Naturally occurring BRCA2 alternative mRNA splicing events in clinically relevant samples

被引:41
作者
Fackenthal, James D. [1 ]
Yoshimatsu, Toshio [1 ]
Zhang, Bifeng [1 ]
de Garibay, Gorka R. [1 ]
Colombo, Mara [2 ]
De Vecchi, Giovanna [2 ]
Ayoub, Samantha C. [1 ]
Lal, Kumar [1 ]
Olopade, Olufunmilayo I. [1 ]
Vega, Ana [3 ]
Santamarina, Marta [3 ]
Blanco, Ana [3 ]
Wappenschmidt, Barbara [4 ,5 ,6 ]
Becker, Alexandra [4 ,5 ,6 ]
Houdayer, Claude [7 ,8 ,9 ]
Walker, Logan C. [10 ]
Lopez-Perolio, Irene [11 ]
Thomassen, Mads [12 ]
Parsons, Michael [13 ]
Whiley, Phillip [13 ]
Blok, Marinus J. [14 ]
Brandao, Rita D. [15 ]
Tserpelis, Demis [14 ]
Baralle, Diana [16 ]
Montalban, Gemma [17 ,18 ]
Gutierrez-Enriquez, Sara [17 ,18 ]
Diez, Orland [17 ,18 ,19 ]
Lazaro, Conxi [20 ]
Spurdle, Amanda B. [13 ]
Radice, Paolo [2 ]
de la Hoya, Miguel [11 ]
机构
[1] Univ Chicago, Dept Med, Chicago, IL USA
[2] Fdn IRCCS Ist Nazl Tumori INT, Dept Prevent & Predict Med, Unit Mol Bases Genet Risk & Genet Testing, Milan, Italy
[3] IDIS, CIBERER, Grp Med Xenom USC, Fdn Publ Galega Med Xenom SERGAS, Santiago De Compostela, Spain
[4] Univ Cologne, Med Fac, Ctr Hereditary Breast & Ovarian Canc, CIO, Cologne, Germany
[5] Univ Cologne, CMMC, Cologne, Germany
[6] Univ Hosp Cologne, Cologne, Germany
[7] Inst Curie, Serv Genet, Paris, France
[8] Inst Curie, INSERM U830, Paris, France
[9] Univ Paris 05, Sorbonne Paris Cite, Paris, France
[10] Univ Otago, Dept Pathol, Christchurch, New Zealand
[11] Hosp Clin San Carlos, Inst Invest Sanitaria San Carlos IdISSC, Lab Oncol Mol, Madrid, Spain
[12] Odense Univ Hosp, Dept Clin Genet, Odense C, Denmark
[13] QIMR Berghofer Med Res Inst, Genet & Computat Biol Div, Brisbane, Qld, Australia
[14] Maastricht Univ, Dept Clin Genet, Med Ctr, Maastricht, Netherlands
[15] Maastricht Univ, Fac Humanities & Sci, Maastricht Sci Programme, Maastricht, Netherlands
[16] Univ Southampton, Human Dev & Hlth Acad Unit, Southampton Gen Hosp, Fac Med, Southampton, Hants, England
[17] VHIO, Oncogenet Grp, Barcelona, Spain
[18] Univ Autonoma Barcelona, Barcelona, Spain
[19] Univ Hosp Vall dHebron, Clin & Mol Genet Area, Barcelona, Spain
[20] IDIBELL Catalan Inst Oncol, Hereditary Canc Program, Mol Diagnost Unit, Barcelona, Spain
[21] Peter MacCallum Canc Ctr, East Melbourne, Vic, Australia
基金
澳大利亚国家健康与医学研究理事会;
关键词
SEQUENCE VARIANTS; UNCERTAIN SIGNIFICANCE; UNCLASSIFIED VARIANTS; FUNCTIONAL ASSAYS; DNA VARIANTS; MUTATIONS; CLASSIFICATION; RISK; GENE; TRANSCRIPTION;
D O I
10.1136/jmedgenet-2015-103570
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
Background BRCA1 and BRCA2 are the two principal tumour suppressor genes associated with inherited high risk of breast and ovarian cancer. Genetic testing of BRCA1/2 will often reveal one or more sequence variants of uncertain clinical significance, some of which may affect normal splicing patterns and thereby disrupt gene function. mRNA analyses are therefore among the tests used to interpret the clinical significance of some genetic variants. However, these could be confounded by the appearance of naturally occurring alternative transcripts unrelated to germline sequence variation or defects in gene function. To understand which novel splicing events are associated with splicing mutations and which are part of the normal BRCA2 splicing repertoire, a study was undertaken by members of the Evidence-based Network for the Interpretation of Germline Mutant Alleles (ENIGMA) consortium to characterise the spectrum of naturally occurring BRCA2 mRNA alternate-splicing events. Methods mRNA was prepared from several blood and breast tissue-derived cells and cell lines by contributing ENIGMA laboratories. cDNA representing BRCA2 alternate splice sites was amplified and visualised using capillary or agarose gel electrophoresis, followed by sequencing. Results We demonstrate the existence of 24 different BRCA2 mRNA alternate-splicing events in lymphoblastoid cell lines and both breast cancer and non-cancerous breast cell lines. Conclusions These naturally occurring alternate-splicing events contribute to the array of cDNA fragments that may be seen in assays for mutation-associated splicing defects. Caution must be observed in assigning alternate-splicing events to potential splicing mutations.
引用
收藏
页码:548 / 558
页数:11
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