HFE Genotype and Parameters of Iron Metabolism in German First-Time Blood Donors

Abstract

Background: Hemochromatosis is usually inherited in an autosomal recessive mode and associated with missense mutations in the hemochromatosis gene (HFE), an HLA class 1 related gene. However the degree of penetrance is presently matter of debate.

Methods: To elucidate the frequency of HFE mutations in a German population and the relationship between genotype and phenotype, we determined the HFE C282Y and H63D genotypes in 500 first-time blood donors using an allele-specific ligase chain reaction (LCR). Ferritin and transferrin saturation (TS) of all donors found to have at least one mutation were compared to gender- and age-matched controls.

Results: The C282Y allele frequency was 46 in 1000 chromosomes (4.6 %). The allele frequency of H63D was 108 in 1000 (10.8 %) chromosomes. We found three persons homozygous for H63D, nine compound heterozygotes and none homozygous for C282Y. TS was elevated in C282Y heterozygotes (p = 0.002) and C282Y/H63D compound heterozygotes (p = 0.04) compared to wild-type controls. Serum ferritin tended to be elevated in compound heterozygotes (p = 0.053). Mean corpuscular volume (MCV) and hemoglobin (MCH) were not different from controls.

Conclusion: The frequency of HFE mutations in the tested population was comparable to those of other northern European populations. The elevated TS in subjects carrying a single copy of the C282Y mutation suggests that C282Y heterozygosity is associated with an increased intestinal iron absorption and might therefore offer a selection advantage in conditions of iron depletion.

Zusammenfassung

Die hereditäre Hämochromatose ist eine autosomal rezessiv vererbbare Störung des Eisenstoffwechsels, die mit Mutationen im Hämochromatosegen (HFE), einem HLA-Klasse-1-ähnlichen Gen einhergeht. Das Ausmaß der Penetranz der Erkrankung wird gegenwärtig kontrovers diskutiert.

Methoden: 500 Erstblutspender wurden im Hinblick auf das Vorliegen der HFE-C282Y- und H63D-Mutation mithilfe einer allelspezifischen Ligasekettenreaktion untersucht. Ferritin und die Transferrinsättigung (TS) von Spendern, bei denen sich mindestens eine Mutation nachweisen ließ, wurden mit den Eisenstoffwechselparametern eines alters- und geschlechtsnormalisierten Kontrollkollektivs verglichen.

Ergebnisse: Die C282Y-Mutation wurde bei 46 von 1000 (4,6 %), die H63D-Mutation in 108 von 1000 (10,8 %) untersuchten Chromosomen festgestellt. Drei Spender waren H63D-homozygot, neun waren C282Y/H63D-compound-heterozygot. Keiner der Spender war C282Y-homozygot. Bei C282Y-heterozygoten und C282Y/H63D-compound-heterozygoten Individuen war die TS gegenüber den Kontrollkollektiven signifikant erhöht (p = 0,002 bzw. 0,04). Bei C282Y/H63D-compound-heterozygoten Spendern zeigte sich tendenziell ein erhöhter Ferritinspiegel (p = 0,053). Es zeigten sich keine Unterschiede im Hinblick auf das mittlere Erythrozytenvolumen (MCV) und den mittleren erythrozytären Hämoglobingehalt (MCH).

Diskussion: Die HFE-Allelfrequenz im untersuchten Kollektiv entspricht im Wesentlichen der anderer nordeuropäischer Populationen. Heterozygote Träger einer C282Y-Mutation haben eine erhöhte TS als Ausdruck einer vermehrten Eisenaufnahme. Dies ist möglicherweise Ausdruck eines Selektionsvorteils in Zeiten eines Eisenmangels.

