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Cited by 441Cystic Fibrosis Research AdvancesGenetic and Kidney Cyst DiseasesGenomics and Rare Diseases

A Novel Mutation in the Cystic Fibrosis Gene in Patients with Pulmonary Disease but Normal Sweat Chloride Concentrations

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Highsmith, W. Edward, Burch, Lauranell H., Zhou, Zhaoqing, Olsen, John C., Boat, Thomas E., Spock, Alexander, Gorvoy, Jack D., Quittell, Lynne +4 more·1994·New England Journal of Medicine
DOI 10.1056/nejm199410133311503
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Abstract

BACKGROUND: Many patients with chronic pulmonary disease similar to that seen in cystic fibrosis have normal (or nondiagnostic) sweat chloride values. It has been difficult to make the diagnosis of cystic fibrosis in these patients because no associated mutation in the cystic fibrosis transmembrane conductance regulator (CFTR) gene has been identified. METHODS: We evaluated 23 patients with pulmonary disease characteristic of cystic fibrosis but with sweat chloride concentrations in the normal range. Mutations in the CFTR gene were sought by direct sequencing of polymerase chain reaction-amplified nasal epithelial messenger RNA and by testing the functioning of affected epithelium. RESULTS: A cytidine phosphate guanosine dinucleotide C-to-T point mutation in intron 19 of the CFTR gene, termed 3849 + 10 kb C to T, was identified in 13 patients from eight unrelated families. This mutation was found in patients from three different ethnic groups with three different extended haplotypes. The mutation leads to the creation of a partially active splice site in intron 19 and to the insertion into most CFTR transcripts of a new 84-base-pair "exon," containing an in-frame stop codon, between exons 19 and 20. Normally spliced transcripts were also detected at a level approximately 8 percent of that found in normal subjects. This mutation is associated with abnormal nasal epithelial and sweat acinar epithelial function. CONCLUSIONS: We have identified a point mutation in intron 19 of CFTR and abnormal epithelial function in patients who have cystic fibrosis-like lung disease but normal sweat chloride values. The identification of this mutation indicates that this syndrome is a form of cystic fibrosis. Screening for the mutation should prove diagnostically useful in this population of patients.

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  • The 3849+10kbC>T CFTR mutation creates a novel donor splice site inside intron 22, leading to insertion of an 84-nucleotide cryptic exon.

    “The 3849+10kbC>T mutation creates a novel donor splice site inside intron 22 of the CFTR gene, leading to the insertion of the new cryptic exon of 84 nucleotides, which results in an in-frame stop codon and consequent production of a truncated non-functional CFTR protein32,33.”

    From Allele specific repair of splicing mutations in cystic fibrosis through AsCas12a genome editing (2019) · cites this paper as [32]

    The Genetic Analysis of Patients (p. 3) says the 3849+10 kb C-to-T substitution creates a sequence resembling a splice-donor site, and the Discussion (p. 4) describes an 84-base insert. The paper consistently places the mutation and insert in intron 19, not intron 22. (LLM, conf=0.90)

    Experimental — accuracy not yet measured

  • CFTR modulators (i.e. potentiator VX-770, Kalydeco, Vertex) are used in clinic also for CF patients carrying the splicing mutations of this study.

    “Even though CFTR modulators (i.e. potentiator VX-770, Kalydeco, Vertex) are used in clinic also for CF patients carrying the splicing mutations of this study”

    From Allele specific repair of splicing mutations in cystic fibrosis through AsCas12a genome editing (2019) · cites this paper as [32]

    The Discussion (p. 4) identifies the intron 19 CFTR mutation and shows that it causes alternative splicing, supporting the reference to patients carrying a splicing mutation studied in this paper. But the Discussion (p. 6) describes possible therapies as future alternatives; it does not report clinical use of VX-770/Kalydeco or other CFTR modulators in these patients. (LLM, conf=0.89)

    Experimental — accuracy not yet measured

  • CFTR modulators are associated with side effects and strongly depend on the residual correct splicing of mutant CFTR, which is extremely reduced and variable among patients.

    “these drugs are associated with side effects and strongly depend on the residual correct splicing of mutant CFTR, which is extremely reduced and variable among patients30–33.”

    From Allele specific repair of splicing mutations in cystic fibrosis through AsCas12a genome editing (2019) · cites this paper as [32]

    The Expression of Normally Spliced CFTR mRNA section (p. 4) reports that one patient with the intron 19 mutation had only 5–10% as much normally spliced CFTR mRNA as normal subjects. The paper does not establish that residual correct splicing varies among patients, or discuss CFTR modulator side effects or whether those drugs depend on residual splicing. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • The 3849+10kbC>T mutation results in an in-frame stop codon and truncated non-functional CFTR protein.

    “which results in an in-frame stop codon and consequent production of a truncated non-functional CFTR protein32,33.”

    From Allele specific repair of splicing mutations in cystic fibrosis through AsCas12a genome editing (2019) · cites this paper as [32]

    The Discussion (pp. 4–5) says the 3849+10kbC>T mutation causes an 84-bp insertion containing an in-frame stop codon, and that no full-length CFTR protein is expected from transcripts with the insert. It does not establish that a truncated, non-functional protein is produced; it also reports some normally spliced transcripts that could produce normal CFTR. (LLM, conf=0.87)

    Experimental — accuracy not yet measured

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Cites

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Cited by

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Authors
Highsmith, W. E., Burch, L. H., Zhou, Z., Olsen, J. C., Boat, T. E., Spock, A., Gorvoy, J. D., Quittell, L., Friedman, K. J., Silverman, L. M., Boucher, R. C., & Knowles, M. R.
Affiliations
  • University of North Carolina at Chapel Hill (US)
  • Cincinnati Children's Hospital Medical Center (US)
  • Duke University (US)
  • Long Island Jewish Medical Center (US)
  • NewYork–Presbyterian Hospital (US)
ORCID
Venue
New England Journal of Medicine
Publisher
Massachusetts Medical Society
Published
1994-10-13
Volume
331
Issue
15
Pages
974-980
ISSN
  • 0028-4793 (ISSN-L)
  • 1533-4406
DOI
10.1056/nejm199410133311503
OpenAlex
W2336764462
Cited by
441
References
1
Citations by year
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FWCI
27.50
Type
journal article
Language
en
Open access
Locations
  • New England Journal of Medicine (publishedVersion)
  • PubMed (publishedVersion)
  • Carolina Digital Repository (University of North Carolina at Chapel Hill) (submittedVersion)
  • UNC Libraries
Funders
  • National Heart, Lung, and Blood Institute (P01HL034322, HL-34322)
  • National Center for Research Resources (RR-000046, M01RR000046)
Topics
  • Health Sciences › Medicine › Pulmonary and Respiratory Medicine › Cystic Fibrosis Research Advances
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genetic and Kidney Cyst Diseases
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genomics and Rare Diseases
Keywords
cystic fibrosis, CFTR gene, splice site mutation, in-frame stop codon, extended haplotypes, mutation detection
MeSH terms
Adolescent, Adult, Base Sequence, Child, Child, Preschool, Chlorides, Chromosomes, Human, Pair 17, Cystic Fibrosis, Female, Humans, Introns, Lung Diseases, Obstructive, Male, Membrane Proteins, Molecular Sequence Data, Mutation, Nasal Mucosa, RNA, Messenger, Sweat, Polymerase Chain Reaction, Chloride Channels, Cystic Fibrosis Transmembrane Conductance Regulator

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