title: Severe HDL deficiency due to novel defects in the ABCA1 transporter.
abstract: OBJECTIVE
  The objective was the identification and functional characterization of mutations in the ABCA1 gene in four patients with severe HDL deficiency.
  
  
  METHODS
  Patients were referred to the clinic because of almost complete HDL deficiency.
  
  
  METHODS
  The ABCA1 gene was sequenced directly. The analysis of the ABCA1 protein, ABCA1 mRNA and ABCA1-mediated cholesterol efflux was performed in cultured fibroblasts. Intracellular localization of ABCA1 mutants was investigated in transfected HEK293 cells.
  
  
  RESULTS
  Two patients were homozygous for mutations in the coding region of the ABCA1 gene, resulting in an amino acid substitution (p.A1046D) and a truncated protein (p.I74YFsX76). The third patient was homozygous for a splice site mutation in intron 35 (c.4773 + 1g>a), resulting in an in-frame deletion of 25 amino acids (del p.D1567_K1591) in ABCA1. These patients had clinical manifestations of accumulation of cholesterol in the reticulo-endothelial system. The fourth patient, with preclinical atherosclerosis, was a compound heterozygote for two missense mutations (p.R587W/p.W1699C). ABCA1-mediated cholesterol efflux was abolished in fibroblasts from patients with p.A1046D and del p.D1567_K1591 mutants and in fibroblasts homozygous for p.R587W. A reduced ABCA1 protein content was observed in these cells, suggesting an increased intracellular degradation. The mutant p.W1699C was largely retained in the endoplasmic reticulum, when expressed in HEK293 cells.
  
  
  CONCLUSIONS
  The homozygotes for mutations which abolish ABCA1 function showed overt signs of involvement of the reticulo-endothelial system. This was not the case in the compound heterozygote for missense mutations, suggesting that this patient retains some residual ABCA1 function that reduces cholesterol accumulation in the reticulo-endothelial system.
authors.affiliation: Department of Internal Medicine, University of Genoa, Genoa, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Pisciotta
authors.forename: L
authors.initials: L
authors.lastname: Bocchi
authors.forename: C
authors.initials: C
authors.lastname: Candini
authors.forename: R
authors.initials: R
authors.lastname: Sallo
authors.forename: I
authors.initials: I
authors.lastname: Zanotti
authors.forename: T
authors.initials: T
authors.lastname: Fasano
authors.forename: A
authors.initials: A
authors.lastname: Chakrapani
authors.forename: T
authors.initials: T
authors.lastname: Bates
authors.forename: R
authors.initials: R
authors.lastname: Bonardi
authors.forename: A
authors.initials: A
authors.lastname: Cantafora
authors.forename: S
authors.initials: S
authors.lastname: Ball
authors.forename: G
authors.initials: G
authors.lastname: Watts
authors.forename: F
authors.initials: F
authors.lastname: Bernini
authors.forename: S
authors.initials: S
authors.lastname: Calandra
authors.forename: S
authors.initials: S
authors.lastname: Bertolini
mesh_terms: D064286:ATP Binding Cassette Transporter 1; D018528:ATP-Binding Cassette Transporters / Q000235:genetics*; D000328:Adult; D000368:Aged; D019943:Amino Acid Substitution; D016632:Apolipoprotein A-I / Q000235:genetics; D002648:Child; D002675:Child, Preschool; D002784:Cholesterol / Q000378:metabolism*; D004252:DNA Mutational Analysis; D005091:Exons / Q000235:genetics; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D016368:Frameshift Mutation; D006801:Humans; D008075:Lipoproteins, HDL / Q000172:deficiency*; D008297:Male; D008875:Middle Aged; D020125:Mutation, Missense / Q000235:genetics*
chemical_list: C577836:ABCA1 protein, human; D064286:ATP Binding Cassette Transporter 1; D018528:ATP-Binding Cassette Transporters; D016632:Apolipoprotein A-I; D008075:Lipoproteins, HDL; D002784:Cholesterol