Genetic characterization in symptomatic female DMD carriers: lack of relationship between X-inactivation, transcriptional DMD allele balancing and phenotype

BMC Medical Genetics

Alessandra Ferlini³  Tiziana Mongini⁸  Luciano Merlini³  Enrico Bertini²  Eugenio Mercuri⁹  Domenico De Grandis⁵  Liliana Vercelli⁸  Antonella Pini⁷  Silvana Tedeschi⁶  Gabriele Siciliano⁴  Giulia Ricci⁴  Marika Pane⁹  Adele D’Amico²  Rita Selvatici³  Patrizia Sabatelli¹  Maria S Falzarano³  Matteo Bovolenta³  Annarita Armaroli³  Chiara Scotton³  Francesca Gualandi³  Simona Brioschi³ 

[1] Istituto Di Genetica Molecolare-CNR, Istituto Ortopedico Rizzoli, Bologna, Italy;Department of Laboratory Medicine, Unit of Molecular Medicine, Bambino Gesù Hospital, Rome, Italy;Section of Medical Genetics, Department of Experimental and Diagnostic Medicine, University of Ferrara, Ferrara, Italy;Department of Neuroscience, Neurological Clinic, University of Pisa, Pisa, Italy;Division of Neurology, Department of Neuroscience, Civile Hospital Santa Maria della Misericordia, Rovigo, Italy;Laboratory of Medical Genetics, I.R.C.C.S. Foundation Ca’ Granda, Maggiore Hospital, Policlinico, Milan, Italy;Child Neurology and Psychiatry Unit, Maggiore Hospital, Bologna, Italy;Neuromuscular Center, S.G. Battista Hospital, University of Turin, Turin, Italy;Department of Child Neurology and Psychiatry, Catholic University, Rome, Italy

关键词: Transcriptional balancing;    X-inactivation;    Female carriers;    Dystrophinopathy;   

Others  :  1177812
DOI  :  10.1186/1471-2350-13-73

 received in 2012-04-11, accepted in 2012-08-08,  发布年份 2012

【 摘 要 】

Background

Although Duchenne and Becker muscular dystrophies, X-linked recessive myopathies, predominantly affect males, a clinically significant proportion of females manifesting symptoms have also been reported. They represent an heterogeneous group characterized by variable degrees of muscle weakness and/or cardiac involvement. Though preferential inactivation of the normal X chromosome has long been considered the principal mechanism behind disease manifestation in these females, supporting evidence is controversial.

Methods

Eighteen females showing a mosaic pattern of dystrophin expression on muscle biopsy were recruited and classified as symptomatic (7) or asymptomatic (11), based on the presence or absence of muscle weakness. The causative DMD gene mutations were identified in all cases, and the X-inactivation pattern was assessed in muscle DNA. Transcriptional analysis in muscles was performed in all females, and relative quantification of wild-type and mutated transcripts was also performed in 9 carriers. Dystrophin protein was quantified by immunoblotting in 2 females.

Results

The study highlighted a lack of relationship between dystrophic phenotype and X-inactivation pattern in females; skewed X-inactivation was found in 2 out of 6 symptomatic carriers and in 5 out of 11 asymptomatic carriers. All females were characterized by biallelic transcription, but no association was found between X-inactivation pattern and allele transcriptional balancing. Either a prevalence of wild-type transcript or equal proportions of wild-type and mutated RNAs was observed in both symptomatic and asymptomatic females. Moreover, very similar levels of total and wild-type transcripts were identified in the two groups of carriers.

Conclusions

This is the first study deeply exploring the DMD transcriptional behaviour in a cohort of female carriers. Notably, no relationship between X-inactivation pattern and transcriptional behaviour of DMD gene was observed, suggesting that the two mechanisms are regulated independently. Moreover, neither the total DMD transcript level, nor the relative proportion of the wild-type transcript do correlate with the symptomatic phenotype.

