SUMMARY:
This case highlights the utility of full-length isoform sequencing for characterizing the molecular mechanism of undiagnosed rare diseases and expands our understanding of the genetic basis for CMT2A.
TITLE:
Full-length isoform sequencing for resolving the molecular basis of Charcot-Marie-Tooth 2A
DESCRIPTION:
OBJECTIVES: Transcript sequencing of patient derived samples has been shown to improve the diagnostic yield for solving cases of likely Mendelian disorders, yet the added benefit of full-length long-read transcript sequencing is largely unexplored.
CONTENT:
bioRxiv. 2023 Feb 7:2023.02.07.526487. doi: 10.1101/2023.02.07.526487. Preprint.
ABSTRACT
OBJECTIVES: Transcript sequencing of patient derived samples has been shown to improve the diagnostic yield for solving cases of likely Mendelian disorders, yet the added benefit of full-length long-read transcript sequencing is largely unexplored.
METHODS: We applied short-read and full-length isoform cDNA sequencing and mitochondrial functional studies to a patient-derived fibroblast cell line from an individual with neuropathy that previously lacked a molecular diagnosis.
RESULTS: We identified an intronic homozygous MFN2 c.600-31T>G variant that disrupts a branch point critical for intron 6 spicing. Full-length long-read isoform cDNA sequencing after treatment with a nonsense-mediated mRNA decay (NMD) inhibitor revealed that this variant creates five distinct altered splicing transcripts. All five altered splicing transcripts have disrupted open reading frames and are subject to NMD. Furthermore, a patient-derived fibroblast line demonstrated abnormal lipid droplet formation, consistent with MFN2 dysfunction. Although correctly spliced full-length MFN2 transcripts are still produced, this branch point variant results in deficient MFN2 protein levels and autosomal recessive Charcot-Marie-Tooth disease, axonal, type 2A (CMT2A).
DISCUSSION: This case highlights the utility of full-length isoform sequencing for characterizing the molecular mechanism of undiagnosed rare diseases and expands our understanding of the genetic basis for CMT2A.
PMID:36798371 | PMC:PMC9934537 | DOI:10.1101/2023.02.07.526487
SOURCE:
bioRxiv : the preprint server for biology
TAGS:
CMT2A
CATEGORY:
Research
SUBCATEGORY:
n/a
DATE – PUBLISHED:
2023-02-08T05:15:20Z
DATE – DOI: 2023-02-08T05:15:20Z
DATE – PUBMED: 2023 Feb 7
DATE OUTPUT MATCHED: True
DATE – ADDED:
Fri, 17 Feb 2023 06:00:00 -0500
DATE – RETRIEVED:
02/17/23 07:07AM
2023-02-17T07:07:14-05:00
FEATURED IMAGE:
Media Uploaded (image/png)
IDENTIFIER:
pmid:36798371,pmc:PMC9934537,doi:10.1101/2023.02.07.526487
PUBMED ID:
pubmed:36798371
DOI:
10.1101/2023.02.07.526487
LINK – PUBMED:
https://pubmed.ncbi.nlm.nih.gov/36798371/
LINK – DOI:
https://doi.org/10.1101/2023.02.07.526487
LINK – PUBLISHER:
http://biorxiv.org/lookup/doi/10.1101/2023.02.07.526487
REFERENCES:
CMT Treatment Report, Urgent Research, 2023-02-17T07:07:14-05:00, https://www.cmttreatmentreport.com.
