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Neurofibromatosis Type 1 pp 173–186Cite as

Deep Intronic NF1 Mutations and Possible Therapeutic Interventions

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Abstract

A significant proportion of germline mutations causing neurofibromatosis type 1 affect the correct splicing of the NF1 gene. Splicing is a complex mechanism by which introns from a pre-mRNA are removed. This process is finely regulated at the cellular level, and it has been shown that it can be modulated by using antisense oligonucleotides (AONs). Multiple studies have demonstrated the power of AONs in modulating abnormal splicing caused by constitutive DNA mutations, first in cultured cells, later in preclinical animal models, and currently in several promising clinical trials. A subset of splicing mutations, known as deep intronic mutations, create or strengthen acceptor or donor splice sites that, using a wild-type counterpart sequence, lead to the inclusion of a cryptic exon in the mRNA that will eventually produce an aberrant protein. In NF1, deep intronic mutations account for approximately 2 % of all NF1 germline mutations identified to date. Here we review different examples in which phosphorodiamidate morpholino oligomers (PMOs, a class of AONs) have been used to efficiently correct the abnormal splicing produced by NF1 deep intronic mutations. Reestablishment of normal splicing after PMO treatment has been analyzed at the mRNA level in different patient-derived primary cell cultures. Further, indirect evidence of neurofibromin function restoration has been assessed by quantifying the levels of active Ras before and after PMO treatment. In vitro results collected so far have demonstrated the considerable potential of this type of antisense therapy for deep intronic NF1-causing mutations, although further experimental studies, particularly in preclinical animal models, will be required in order to safely translate these results into clinical trials and eventually to the clinic. The success of clinical trials using AON technology for other mis-splicing mutations causing Duchenne muscular dystrophy (DMD) is encouraging and delineates a possible path from bench to bedside for these types of mutations causing neurofibromatosis type 1.

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Acknowledgements

We thank the patients who participated in this study for their willingness to contribute and to collaborate in all the proposed experiments. We thank Josep Biayna for critical advice and corrections to the manuscript as well as all the members of the ICO-IMPPC Neurofibromatosis working group. The authors wish to extend special thanks to the Asociación Española de Afectados de Neurofibromatosis for the grant awarded and for their constant support during the research. We would like also to thank the Asociación Española Contra el Cáncer, which recognized our group as one of the 2010 stable cancer research groups. Contract grant sponsor: Spanish Health Research Fund; Carlos III Health Institute; Catalan Health Institute and Autonomous Government of Catalonia. Contract grant numbers: ISCIIIRETIC: RD06/0020/1051, RD06/0020/1050, 2009SGR290, PI10/01422, CA08/00248.

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Authors and Affiliations

  1. Hereditary Cancer Program, Genetic Diagnostics Unit, Laboratori de Recerca Translacional, Institut Català d’Oncologia-ICO-IDIBELL, Hospital Duran i Reynals, Gran Via 199-203, L’Hospitalet de Llobregat, 08908, Spain

    Conxi Lázaro & Juana Fernández-Rodríguez

  2. Program on Hereditary Cancer, Institute of Predictive and Personalized Medicine of Cancer (IMPPC), Badalona, Barcelona, Spain

    Eduard Serra

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  1. Conxi Lázaro
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  2. Juana Fernández-Rodríguez
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  3. Eduard Serra
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Correspondence to Conxi Lázaro .

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Editors and Affiliations

  1. School of Medicine, Institute of Medical Genetics, Cardiff University, Heath Park Campus, Cardiff, 144XN, United Kingdom

    Meena Upadhyaya

  2. School of Medicine, Institute of Medical Genetics, Cardiff University, Heath Park, Cardiff, CF14 4XN, United Kingdom

    David N. Cooper

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Lázaro, C., Fernández-Rodríguez, J., Serra, E. (2012). Deep Intronic NF1 Mutations and Possible Therapeutic Interventions. In: Upadhyaya, M., Cooper, D. (eds) Neurofibromatosis Type 1. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-32864-0_13

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