18 THE NATIONAL MEDICAL JOURNAL OF INDIA VOL. 19, NO. 1, 2006 HFE, hepcidin and ferroportin gene mutations are not present in Indian patients with primary haemochromatosis PRIYANKA SHUKLA, SANDEEP JULKA, EESH BHATIA, SUDEEP SHAH, AABHA NAGRAL, RAKESH AGGARWAL ABSTRACT Background. Primaryhaemochromatosisischaracterizedby ironoverloadinthebodytissues.Itiscommoninpopulationsof northernEuropeandescent.Insuchpopulations,85%–90%of patientswiththisdiseasehaveaC282Ymutationinthe HFE gene.InIndia,thediseaseisuncommonandthegeneticdefects associatedwithitareunknown.Wethereforelookedformuta- tionsinthe HFE andothergenesinvolvedinironmetabolismin Indianpatientswithprimaryhaemochromatosis. Methods.Fivepatients(includingabrother–sisterpair)with primaryhaemochromatosisdiagnosedonclinical,biochemical and histological findings were studied. Genomic DNA was analysedbysequencingforthepresenceofmutationsinallthe 6exonsofthe HFEgeneandforpreviouslydescribedmutations ingenesencodinghepcidinantimicrobialpeptideandferroportin. Results . No patient had the C282Y mutation. One had homozygous H63D mutation. No other mutation was found in any HFE exon. Two previously reported splice site mutations in the HFE gene (IVS3+1 G/T and IVS5+1 G/A) were not detected. Four of the 5 patients had an HFE splice site mutation (IVS2+4 T/C; homozygous 2, heterozygous 2); however, this change was as frequent in 29 healthy subjects (homozygous 9, heterozygous 7), and was present in only 1 of the sibling pair patients, indicating that this repre- sented a polymorphism. No patient had any of the previously described mutations in the genes for hepcidin and ferroportin. Conclusion. Ourpatientswithprimaryhaemochromatosis lackedmutationsinthe HFE , hepcidinand ferroportingenes. Furthergeneticanalysismayhelpidentifynovelmutations responsibleforprimaryhaemochromatosisinthesepatients. Natl Med J India 2006;19:20–3 INTRODUCTION Primary haemochromatosis is a disorder of iron metabolism characterized by excessive accumulation of iron in the body tissues and organs leading to multiorgan dysfunction, including liver cirrhosis, hepatoma, diabetes and cardiac disorders. It is the most common genetic disorder among individuals of northern European descent, affecting nearly 1 in 300 individuals. 1–3 In 1996, a candidate gene for hereditary haemochromatosis was identified on the short arm of chromosome 6 and named the HFE gene. 4 In European countries, the majority of patients with haemochromatosis (85%–90%) have a G to A mutation on exon 4 of the HFE gene, which leads to substitution of cysteine at amino acid position 282 by tyrosine (C282Y). 4,5 Another HFE gene mutation from histidine to aspartic acid at amino acid position 63 (H63D) has also been implicated; 6 however, this mutation causes haemochromatosis only in association with C282Y heterozygos- ity on the other allele. 7,8 Also, two splice site mutations IVS5+1 G/ A and IVS3+1 G/T have been shown to be associated with heredi- tary haemochromatosis; 9,10 these mutations affect expression of the HFE protein through an alteration in splicing of the HFE gene mRNA. In a few patients with phenotypic expression of haemochro- matosis, mutations in other genes, named as hepcidin antimicro- bial peptide (HAMP), 11 transferrin receptor-2 (TFR2), 12 SLC11A3 Sanjay Gandhi Postgraduate Institute of Medical Sciences, Rae Bareli Road, Lucknow 226014, Uttar Pradesh, India PRIYANKA SHUKLA, RAKESH AGGARWAL Department of Gastroenterology SANDEEP JULKA, EESH BHATIA Department of Endocrinology P.D. Hinduja National Hospital, Veer Savarkar Marg, Mahim, Mumbai 400016, Maharashtra, India SUDEEP SHAH Jaslok Hospital and Research Centre, Dr G. Deshmukh Marg, Mumbai 400026, Maharashtra, India AABHA NAGRAL Correspondence to RAKESH AGGARWAL; aggarwal.ra @gmail.com © The National Medical Journal of India 2006