【 授权许可】

   
2012 Brioschi et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

【 参考文献 】

• [1]Moser H, Emery AE: The manifesting carrier in Duchenne muscular dystrophy. Clin Genet 1974, 5(4):271-284.
• [2]Norman A, Harper P: A survey of manifesting carriers of Duchenne and Becker muscular dystrophy in Wales. Clin Genet 1989, 36(1):31-37.
• [3]Comi LI, Nigro G, Politano L, Petretta VR: The cardiomyopathy of Duchenne/Becker consultands. Int J Cardiol 1992, 34(3):297-305.
• [4]Kinoshita H, Goto Y, Ishikawa M, Uemura T, Matsumoto K, Hayashi YK, Arahata K, Nonaka I: A carrier of Duchenne muscular dystrophy with dilated cardiomyopathy but no skeletal muscle symptom. Brain Dev 1995, 17(3):202-205.
• [5]Hoogerwaard EM, Bakker E, Ippel PF, Oosterwijk JC, Majoor-Krakauer DF, Leschot NJ, Van Essen AJ, Brunner HG, van der Wouw PA, Wilde AA, de Visser M: Signs and symptoms of Duchenne muscular dystrophy and Becker muscular dystrophy among carriers in the Netherlands: a cohort study. Lancet 1999, 353:2116-2119.
• [6]Chelly J, Marlhens F, Le Marec B, Jeanpierre M, Lambert M, Hamard G, Dutrillaux B, Kaplan JC: De novo DNA microdeletion in a girl with Turner syndrome and Duchenne muscular dystrophy. Hum Genet 1986, 74(2):193-196.
• [7]Quan F, Janas J, Toth-Fejel S, Johnson DB, Wolford JK, Popovich BW: Uniparental disomy of the entire X chromosome in a female with Duchenne muscular dystrophy. Am J Hum Genet 1997, 60(1):160-165.
• [8]Puck JM, Willard HF: X inactivation in females with X-linked disease. N Engl J Med 1998, 338(5):325-8.9.
• [9]Brown CJ, Robinson WP: The causes and consequences of random and non-random X chromosome inactivation in humans. Clin Genet 2000, 58(5):353-6.10.
• [10]Boyd Y, Buckle V, Holt S, Munro E, Hunter D, Craig I: Muscular dystrophy in girls with X;autosome translocations. J Med Genet 1986, 23(6):484-490.
• [11]Wenger SL, Steele MW, Hoffman EP, Barmada MA, Wessel HB: X inactivation and dystrophin studies in a t(X;12) female: evidence for biochemical normalization in Duchenne muscular dystrophy carriers. Am J Med Genet 1992, 43(6):1012-1015.
• [12]Pena SD, Karpati G, Carpenter S, Fraser FC: The clinical consequences of X-chromosome inactivation: Duchenne muscular dystrophy in one of monozygotic twins. J Neurol Sci 1987, 79(3):337-344.
• [13]Bonilla E, Younger DS, Chang HW, Tantravahi U, Miranda AF, Medori R, DiMauro S, Warburton D, Rowland LP: Partial dystrophin deficiency in monozygous twin carriers of the Duchenne gene discordant for clinical myopathy. Neurology 1990, 40(8):1267-1270.
• [14]Richards CS, Watkins SC, Hoffman EP, Schneider NR, Milsark IW, Katz KS, Cook JD, Kunkel LM, Cortada JM: Skewed X inactivation in a female MZ twin results in Duchenne muscular dystrophy. Am J Hum Genet 1990, 46(4):672-681.
• [15]Pegoraro E, Schimke RN, Arahata K, Hayashi Y, Stern H, Marks H, Glasberg MR, Carroll JE, Taber JW, Wessel HB, Bauserman SC, Marks WA, Toriello HV, Higgins JV, Appleton S, Schwartz L, Garcia CA, Hoffman EP: Detection of new paternal dystrophin gene mutations in isolated cases of dystrophinopathy in females. Am J Hum Genet 1994, 54(6):989-1003.
• [16]Azofeifa J, Voit T, Hübner C, Cremer M: X-chromosome methylation in manifesting and healthy carriers of dystrophinopathies: concordance of activation ratios among first degree female relatives and skewed inactivation as cause of the affected phenotypes. Hum Genet 1995, 96(2):167-176.
• [17]Bushby KM, Goodship JA, Nicholson LV, Johnson MA, Haggerty ID, Gardner-Medwin D: Variability in clinical, genetic and protein abnormalities in manifesting carriers of Duchenne and Becker muscular dystrophy. Neuromuscul Disord 1993, 3(1):57-64.
• [18]Sumita DR, Vainzof M, Campiotto S, Cerqueira AM, Cánovas M, Otto PA, Passos-Bueno MR, Zatz M: Absence of correlation between skewed X inactivation in blood and serum creatine-kinase levels in Duchenne/Becker female carriers. Am J Med Genet 1998, 80(4):356-361.
