Expression of a mutant human fibrillin allele upon a normal human or murine genetic background recapitulates a Marfan cellular phenotype

Z A Eldadah, T Brenn, H Furthmayr, H C Dietz

Research output: Contribution to journalArticlepeer-review

Abstract

The Marfan syndrome (MFS) is a connective tissue disorder inherited as an autosomal dominant trait and caused by mutations in the gene encoding fibrillin, a 350-kD glycoprotein that multimerizes to form extracellular microfibrils. It has been unclear whether disease results from a relative deficiency of wild-type fibrillin; from a dominant-negative effect, in which mutant fibrillin monomers disrupt the function of the wild-type protein encoded by the normal allele; or from a dynamic and variable interplay between these two pathogenetic mechanisms. We have now addressed this issue in a cell culture system. A mutant fibrillin allele from a patient with severe MFS was expressed in normal human and murine fibroblasts by stable transfection. Immunohistochemical analysis of the resultant cell lines revealed markedly diminished fibrillin deposition and disorganized microfibrillar architecture. Pulse-chase studies demonstrated normal levels of fibrillin synthesis but substantially reduced deposition into the extracellular matrix. These data illustrate that expression of a mutant fibrillin allele, on a background of two normal alleles, is sufficient to disrupt normal microfibrillar assembly and reproduce the MFS cellular phenotype. This underscores the importance of the fibrillin amino-terminus in normal microfibrillar assembly and suggests that expression of the human extreme 5' fibrillin coding sequence may be sufficient, in isolation, to produce an animal model of MFS. Lastly, this substantiation of a dominant-negative effect offers mutant allele knockout as a potential strategy for gene therapy.

Original languageEnglish
Pages (from-to)874-80
Number of pages7
JournalJournal of Clinical Investigation
Volume95
Issue number2
DOIs
Publication statusPublished - Feb 1995

Keywords

  • Alleles
  • Animals
  • Base Sequence
  • Clone Cells
  • Exons
  • Extracellular Matrix Proteins
  • Fibroblasts
  • Fluorescent Antibody Technique
  • Frameshift Mutation
  • Gene Expression
  • Genetic Therapy
  • Humans
  • Marfan Syndrome
  • Mice
  • Microfilament Proteins
  • Molecular Sequence Data
  • Oligodeoxyribonucleotides
  • Phenotype
  • Polymerase Chain Reaction
  • Skin
  • Transcription, Genetic
  • Transfection

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