene deletions by polymerase chain reaction. Hum Genet 1990; 86: 45-48.
8. 17. Miller SA, Dykes DD, Polesky HF. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Research 1988; 16: 1215.

9. 18. DMD exonic deletions/duplications reading-frame checker 1.9. www.dmd.nl (Updated 2009)
10. 19. Hoffman EP, Kunkel LM. Dystrophin abnormalities in Duchenne/Becker muscular dystrophy. Neuron 1989; 2: 1019-1029.
11. 1. Davies KE, Pearson PL, Harper PS, Murray JM, O'Brien T, Sarfarazi M, Williamson R. Linkage analysis of two cloned DNA sequences flanking the Duchenne muscular dystrophy locus on the short arm of the human X chromosome. Nucl Acids Res 1983; 11: 2303-2312.
12. 20. Niemann-Seyde S, Slomski R, Rininsland F, Ellermeyer U, Kwiatkowska J, Reiss J. Molecular genetic analysis of 67 patients with Duchenne/Becker muscular dystrophy. Hum Genet 1992; 90: 65-70.
13. 21. Banerjee M, Verma IC. Are there ethnic differences in deletions in the dystrophin gene? Am J Med Genet 1997; 68: 152-157.
14. 22. Lisiecka D, Wigowska-Sowinska J, Kwiatkowska J, GalasZgorzalewicz, B, Slomski R. Molecular-genetic characteristics of mutations in dystrophin gene and clinical symptoms in Duchenne muscular dystrophy. Neurol Neurochir Pol 1998; 32: 1069-1079.
15. 23. Mutirangura A, Jongpiputvanich S, Norapucsunton T, Theamboonlers A, Srivuthana S, Promchainant C, Tumwasorn S, Sueblinvong T. Multiplex PCR to detect the dystrophin gene deletion in Thai patients. J Med Assoc Thai 1995; 78: 460-465.
16. 24. Yuge L, Hui L, Bingdi X. Detection of gene deletions in Chinese patients with Duchenne/Becker muscular dystrophy using cDNA probes and the polymerase chain reaction method. Life Sci 1999; 65: 863-869.
17. 25. Florentin L, Mavrou A, Kekou K, Metaxotou C. Deletion patterns of Duchenne and Becker muscular dystrophies in Greece. J Med Genet 1995; 32: 48-51.
18. 26. Coral-Vazquez R, Arenas D, Cisneros B, Peñaloza L, Salamanca F, Kofman S, Mercado R, Montañez C. Pattern of deletions of the dystrophin gene in MexicanDuchenne/Becker muscular dystrophy patients: the use of new designed primers for the analysis of the major deletion "hot spot" region. Am J Med Genet 1997;13: 240-246.
19. 27. Patiño A, Narbona J, García-Delgado M. Molecular analysis of the Duchennemuscular dystrophy gene in Spanish individuals: deletion detection and familialdiagnosis. Am J Med Genet 1995; 59: 182-187.
20. 28. Speer A, Kräft U, Hanke R, Grade K, Coutelle C, Wulff K, Wehnert M, Herrmann FH, Kadasi L, Kunert E, Müller U, Förster C, Wolf C, Szibor R. Deletion analysis of DMD/BMD families from the German Democratic Republic and selected regions of Czechoslovakia and Hungary. J Med Genet 1990; 27: 679-682.
21. 29. Baranzini SE, Giliberto F, Herrera M, Bernath V, Barreiro C, Garcia Erro M, Grippo J, Szijan I. Deletion patterns in Argentine patients with Duchenne andBecker muscular dystrophy. Neurol Res 1998; 20: 409-414.
22. 2. Kingston HM, Thomas NS, Pearson PL, Sarfarazi M, Harper PS. Genetic linkage between Becker muscular dystrophy and a polymorphic DNA sequence on the short arm of the X chromosome. J Med Genet 1983; 20: 255-258.
23. 30. Danieli GA, Mioni F, Müller CR, Vitiello L, Mostacciuolo ML, Grimm T. Patterns of deletions of the dystrophin gene in different European populations. Hum Genet1993; 91: 342-346.
