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Megalencephalic leukoencephalopathy with subcortical cysts type 1 (MLC1) due to a homozygous deep intronic splicing mutation (c.895-226T>G) abrogated in vitro using an antisense morpholino oligonucleotide

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Abstract

Megalencephalic leukoencephalopathy with subcortical cysts is an autosomal recessive disease characterized by early onset macrocephaly; developmental delay; motor disability in the form of progressive spasticity and ataxia; seizures; cognitive decline; and characteristic magnetic resonance imaging findings. Mutations in two genes, MLC1 (22q13.33; 75 % of patients) or HEPACAM (11q24; 20 % of patients), are associated with the disease. We describe an adult MLC patient with moderate clinical symptoms. MLC1 cDNA analysis from lymphoblasts showed a strong transcript reduction and identified a 246-bp pseudoexon containing a premature stop codon between exons 10 and 11, due to a homozygous c.895-226 T>G deep-intronic mutation. This category of mutations is often overlooked, being outside of canonically sequenced genomic regions. The mutation c.895-226 T>G has a leaky effect on splicing leaving part of the full-length transcript. Its role on splicing was confirmed using a minigene assay and an antisense morpholinated oligonucleotide targeted to the aberrant splice site in vitro, which partially abrogated the mutation effect.

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Acknowledgments

We thank Dr. D. Coviello for the pSPL3 vector (Invitrogen) and Prof. Nicola Migone for critical discussion. We are indebted to the patient and family members who collaborated in the study. This work was supported by Compagnia di San Paolo, Progetto finalizzato Neuroscienze entitled “Identification of a novel gene responsible for a form of adult-onset autosomal dominant leukodystrophy.”

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Authors and Affiliations

  1. Department of Genetics, Biology and Biochemistry, University of Torino, via Santena, 19–10126, Torino, Italy

    Cecilia Mancini, Laura Scalzitti, Simona Cavalieri, Alessandro Brussino & Alfredo Brusco

  2. Department of Neuroscience, AOU San Giovanni Battista, Torino, Italy

    Giovanna Vaula

  3. S.C.d.U. Medical Genetics, A.O.U. San Giovanni Battista, Torino, Italy

    Simona Cavalieri & Alfredo Brusco

  4. Laboratory of Molecular Medicine, Ospedale Pediatrico Bambino Gesù, Rome, Italy

    Enrico Bertini & Chiara Aiello

  5. Unità Operativa Complessa di Neurologia, Ospedale di Ivrea, Ivrea, Italy

    Cinzia Lucchini

  6. Departments of Pathology and Laboratory Medicine, University of California, Los Angeles, David Geffen School of Medicine, Los Angeles, CA, 90095-1732, USA

    Richard A. Gatti

Authors
  1. Cecilia Mancini
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  2. Giovanna Vaula
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  3. Laura Scalzitti
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  4. Simona Cavalieri
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  7. Cinzia Lucchini
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  8. Richard A. Gatti
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  9. Alessandro Brussino
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  10. Alfredo Brusco
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Correspondence to Alfredo Brusco.

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Alessandro Brussino and Alfredo Brusco contributed equally to the work.

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Mancini, C., Vaula, G., Scalzitti, L. et al. Megalencephalic leukoencephalopathy with subcortical cysts type 1 (MLC1) due to a homozygous deep intronic splicing mutation (c.895-226T>G) abrogated in vitro using an antisense morpholino oligonucleotide. Neurogenetics 13, 205–214 (2012). https://doi.org/10.1007/s10048-012-0331-z

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  • DOI: https://doi.org/10.1007/s10048-012-0331-z

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