Using long-read sequencing to diagnose rare neuromuscular disorders: a pilot study

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dc.contributor.authorMartin, Alexandria Brooke
dc.contributor.examiningcommitteeWright, Galen (Max Rady College of Medicine)
dc.contributor.examiningcommitteeLee, Jae-Hyeok (Biological Sciences)
dc.contributor.supervisorHartley, Jessica
dc.contributor.supervisorBelmonte, Mark
dc.date.accessioned2025-05-13T16:52:59Z
dc.date.available2025-05-13T16:52:59Z
dc.date.issued2025-03
dc.date.submitted2025-05-13T16:52:59Zen_US
dc.degree.disciplineBiological Sciences
dc.degree.levelBachelor of Science (B.Sc.)
dc.description.abstractMany individuals with suspected genetic neuromuscular disorders (NMDs) remain undiagnosed despite extensive conventional genetic testing, which generally employs short-read sequencing (SRS) methods. Long-read sequencing (LRS) is a recently introduced technology which offers improved genome coverage and variant detection missed in standard approaches. Therefore, this pilot study aimed to investigate the application of long-read whole genome sequencing (LR-WGS) with the PacBio Revio platform for five participants (four families) with undiagnosed NMDs to identify causal genetic variants. Retrospective participant chart reviews were first conducted to assess phenotype, inheritance patterns, and other clinical factors. Following this, a novel bioinformatics pipeline was developed to enhance variant annotation and prioritization by integrating LRS population data with established genomic population frequency databases and in-silico predictions of deleteriousness. Finally, manual filtration was performed to further refine variant prioritization based on the clinical insights gained from chart reviews. Candidate variants of interest were found in three of four families analyzed (75%), including strong candidates in the FLNC and DMD genes, and a potential candidate in the TTN gene requiring functional validation. This pilot validates the pipeline for analysis of LRS data and provides evidence that LR-WGS could offer improvements over traditional SRS approaches in diagnosing NMDs, particularly in detecting complex variants. Improving accurate molecular diagnosis ultimately leads to better clinical outcomes for affected patients, including personalized treatment strategies and interventions. Future directions include the study of additional participants with NMDs to allow for a more comprehensive evaluation of diagnostic yield and applicability across diverse genetic backgrounds.
dc.identifier.urihttp://hdl.handle.net/1993/39077
dc.language.isoeng
dc.subjectneuromuscular disorders
dc.subjectNMD
dc.subjectlong-read sequencing
dc.subjectLRS
dc.titleUsing long-read sequencing to diagnose rare neuromuscular disorders: a pilot study
local.author.affiliationFaculty of Science::Department of Biological Sciences
oaire.awardTitleNew Investigator Operating Grant
project.funder.identifierhttp://dx.doi.org/10.13039/100008794
project.funder.nameResearch Manitoba
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