24. 31. Lai PS, Takeshima Y, Adachi K, Van Tran K, Nguyen HT, Low PS, Matsuo M. Comparative study on deletions of the dystrophin gene in three Asian populations. J Hum Genet 2002; 47: 552-555.
25. 32. Gökgöz N, Kuseyri F, Topaloğlu H, Yüksel-Apak M, Kirdar B. Screening of deletions and RFLP analysis in Turkish DMD/BMD families by PCR. Clin Genet 1993; 43: 261-266.
26. 33. Dinçer P, Topaloğlu H, Ayter S, Ozgüç M, Taşdemir HA, Renda Y. Molecular deletion patterns in Turkish Duchenne and Becker muscular dystrophy patients. Brain Dev 1996; 18: 91-94.
27. 34. Önengüt S, Kavaslar GN, Battaloğlu E, Serdaroğlu P, Deymeer F, Ozdemir C, Calafell F, Tolun A. Deletion pattern in the dystrophin gene in Turks and a comparison with Europeans and Indians. Ann Hum Genet 2000; 64: 33-40.
28. 35. Ülgenalp A, Giray O, Bora E, Hizli T, Kurul S, Sağin-Saylam G, Karasoy H, Uran N, Dizdarer G, Tütüncüoğlu S, Dirik E, Ozkinay F, Erçal D. Deletion analysis and clinical correlations in patients with Xp21 linked muscular dystrophy. Turk J Pediatr 2004; 46: 333-338.
29. 36. Torelli S, Muntoni F. Alternative splicing of dystrophin exon 4 in normal human muscle. Hum Genet 1996; 97: 521-523.
30. 37. Cutiongco EM, Padilla CD, Takenaka K, Yamasaki Y, Matsuo M, Nishio H. More deletions in the 5' region than in the central region of the dystrophin gene were identified among Filipino Duchenne and Becker muscular dystrophy patients. Am J Med Genet 1995; 59: 266-267.
31. 38. Frisso G, Sampaolo S, Pastore L, Carlomagnoc A, Calisea RM, Di Ioriob G, Salvatore F. Novel deletion at the M and P promoters of the human dystrophin gene associated with a Duchenne muscular dystrophy. Neurmuscul Disord 2002; 12: 494-497.
32. 3. Hoffman EP, Brown RH Jr, Kunkel LM. Dystrophin: the protein product of the Duchenne muscular dystrophy locus. Cell 1987; 51: 919-928.
33. 4. Bushby KM, Thambyayah M, Gardner-Medwin D. Prevalence and incidence of Becker muscular dystrophy. Lancet 1991; 337: 1022-1024.
34. 5. Muntoni F, Torelli S, Ferlini A. Dystrophin and mutations: one gene, several proteins, multiple phenotypes. Lancet Neurol. 2003; 2: 731-740.
35. 6. Hoffman EP, Fischbeck KH, Brown RH, Johnson M, Medori R, Loire JD, Harris JB, Waterston R, Brooke M, Specht L, Kupsky W, Chamberlain J, Caskey CT, Shapiro F, Kunkel LM. Characterization of dystrophin in muscle-biopsy specimens from patients with Duchenne’s or Becker’s muscular dystrophy. N Engl J Med 1988; 318: 1363-1368.
36. 7. Monaco AP, Bertelson CJ, Liechti-Gallati S, Moser H, Kunkel LM. An explanation for the phenotypic differences between patients bearing partial deletions of the DMD locus. Genomics 1988; 2: 90-95.
37. 8. Forrest SM, Cross GS, Flint T, Speer A, Robson KJH, Davies KE. Further studies of gene deletions that cause Duchenne and Becker muscular dystrophies. Genomics 1988; 2: 109-114.
38. 9. Takeshima Y, Yagi M, Okizuka Y, Awano H, Zhang Z, Yamauchi Y, Nishio H, Matsuo M. Mutation spectrum of the dystrophin gene in 442 Duchenne/Becker muscular dystrophy cases from one Japanese referral center. J Hum Genet 2010; 55: 379-388.