Association of elevated homocysteine levels and Methylenetetrahydrofolate reductase (MTHFR) 1298 A > C polymorphism with Vitiligo susceptibility in Gujarat Shahnawaz D. Jadeja a , Mohmmad Shoab Mansuri a , Mala Singh a , Hima Patel a , Yogesh S. Marfatia b , Rasheedunnisa Begum a, * a Department of Biochemistry, Faculty of Science, The Maharaja Sayajirao University of Baroda, Vadodara, 390002, Gujarat, India b Department of Skin and VD, Medical College Baroda, Vadodara, 390002, Gujarat, India A R T I C L E I N F O Article history: Received 5 April 2017 Received in revised form 13 December 2017 Accepted 12 January 2018 Keywords: Vitiligo Homocysteine Vitamin B 12 MTHFR Genetic polymorphisms A B S T R A C T Background: Several studies have reported hyperhomocysteinemia in vitiligo patients, suggesting the potential role of elevated homocysteine levels in precipitating vitiligo. Objectives: We aimed to estimate homocysteine and vitamin B 12 levels, and to investigate the role of MTHFR 677 C > T and 1298 A > C polymorphisms in vitiligo susceptibility in Gujarat population. Methods: Homocysteine and vitamin B 12 levels were estimated in plasma of 55 vitiligo patients and 60 controls by Electrochemiluminescence immunoassay (ECLIA). Polymerase chain reaction- restriction fragment length polymorphism (PCR-RFLP) and amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) techniques were used to genotype MTHFR 677 C > T and 1298 A > C polymorphisms in 520 vitiligo patients and 558 controls. Results: Our results showed significantly elevated homocysteine levels (p = 0.0003) as well as significant decrease in vitamin B 12 levels (p = 0.0102) in vitiligo patients, as compared to controls. No significant difference in genotype and allele frequencies of MTHFR 677 C > T polymorphism was observed among patients and controls, however, the frequency of ‘CC’ genotype of MTHFR 1298 A > Cpolymorphism was significantly increased in patients as compared to controls (p = 0.0151). Analysis based on the type of vitiligo revealed a significant increase in ‘C’ allele of MTHFR 1298 A > C polymorphism in patients with generalized (p = 0.003) and active (p = 0.007) vitiligo as compared to controls. Both the polymorphisms of MTHFR were in low linkage disequilibrium (LD) and susceptible ‘TC’ haplotype was more frequently observed (p = 0.008) in vitiligo patients. Interestingly, elevated homocysteine levels were also positively correlated with MTHFR 1298 A > C polymorphism in vitiligo patients. Structure based in silico prediction revealed structural perturbations in MTHFR protein due to Ala222Val and Glu429Ala amino acid substitution. Conclusions: The present findings suggest that MTHFR 1298 A > C polymorphism and, altered homocysteine and vitamin B 12 levels might play a vital role in the precipitation of vitiligo. © 2018 Japanese Society for Investigative Dermatology. Published by Elsevier B.V. All rights reserved. 1. Introduction Vitiligo is one of the most common cosmetic disfigurement disorders caused due to loss of functional melanocytes from the epidermis [1]. The disease can affect individuals of any race or sex and manifests before the age of 20 years in approximately half of the patients [2]. Worldwide prevalence of vitiligo is about 0.06–2.28% of the population [3]. The etiology of vitiligo is complex, however, certain genetic predisposition factors and a number of potential precipitating events such as oxidative stress, autoimmunity, neurological factors etc. were appear to be involved [4,5]. Various pro-oxidants generated during melanin synthesis and intrinsic antioxidant defense mechanisms that are compro- mised in pathologic conditions make epidermal melanocytes more vulnerable to oxidative stress [6]. In addition, the oxidative stress acts as the triggering event in progressive melanocyte destruction Abbreviations: MTHFR, methylene tetrahydrofolate reductase; GV, generalized vitiligo; LV, localized vitiligo; AV, active vitiligo; SV, stable vitiligo; SNP, single nucleotide polymorphism; ARMS-PCR, amplification refractory mutation system- polymerase chain reaction; PCR-RFLP, polymerase chain reaction-restriction fragment length polymorphism; OR, odds ratio; ECLIA, electrochemiluminescence immunoassay. * Corresponding author. E-mail address: rasheedunnisab@yahoo.co.in (R. Begum). https://doi.org/10.1016/j.jdermsci.2018.01.003 0923-1811/ © 2018 Japanese Society for Investigative Dermatology. Published by Elsevier B.V. All rights reserved. Journal of Dermatological Science xxx (2018) xxx–xxx G Model DESC 3314 No. of Pages 11 Please cite this article in press as: S.D. Jadeja, et al., Association of elevated homocysteine levels and Methylenetetrahydrofolate reductase (MTHFR) 1298 A > C polymorphism with Vitiligo susceptibility in Gujarat, J Dermatol Sci (2018), https://doi.org/10.1016/j.jdermsci.2018.01.003 Contents lists available at ScienceDirect Journal of Dermatological Science journa l home page : www.jdsjournal.com