GALNS gene expression profiling in Morquio A patients' fibroblasts

L. Carraresi, R. Parini, C. Filoni, A. Caciotti, G. Sersale, S. Tomatsu, C. Orlando, E. Zammarchi, R. Guerrini, M. A. Donati, A. Morrone

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Background: Quantification studies of mutated mRNAs have not been carried out on Morquio A patients. Such studies are very important for the determination of stability of premature termination codons (PTC) bearing transcripts in order to assess the appropriateness of introducing the newly developed therapeutic strategies such as "stop codon read-through therapy". Methods: This paper focuses on the study of the GALNS gene and mRNAs in two severe forms of Morquio A patients' fibroblasts with development of a new and rapid real-time RT-PCR for detection and quantification of absolute mRNA copy number. Results: We identified two new mutations c.385A > T (p.K129X) and c.899 - 1G > C) in Pt1 and a known splicing defect c.120 + 1G > A in Pt2. Using RT-PCR and real-time RT-PCR in Pt2 we detected low levels of mRNAs, suggesting its instability; in Pt1, we detected three aberrant mRNAs introducing premature stop codons, suggesting that both the c.385A > T and c.899 - 1G > C mutations produce mRNAs capable of escaping the nonsense-mediated decay (NMD) pathway. Conclusions: The development of a real-time RT-PCR assay allows to absolutely quantify the GALNS mRNAs carrying mutations that lead to PTCs bearing transcripts, which escape the NMD process and are potentially suitable for the new therapeutic approach.

Original languageEnglish
Pages (from-to)72-76
Number of pages5
JournalClinica Chimica Acta
Volume397
Issue number1-2
DOIs
Publication statusPublished - Nov 2008

Fingerprint

Gene Expression Profiling
Fibroblasts
Gene expression
Messenger RNA
Bearings (structural)
Real-Time Polymerase Chain Reaction
Nonsense Codon
Mutation
Factor IX
Terminator Codon
Assays
Therapeutics
Genes
Polymerase Chain Reaction
Defects

Keywords

  • GALNS expression profiling
  • GenBank Accession number, D17629
  • HUGO-approved gene symbol, GALNS
  • Morquio A
  • Mucopolysaccharidosis IVA (MPS IVA, Morquio type A) OMIM disorder/gene accession number, 253000
  • N-acetylgalactosamine-6-sulfate sulfatase E.C. number 3.1.6.4
  • Real-time RT-PCR

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry

Cite this

Carraresi, L., Parini, R., Filoni, C., Caciotti, A., Sersale, G., Tomatsu, S., ... Morrone, A. (2008). GALNS gene expression profiling in Morquio A patients' fibroblasts. Clinica Chimica Acta, 397(1-2), 72-76. https://doi.org/10.1016/j.cca.2008.07.021

GALNS gene expression profiling in Morquio A patients' fibroblasts. / Carraresi, L.; Parini, R.; Filoni, C.; Caciotti, A.; Sersale, G.; Tomatsu, S.; Orlando, C.; Zammarchi, E.; Guerrini, R.; Donati, M. A.; Morrone, A.

In: Clinica Chimica Acta, Vol. 397, No. 1-2, 11.2008, p. 72-76.

Research output: Contribution to journalArticle

Carraresi, L, Parini, R, Filoni, C, Caciotti, A, Sersale, G, Tomatsu, S, Orlando, C, Zammarchi, E, Guerrini, R, Donati, MA & Morrone, A 2008, 'GALNS gene expression profiling in Morquio A patients' fibroblasts', Clinica Chimica Acta, vol. 397, no. 1-2, pp. 72-76. https://doi.org/10.1016/j.cca.2008.07.021
Carraresi L, Parini R, Filoni C, Caciotti A, Sersale G, Tomatsu S et al. GALNS gene expression profiling in Morquio A patients' fibroblasts. Clinica Chimica Acta. 2008 Nov;397(1-2):72-76. https://doi.org/10.1016/j.cca.2008.07.021
Carraresi, L. ; Parini, R. ; Filoni, C. ; Caciotti, A. ; Sersale, G. ; Tomatsu, S. ; Orlando, C. ; Zammarchi, E. ; Guerrini, R. ; Donati, M. A. ; Morrone, A. / GALNS gene expression profiling in Morquio A patients' fibroblasts. In: Clinica Chimica Acta. 2008 ; Vol. 397, No. 1-2. pp. 72-76.
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AU - Parini, R.

AU - Filoni, C.

AU - Caciotti, A.

AU - Sersale, G.

AU - Tomatsu, S.

AU - Orlando, C.

AU - Zammarchi, E.

AU - Guerrini, R.

AU - Donati, M. A.

AU - Morrone, A.

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N2 - Background: Quantification studies of mutated mRNAs have not been carried out on Morquio A patients. Such studies are very important for the determination of stability of premature termination codons (PTC) bearing transcripts in order to assess the appropriateness of introducing the newly developed therapeutic strategies such as "stop codon read-through therapy". Methods: This paper focuses on the study of the GALNS gene and mRNAs in two severe forms of Morquio A patients' fibroblasts with development of a new and rapid real-time RT-PCR for detection and quantification of absolute mRNA copy number. Results: We identified two new mutations c.385A > T (p.K129X) and c.899 - 1G > C) in Pt1 and a known splicing defect c.120 + 1G > A in Pt2. Using RT-PCR and real-time RT-PCR in Pt2 we detected low levels of mRNAs, suggesting its instability; in Pt1, we detected three aberrant mRNAs introducing premature stop codons, suggesting that both the c.385A > T and c.899 - 1G > C mutations produce mRNAs capable of escaping the nonsense-mediated decay (NMD) pathway. Conclusions: The development of a real-time RT-PCR assay allows to absolutely quantify the GALNS mRNAs carrying mutations that lead to PTCs bearing transcripts, which escape the NMD process and are potentially suitable for the new therapeutic approach.

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