Abstract
Molecular characterization is important for an accurate diagnosis in hereditary spastic paraplegia (HSP). Mutations in the gene SPAST (SPG4) are the most common cause of autosomal dominant forms. We performed targeted next generation sequencing (NGS) in a SPAST-negative HSP sample. Forty-four consecutive HSP patients were recruited from an adult neurogenetics clinic in Sydney, Australia. SPAST mutations were confirmed in 17 subjects, and therefore 27 SPAST-negative patients were entered into this study. Patients were screened according to mode of inheritance using a PCR-based library and NGS (Roche Junior 454 sequencing platform). The screening panel included ten autosomal dominant (AD) and nine autosomal recessive (AR) HSP-causing genes. A genetic cause for HSP was identified in 25.9 % (7/27) of patients, including 1/12 classified as AD and 6/15 as AR or sporadic inheritance. Several forms of HSP were identified, including one patient with SPG31, four with SPG7 (with one novel SPG7 mutation) and two with SPG5 (including two novel CYP7B1 frameshift mutations). Additional clinical features were noted, including optic atrophy and ataxia for patients with SPG5 and ataxia and a chronic progressive external ophthalmoplegia-like phenotype for SPG7. This protocol enabled the identification of a genetic cause in approximately 25 % of patients in whom one of the most common genetic forms of HSP (SPG4) was excluded. Targeted NGS may be a useful method to screen for mutations in multiple genes associated with HSP. More studies are warranted to determine the optimal approach to achieve a genetic diagnosis in this condition.
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Acknowledgments
We would like to thank the patients participating in this study and the referring clinicians, including Drs. David Schultz, Janankan Ravindran, Jeffrey Blackie, Michael Katakar and Judy Spies. We would also like to thank pathologists Dr. C Smith and Professor PC Blumbergs (Institute of Medical and Veterinary Science, Adelaide, Australia) and Professor G Nicholson (Concord Repatriation General Hospital, Sydney, Australia) for his assistance with the genetic analysis. Dr. Kishore R. Kumar, Dr. Nicholas F. Blair and Dr. Christina Liang receive National Health and Medical Research Council of Australia (NHMRC) postgraduate scholarships. Professor Christine Klein is supported by grants from the Hermann and Lilly Schilling Foundation, the Federal Ministry of Education and Research of Germany (BMBF, 01GI0201) and the European Union (MEFOPA). Professor Carolyn M. Sue has received support from the Australian Brain Foundation and the HSP Research Foundation, and is a recipient of the NHMRC practitioner fellowship (App1008433).
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On behalf of all authors, the corresponding author states that there is no conflict of interest.
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All human and animal studies have been approved by the appropriate ethics committee and have therefore been performed in accordance with the ethical standards laid down in the 1964 Declaration of Helsinki and its later amendments. All persons gave their informed consent prior to their inclusion in the study. We have not included any details that might disclose the identity of the subjects in the study.
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K. R. Kumar and N. F. Blair contributed equally to the study.
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Kumar, K.R., Blair, N.F., Vandebona, H. et al. Targeted next generation sequencing in SPAST-negative hereditary spastic paraplegia. J Neurol 260, 2516–2522 (2013). https://doi.org/10.1007/s00415-013-7008-x
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DOI: https://doi.org/10.1007/s00415-013-7008-x