Key words

Hemochromatosis - HFE - transferrin saturation - ferritin

Schlüsselwörter

Hämochromatose - HFE - Transferrinsättigung - Ferritin

Literature

• 1 Niederau C, Fischer R, Purschel A. et al . Long-term survival in patients with hereditary hemochromatosis.  Gastroenterology. 1996;  110 1107-1119
• 2 Feder J N, Gnirke A, Thomas W. et al . A novel MHC class I-like gene is mutated in patients with hereditary haemochromatosis.  Nat Genet. 1996;  13 399-408
• 3 Albig W, Drabent B, Burmester N. et al . The haemochromatosis candidate gene HFE (HLA-H) of man and mouse is located in syntenic regions within the histone gene cluster.  J Cell Biochem. 1998;  69 117-126
• 4 Bomford A. Genetics of haemochromatosis. Lancet; 2002 360: 1673-1681
• 5 Feder J N, Tsuchihashi Z, Irrinki A. et al . The hemochromatosis founder mutation in HLA-H disrupts beta2-microglobulin interaction and cell surface expression.  J Biol Chem. 1997;  272 14 025-14 028
• 6 Hohler T, Leininger S, Schneider P M. A new polymorphism in the human HFE gene.  Immunogenetics. 1999;  49 823-824
• 7 Mattman A, Huntsman D, Lockitch G. et al . Transferrin receptor 2 (TfR2) and HFE mutational analysis in non-C282Y iron overload: identification of a novel TfR2 mutation.  Blood. 2002;  100 1075-1077
• 8 Wallace D F, Pedersen P, Dixon J L. et al . Novel mutation in ferroportin1 is associated with autosomal dominant hemochromatosis.  Blood. 2002;  100 692-694
• 9 Roetto A, Papanikolaou G, Politou M. et al .Mutant antimicrobial peptide hepcidin is associated with severe juvenile hemochromatosis. Nat Genet 2003 33: 21-22
• 10 Ryan E, O’Keane C, Crowe J. Hemochromatosis in Ireland and HFE.  Blood Cells Mol Dis. 1998;  24 428-432
• 11 Cassanelli S, Pignatti E, Montosi G. et al . Frequency and biochemical expression of C282Y/H63D hemochromatosis (HFE) gene mutations in the healthy adult population in Italy.  J Hepatol. 2001;  34 523-528
• 12 Andrikovics H, Kalmar L, Bors A. et al . Genotype screening for hereditary hemochromatosis among voluntary blood donors in Hungary.  Blood Cells Mol Dis. 2001;  27 334-341
• 13 Gottschalk R, Seidl C, Loffler T. et al . HFE codon 63/282 (H63D/C282Y) dimorphism in German patients with genetic hemochromatosis.  Tissue Antigens. 1998;  51 270-275
• 14 Hellerbrand C, Bosserhoff A K, Seegers S. et al . Mutation analysis of the HFE gene in German hemochromatosis patients and controls using automated SSCP-based capillary electrophoresis and a new PCR-ELISA technique.  Scand J Gastroenterol. 2001;  36 1211-1216
• 15 Nielsen P, Carpinteiro S, Fischer R. et al . Prevalence of the C282Y and H63D mutations in the HFE gene in patients with hereditary haemochromatosis and in control subjects from Northern Germany.  Br J Haematol. 1998;  103 842-845
• 16 Merryweather C larke AT, Worwood M, Parkinson L. et al . The effect of HFE mutations on serum ferritin and transferrin saturation in the Jersey population.  Br J Haematol. 1998;  101 369-373
• 17 Jackson H A, Carter K, Darke C. et al . HFE mutations, iron deficiency and overload in 10,500 blood donors.  Br J Haematol. 2001;  114 474-484
• 18 Distante S, Berg J P, Lande K. et al . High prevalence of the hemochromatosis-associated Cys282Tyr HFE gene mutation in a healthy Norwegian population in the city of Oslo, and its phenotypic expression.  Scand J Gastroenterol. 1999;  34 529-534
• 19 Piperno A, Sampietro M, Pietrangelo A. et al . Heterogeneity of hemochromatosis in Italy.  Gastroenterology. 1998;  114 996-1002
• 20 De Gobbi M, Barilaro M R, Garozzo G. et al . TFR2 Y250X mutation in Italy.  Br J Haematol. 2001;  114 243-244
