Abstract Details

Title Next Generation Sequencing in the Analysis of an Italian Cohort of Patients Affected by Nemaline Myopathy

Topic Muscle Disease/Neuromuscular Junction

Presentation(s) P07 - (-)

Poster/Presentation
Number 035

Objective

Background BACKGROUND: NM is a clinical and genetic heterogeneous disorder of skeletal muscle caused by mutations in five different genes encoding thin filament proteins of the striated muscle sarcomere. Mutations in NEB are responsible for about 50% of cases. 70 NEB mutations, mainly small deletions or point mutations, have been published. NEB is the largest and more complex gene among those involved in neuromuscular disorders, making its analysis by traditional sequencing method time consuming and expensive. Given the inherent gene complexity, we have chosen the NGS as an alternative technique.

Design/Methods DESIGN/METHODS: We have analysed 10 Italian patients with NM, previously screened for ACTA1, TPM2, TPM3, TNNT1 and CFL2. We have used the Target Region Capture Sequencing in order to investigate the NEB in depth, searching for mutations in all 183 exons, introns and promoter region. Alterations have been confirmed by Sanger.

Results RESULTS: We have found 9 novel NEB variants: missense mutations, affecting conserved amino acids; small indels causing frame shifts; nonsense mutations responsible for premature truncation of nebulin; splice-site mutations. In addition, we have found a deletion in the middle of intron 125 causing the partial inclusion of intron in the coding sequence.

Conclusions CONCLUSIONS: We have found mutations in 7 out of 10 patients, the highest detection rate observed in a selected cohort of NM patients. The application of the Target Region Capture Sequencing allowed a faster determination of recessive missense mutations, compound heterozygote genotypes, small deletions, stop codon and frame shifts. The target analysis has allowed to improve the quality and depth of data. We conclude that this method appears promising for a quicker detection of mutations in complex genes like NEB, with large coding size, exon number and without obvious mutational hotspots.

Authors/Disclosures
PRESENTER	No disclosure on file
	No disclosure on file
Francesca Magri (University of Milan; Department of Neurology)	No disclosure on file
Stefania Corti, MD, PhD (Foundation IRCCS Ca' Granda Ospedale Maggiore Policlinico)	Dr. Corti has received personal compensation in the range of $0-$499 for serving on a Scientific Advisory or Data Safety Monitoring board for Novartis. Dr. Corti has received personal compensation in the range of $0-$499 for serving on a Scientific Advisory or Data Safety Monitoring board for Biogen. Dr. Corti has received personal compensation in the range of $500-$4,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Sarepta.
Liana Apostolova, MD, FAAN (Indiana University School of Medicine)	Dr. Apostolova has received personal compensation in the range of $500-$4,999 for serving as a Consultant for NIH. Dr. Apostolova has received personal compensation in the range of $500-$4,999 for serving as a Consultant for Eli Lilly. Dr. Apostolova has received personal compensation in the range of $5,000-$9,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Roche. Dr. Apostolova has received personal compensation in the range of $500-$4,999 for serving on a Scientific Advisory or Data Safety Monitoring board for siemens. Dr. Apostolova has received personal compensation in the range of $5,000-$9,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Eisai. Dr. Apostolova has received personal compensation in the range of $5,000-$9,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Alnylam. Dr. Apostolova has received personal compensation in the range of $10,000-$49,999 for serving as an Editor, Associate Editor, or Editorial Advisory Board Member for Alzheimer Association. The institution of Dr. Apostolova has received research support from Roche Diagnostics. The institution of Dr. Apostolova has received research support from NIA. The institution of Dr. Apostolova has received research support from Alzheimer Association. The institution of Dr. Apostolova has received research support from AVID radiopharmaceuticals. The institution of Dr. Apostolova has received research support from Life Molecular Imaging. Dr. Apostolova has a non-compensated relationship as a advisor with FDA that is relevant to AAN interests or activities.
	No disclosure on file
	No disclosure on file
Enrico S. Bertini, MD (Bambino Gesu Hospital)	Dr. Bertini has received personal compensation for serving as an employee of Bambino Gesu' Children's Resesarch Hospital.
	No disclosure on file
	No disclosure on file
Maurizio Moggio, MD (University of Milan)	No disclosure on file
Maria D'Angelo, MD	No disclosure on file
	No disclosure on file
Marina Mora, PhD	No disclosure on file
Nereo Bresolin, DR	No disclosure on file
	No disclosure on file

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