• [19]Soltanzadeh P, Friez MJ, Dunn D, von Niederhausern A, Gurvich OL, Swoboda KJ, Sampson JB, Pestronk A, Connolly AM, Florence JM, Finkel RS, Bönnemann CG, Medne L, Mendell JR, Mathews KD, Wong BL, Sussman MD, Zonana J, Kovak K, Gospe SM, Gappmaier E, Taylor LE, Howard MT, Weiss RB, Flanigan KM: Clinical and genetic characterization of manifesting carriers of DMD mutations. Neuromuscul Disord 2010, 20(8):499-504.
• [20]Seemann N, Selby K, McAdam L, Biggar D, Kolski H, Goobie S, Yoon G, Campbell C, Canadian Pediatric Neuromuscular Group: Symptomatic dystrophinopathies in female children. Neuromuscul Disord 2011, 21(3):172-7.
• [21]Matthews PM, Benjamin D, Van Bakel I, Squier MV, Nicholson LV, Sewry C, Barnes PR, Hopkin J, Brown R, Hilton-Jones D, Boyd Y, Karpati G, Brown GK, Craig IW: Muscle X-inactivation patterns and dystrophin expression in Duchenne muscular dystrophy carriers. Neuromuscul Disord 1995, 5(3):209-220.
• [22]Schwartz M, Duno M: Improved molecular diagnosis of dystrophin gene mutations using the multiplex ligation-dependent probe amplification method. Genet Test 2004, 8(4):361-367.
• [23]Gualandi F, Rimessi P, Trabanelli C, Spitali P, Neri M, Patarnello T, Angelini C, Yau SC, Abbs S, Muntoni F, Calzolari E, Ferlini A: Intronic breakpoint definition and transcription analysis in DMD/BMD patients with deletion/duplication at the 5' mutation hot spot of the dystrophin gene. Gene 2006, 370:26-33.
• [24]Bovolenta M, Neri M, Fini S, Fabris M, Trabanelli C, Venturoli A, Martoni E, Bassi E, Spitali P, Brioschi S, Falzarano MS, Rimessi P, Ciccone R, Ashton E, McCauley J, Yau S, Abbs S, Muntoni F, Merlini L, Gualandi F, Ferlini A: A novel custom high density-comparative genomic hybridization array detects common rearrangements as well as deep intronic mutations in dystrophinopathies. BMC Genomics 2008, 9:572-582. BioMed Central Full Text
• [25]Amos-Landgraf JM, Cottle A, Plenge RM, Friez M, Schwartz CE, Longshore J, Willard HF: X chromosome-inactivation patterns of 1,005 phenotypically unaffected females. Am J Hum Genet 2006, 79(3):493-499.
• [26]Brown R, Jarvis K, Hyland K: Protein measurement using bicinchoninic acid: elimination of interfering substances. Anal Biochem 1989, 180:136-139.
• [27]Hoogerwaard EM, Ginjaar IB, Bakker E, de Visser M: Dystrophin analysis in carriers of Duchenne and Becker muscular dystrophy. Neurology 2005, 65(12):1984-1986.
• [28]Pegoraro E, Schimke RN, Garcia C, Stern H, Cadaldini M, Angelini C, Barbosa E, Carroll J, Marks WA, Neville HE, Marks H, Appleton S, Toriello H, Wessel HB, Donnelly J, Bernes SM, Taber JW, Weiss L, Hoffman EP: Genetic and biochemical normalization in female carriers of Duchenne muscular dystrophy: evidence for failure of dystrophin production in dystrophin-competent myonuclei. Neurology 1995, 45(4):677-690.
• [29]Muntoni F, Mateddu A, Marrosu MG, Cau M, Congiu R, Melis MA, Cao A, Cianchetti C: Variable dystrophin expression in different muscles of a Duchenne muscular dystrophy carrier. Clin Genet 1992, 42(1):35-38.
• [30]Orstavik KH, Scheibel E, Ingerslev J, Schwartz M: Absence of correlation between X chromosome inactivation pattern and plasma concentration of factor VIII and factor IX in carriers of haemophilia A and B. Thromb Haemost 2000, 83(3):433-7.
• [31]Maier EM, Osterrieder S, Whybra C, Ries M, Gal A, Beck M, Roscher AA, Muntau AC: Disease manifestations and X inactivation in heterozygous females with Fabry disease. Acta Paediatr Suppl 2006, 95(451):30-8.
• [32]Kristiansen M, Knudsen GP, Tanner SM, McEntagart M, Jungbluth H, Muntoni F, Sewry C, Gallati S, Ørstavik KH, Wallgren-Pettersson C: X-inactivation patterns in carriers of X-linked myotubular myopathy. Neuromuscul Disord 2003, 13(6):468-71.
• [33]Pegoraro E, Hoffman EP, Piva L, Gavassini BF, Cagnin S, Ermani M, Bello L, Soraru G, Pacchioni B, Bonifati MD, Lanfranchi G, Angelini C, Kesari A, Lee I, Gordish-Dressman H, Devaney JM, McDonald CM, Cooperative International Neuromuscular Research Group: SPP1 genotype is a determinant of disease severity in Duchenne muscular dystrophy. Neurology 2011, 76(3):219-26.