Introduction HFE gene is located on chromosome 6. Three missense mutations in HFE gene, C282Y, H63D, and S65C have been known to influence body iron status. 1 While being associated with haemochromatosis, their role (especially of HFE C282Y and HFE H63D) in the development of type 2 DM has been investigated in a number of Western populations. Moczulski et al. found high frequency of HFE 282Y allele in Polish subjects with type 2 DM compared to healthy controls and an association of HFE 63D allele with diabetic nephropathy. 2 Colli et al. reported association of HFE H63D polymorphism with high risk of type 2 DM in a Brazilian population. 3 Conversely, no association of HFE mutations with type 2 DM was found in populations belonging to United States and Turkey. 4,5 A meta-analysis of studies carried out between 1997- 2011 on the relationship of HFE polymorphism with the risk of type 2 DM showed no significant association of HFE C282Y with this disease. 6 However, carriers of HFE 63D allele had modestly increased risk of type 2 DM. 7 These reports were indicative of possible variation in the relationship of HFE polymorphism with type 2 DM from one population to another. Since no study had been carried out on South Asian populations to investigate the role of HFE polymorphism with type 2 DM, therefore, the objective of the present study was to find out the frequency distribution of 282Y, 63D, and 65C alleles in Pakistani subjects with type 2 DM and healthy controls and investigate if there is an association between genotypes of HFE C282Y, HFE H63D and HFE S65C polymorphisms with high body iron status and/or type 2 DM. The possibility of unique mutation(s) in HFE gene (other than the above mentioned 3 polymorphisms) in Pakistani subjects with type 2 DM having an association with high body iron status could not be discounted on the basis of the findings by Pointen et al. 8 Therefore, another objective of the study was to explore the presence of any J Pak Med Assoc 608 RESEARCH ARTICLE Lack of association of HFE gene polymorphism with high body iron status in Pakistani patients with type 2 diabetes mellitus Jibran Sualeh Muhammad, 1 Najmul Islam, 2 Naseema Mehboobali, 3 Khalida Iqbal, 4 Iqbal Azam, 5 Mohammad Perwaiz Iqbal 6 Abstract Objective: The Aim of this study was to investigate the relationship of 3 common polymorphisms in the HFE gene (C282Y, H63D and S65C) with high body iron status in a population of Pakistani subjects with type 2 diabetes mellitus (DM) and to explore if there is any novel mutation in HFE gene in a sample of Pakistani subjects with type 2 DM. Methods: In a case-control design, 200 healthy controls and 200 consecutive adult subjects with type 2 DM (both gender; age range of 30-70 years) were enrolled with informed consent. Their serum samples were analyzed for body iron status (ratio of concentration of soluble transferrin receptor to ferritin concentration). DNA from blood was screened for HFE gene polymorphisms via polymerase chain reaction, followed by restriction fragment length polymorphism or via Sanger sequencing to identify any novel mutation(s) in HFE gene. Results: We found that there was lack of any association between HFE polymorphism and body iron status in Pakistani subjects with type 2 DM and healthy controls. H63D was the most common polymorphism found in this population. Single base substitution of G nucleotide instead of C at the codon position 187 in the HFE gene exon 2 was discovered in one subject with DM. There was also a lack of association between D allele (variant allele of H63D) and type 2 DM. A significant relationship was found between CG genotype and abnormal albuminuria in subjects with type 2 DM (p = 0.036). Conclusion: In conclusion, HFE gene polymorphism is not associated either with high body iron status or type 2 DM in a hospital based Pakistani population and variant allele of H63D polymorphism appears to be associated with diabetic nephropathy. Keywords: High body iron; HFE gene, Polymorphism, Type 2 diabetes mellitus, Pakistani population, Albuminuria. (JPMA 71: 608; 2021) DOI: https://doi.org/10.47391/JPMA.563 1 Department of Basic Medical Sciences, College of Medicine, University of Sharjah, Sharjah, United Arab Emirates, 2 Department of Medicine, 3,4,6 Department of Biological and Biomedical Sciences, Medical College, 5 Department of Community Health Sciences, Aga Khan University, Karachi, Pakistan. Correspondence: Mohammad Perwaiz Iqbal. Email: Perwaiz.iqbal@aku.edu