• 21 Erhardt A, Niederau C, Osman Y. et al . Nachweis des HFE-Polymorphismus bei deutschen Patienten mit hereditärer Hamochromatose. [Demonstration of HFE polymorphism in German patients with hereditary hemochromatosis].  Dtsch Med Wochenschr. 1999;  124 1448-1452
• 22 de Valk B, Witlox R S, van der Schouw Y T. et al . Biochemical expression of heterozygous hereditary hemochromatosis.  Eur J Intern Med. 2000;  11 317-321
• 23 Rossi E, Bulsara M K, Olynyk J K. et al . Effect of hemochromatosis genotype and lifestyle factors on iron and red cell indices in a community population.  Clin Chem. 2001;  47 202-208
• 24 Burke W, Thomson E, Khoury M J. et al .Hereditary hemochromatosis: gene discovery and its implications for population-based screening. JAMA 1998 280: 172-178
• 25 Beutler E. The HFE Cys282Tyr mutation as a necessary but not sufficient cause of clinical hereditary hemochromatosis.  Blood. 2003;  101 3347-3350
• 26 Beutler E, Felitti V J, Koziol J A. et al .Penetrance of 845G-> A (C282Y) HFE hereditary haemochromatosis mutation in the USA. Lancet; 2002 359: 211-218
• 27 Willis G, Wimperis J Z, Smith K C. et al .Haemochromatosis gene C282Y homozygotes in an elderly male population. Lancet; 1999 354: 221-222
• 28 Pietrangelo A. Hemochromatosis gene modifies course of hepatitis C viral infection.  Gastroenterology. 2003;  124 1509-1523
• 29 Erhardt A, Maschner-Olberg A, Mellenthin C. et al . HFE mutations and chronic hepatitis C: H63D and C282Y heterozygosity are independent risk factors for liver fibrosis and cirrhosis.  J Hepatol. 2003;  38 335-342
• 30 Bonkovsky H L, Jawaid Q, Tortorelli K. et al . Non-alcoholic steatohepatitis and iron: increased prevalence of mutations of the HFE gene in non-alcoholic steatohepatitis.  J Hepatol. 1999;  31 421-429
• 31 Chitturi S, Weltman M, Farrell G C. et al . HFE mutations, hepatic iron, and fibrosis: ethnic-specific association of NASH with C282Y but not with fibrotic severity.  Hepatology. 2002;  36 142-149
• 32 Egger N G, Goeger D E, Payne D A. et al . Porphyria cutanea tarda: multiplicity of risk factors including HFE mutations, hepatitis C, and inherited uroporphyrinogen decarboxylase deficiency.  Dig Dis Sci. 2002;  47 419-426
• 33 Dereure O, Aguilar M artinez P, Bessis D. et al . HFE mutations and transferrin receptor polymorphism analysis in porphyria cutanea tarda: a prospective study of 36 cases from southern France.  Br J Dermatol. 2001;  144 533-539
• 34 Tannapfel A, Stolzel U, Kostler E. et al . C282Y and H63D mutation of the hemochromatosis gene in German porphyria cutanea tarda patients.  Virchows Arch. 2001;  439 1-5
• 35 Gochee P A, Powell L W, Cullen D J. et al . A population-based study of the biochemical and clinical expression of the H63D hemochromatosis mutation.  Gastroenterology. 2002;  122 646-651
• 36 Bacon B R, Powell L W, Adams P C. et al . Molecular medicine and hemochromatosis: at the crossroads.  Gastroenterology. 1999;  116 193-207
• 37 Barton J C, Bertoli L F, Rothenberg B E. Screening for hemochromatosis in routine medical care: an evaluation of mean corpuscular volume and mean corpuscular hemoglobin.  Genet Test. 2000;  4 103-110
• 38 Datz C, Haas T, Rinner H. et al . Heterozygosity for the C282Y mutation in the hemochromatosis gene is associated with increased serum iron, transferrin saturation, and hemoglobin in young women: a protective role against iron deficiency?.  Clin Chem. 1998;  44 2429-2432
• 39 Beutler E, Felitti V, Gelbart T. et al . The effect of HFE genotypes on measurements of iron overload in patients attending a health appraisal clinic.  Ann Intern Med. 2000;  133 329-337
• 40 Beutler E, West C, Speir J A. et al . The hHFE gene of browsing and grazing rhinoceroses: a possible site of adaptation to a low-iron diet.  Blood Cells Mol Dis. 2001;  27 342-350