Factor H Competitor Generated by Gene Conversion Events Associates with Atypical Hemolytic Uremic Syndrome

J Am Soc Nephrol. 2018 Jan;29(1):240-249. doi: 10.1681/ASN.2017050518. Epub 2017 Oct 9.

Abstract

Atypical hemolytic uremic syndrome (aHUS), a rare form of thrombotic microangiopathy caused by complement pathogenic variants, mainly affects the kidney microvasculature. A retrospective genetic analysis in our aHUS cohort (n=513) using multiple ligation probe amplification uncovered nine unrelated patients carrying a genetic abnormality in the complement factor H related 1 gene (CFHR1) that originates by recurrent gene conversion events between the CFH and CFHR1 genes. The novel CFHR1 mutants encode an FHR-1 protein with two amino acid substitutions, L290S and A296V, converting the FHR-1 C terminus into that of factor H (FH). Next-generation massive-parallel DNA sequencing (NGS) analysis did not detect these genetic abnormalities. In addition to the CFHR1 mutant, six patients carried the previously uncharacterized CFH-411T variant. In functional analyses, the mutant FHR-1 protein strongly competed the binding of FH to cell surfaces, impairing complement regulation, whereas the CFH-411T polymorphism lacked functional consequences. Carriers of the CFHR1 mutation presented with severe aHUS during adulthood; 57% of affected women in this cohort presented during the postpartum period. Analyses in patients and unaffected carriers showed that FH plasma levels determined by the nonmutated chromosome modulate disease penetrance. Crucially, in the activated endothelial (HMEC-1) cell assay, reduced FH plasma levels produced by the nonmutated chromosome correlated inversely with impairment of complement regulation, measured as C5b-9 deposition. Our data advance understanding of the genetic complexities underlying aHUS, illustrate the importance of performing functional analysis, and support the use of complementary assays to disclose genetic abnormalities not revealed by current NGS analysis.

Keywords: CFHR1; complement; gene conversion; hemolytic uremic syndrome.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Atypical Hemolytic Uremic Syndrome / genetics*
  • Binding, Competitive
  • Blood Proteins / genetics*
  • Cell Line
  • Complement C3b Inactivator Proteins / genetics*
  • Complement Factor H / genetics*
  • Complement Factor H / metabolism
  • Complement Membrane Attack Complex / metabolism
  • Female
  • Gene Conversion*
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Male
  • Middle Aged
  • Mutation
  • Pedigree
  • Penetrance
  • Retrospective Studies
  • Sequence Analysis, DNA / methods
  • Young Adult

Substances

  • Blood Proteins
  • CFHR1 protein, human
  • Complement C3b Inactivator Proteins
  • Complement Membrane Attack Complex
  • factor H-related protein 1
  • Complement Factor H