Combination of whole genome sequencing, linkage, and functional studies implicates a missense mutation in titin as a cause of autosomal dominant cardiomyopathy with features of left ventricular noncompaction

Research output: Contribution to journalArticle

Authors

  • Robert Hastings
  • Carin P. de Villiers
  • Charlotte Hooper
  • Liz Ormondroyd
  • Alistair Pagnamenta
  • Stefano Lise
  • Silvia Salatino
  • Samantha J. L. Knight
  • Jenny C. Taylor
  • Kate L. Thomson
  • Linda Arnold
  • Spyros D. Chatziefthimiou
  • Petr V. Konarev
  • Matthias Wilmanns
  • Elisabeth Ehler
  • Andrea Ghisleni
  • Mathias Gautel
  • Edward Blair
  • Hugh Watkins

Colleges, School and Institutes

Abstract

BACKGROUND: High throughput next-generation sequencing techniques have made whole genome sequencing accessible in clinical practice; however, the abundance of variation in the human genomes makes the identification of a disease-causing mutation on a background of benign rare variants challenging.

METHODS AND RESULTS: Here we combine whole genome sequencing with linkage analysis in a 3-generation family affected by cardiomyopathy with features of autosomal dominant left ventricular noncompaction cardiomyopathy. A missense mutation in the giant protein titin is the only plausible disease-causing variant that segregates with disease among the 7 surviving affected individuals, with interrogation of the entire genome excluding other potential causes. This A178D missense mutation, affecting a conserved residue in the second immunoglobulin-like domain of titin, was introduced in a bacterially expressed recombinant protein fragment and biophysically characterized in comparison to its wild-type counterpart. Multiple experiments, including size exclusion chromatography, small-angle x ray scattering, and circular dichroism spectroscopy suggest partial unfolding and domain destabilization in the presence of the mutation. Moreover, binding experiments in mammalian cells show that the mutation markedly impairs binding to the titin ligand telethonin.

CONCLUSIONS: Here we present genetic and functional evidence implicating the novel A178D missense mutation in titin as the cause of a highly penetrant familial cardiomyopathy with features of left ventricular noncompaction. This expands the spectrum of titin's roles in cardiomyopathies. It furthermore highlights that rare titin missense variants, currently often ignored or left uninterpreted, should be considered to be relevant for cardiomyopathies and can be identified by the approach presented here.

Bibliographic note

© 2016 The Authors.

Details

Original languageEnglish
Pages (from-to)426-435
Number of pages10
JournalCirculation: Cardiovascular Genetics
Volume9
Issue number5
Early online date13 Sep 2016
Publication statusPublished - 1 Oct 2016

Keywords

  • Adult, Aged, Aged, 80 and over, Animals, COS Cells, Cercopithecus aethiops, Computational Biology, Connectin/chemistry, DNA Mutational Analysis/methods, Databases, Genetic, Echocardiography, Female, Genetic Linkage, Genetic Markers, Genetic Predisposition to Disease, Genome-Wide Association Study, Heredity, High-Throughput Nucleotide Sequencing, Humans, Isolated Noncompaction of the Ventricular Myocardium/diagnostic imaging, Male, Middle Aged, Models, Molecular, Mutation, Missense, Myocytes, Cardiac/metabolism, Pedigree, Phenotype, Polymorphism, Single Nucleotide, Protein Binding, Protein Conformation, Protein Stability, Rats, Risk Assessment, Risk Factors, Structure-Activity Relationship, Transfection, Young Adult