Genetic study of variation in normal mouse iron homeostasis reveals ceruloplasmin as an HFE-hemochromatosis modifier gene.
OBJECTIVE
Genetic hemochromatosis is one of the most common genetic disorders, with progressive tissue iron overload leading to severe clinical complications. In Northern European populations, genetic hemochromatosis is usually caused by homozygosity for the C282Y mutation in the HFE protein. However, penetrance of this mutation is incomplete, suggesting that other genetic and environmental factors contribute to its differential biologic or clinical expression.


METHODS
To identify genes modifying iron homeostasis, we screened the 27 recombinant congenic strains of the C3H/DiSnA-C57BL/10ScSnA/Dem series for tissue and serum iron indices and genotyped 18 microsatellite markers in (C3H/DiSnA x HcB-2) F2 hybrid mice.


RESULTS
We identified 1 locus encompassing the Ceruloplasmin (Cp) gene with a strong linkage with liver iron, serum iron, and transferrin levels but not with spleen iron. Sequencing of Cp showed an R435X nonsense mutation in exon 7 in C3H/DiSnA mice. To evaluate whether Cp might act as a modifier gene of genetic hemochromatosis, we intercrossed C3H Hfe(-/-) and C3HDiSnA Cp(R435X/R435X) mice. As expected, we found that double-mutant mice deposited more iron in the liver than mice defective for either one or both genes. In contrast, Hfe(-/-) x Cp(R435/R435X) or Cp(R435X/R435X) x Hfe(+/-) showed 30% decrease in liver iron when compared with single mutant mice.


CONCLUSIONS
This study highlights the existence of complex interactions between Cp and HFE and represents the first example of a modifier gene with a protective effect, in which heterozygosity reduces the iron load in the context of HFE deficiency.
Gouya
Laurent
L
INSERM U773, Centre de Recherche Biomédicale Bichat Beaujon CRB3, Université Paris 7 Denis Diderot, site Bichat, Paris, France.
Muzeau
Francoise
F
Robreau
Anne-Marie
AM
Letteron
Philippe
P
Couchi
Eric
E
Lyoumi
Saïd
S
Deybach
Jean-Charles
JC
Puy
Herve
H
Fleming
Robert
R
Demant
Peter
P
Beaumont
Carole
C
Grandchamp
Bernard
B
D000818:Animals; D002570:Ceruloplasmin / Q000235:genetics*; D018389:Codon, Nonsense; D003433:Crosses, Genetic; D004195:Disease Models, Animal; D004912:Erythrocytes / Q000378:metabolism; D008040:Genetic Linkage; D005838:Genotype; D006432:Hemochromatosis / Q000097:blood / Q000235:genetics* / Q000378:metabolism* / Q000503:physiopathology; D000071020:Hemochromatosis Protein; D015395:Histocompatibility Antigens Class I / Q000235:genetics; D006706:Homeostasis / Q000235:genetics; D007501:Iron / Q000097:blood / Q000378:metabolism*; D008099:Liver / Q000378:metabolism; D008126:Lod Score; D008264:Macrophages / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000235:genetics; D051379:Mice; D020297:Mice, Congenic; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D018895:Microsatellite Repeats / Q000235:genetics; D008969:Molecular Sequence Data; D019683:Penetrance; D010587:Phagocytosis; D010641:Phenotype; D040641:Quantitative Trait Loci; D013154:Spleen / Q000378:metabolism; D014168:Transferrin / Q000378:metabolism
D018389:Codon, Nonsense; D000071020:Hemochromatosis Protein; C499535:Hfe protein, mouse; D015395:Histocompatibility Antigens Class I; D008565:Membrane Proteins; D014168:Transferrin; D007501:Iron; D002570:Ceruloplasmin