Methylenetetrahydrofolate Reductase Activity Research Articles

Effect of L-Methylfolate Supplementation on Sleep for Patients with Reduced Methylenetetrahydrofolate Reductase Activity

Introduction Clinicians have limited options outside controlled substances to address sleep disturbance, which left untreated can negatively affect patient outcomes in cardiovascular health, mental health, immunologic function, and more. For some, genetic factors may influence sleep disturbances. L-methylfolate, the active form of folate, plays a critical role in regulation of monoamine neurotransmitters known to have significant impact on sleep regulation: dopamine, serotonin, norepinephrine. Single nucleotide polymorphisms of the enzyme methylene-tetrahydrofolate-reductase are common and can impact monoamine production. The goal of this study was to evaluate effects of L-methylfolate supplementation on sleep in a cohort with reduced methylene tetrahydrofolate reductase (MTHFR) activity. Methods A retrospective cohort of patients being treated with L-methylfolate in a concierge medical clinic setting was studied. Patients presenting with sleep complaints were evaluated using the Patient-Reported Outcomes Measurement Information System at baseline. Patients with known MTHFR polymorphisms at either C667T and/or A1298C were recommended 5 mg of L-methylfolate daily and were reevaluated at 2 wks, at 4 wks, and at 8 wks of supplementation. Statistical comparisons were made utilizing ANOVA and T-test comparisons. Results Ten were included in the final cohort: six male and four female, average age 43 ± 16 years. Beginning at wk 2, average sleep disturbance improved significantly by −6.94 points (p = 0.005) and by 8 wks, all patients had improvement with a −14.34 change in disturbance from baseline (p = 0.001). Conclusion Improvement in sleep disturbance was seen in both low and intermediate function phenotypes. L-methylfolate may be useful for improving sleep in patients with MTHFR polymorphism.

MTHFR C677T and A1298C polymorphism’s effect on risk of colorectal cancer in Lynch syndrome

Lynch syndrome (LS) is characterised by an increased risk of developing colorectal cancer (CRC) and other extracolonic epithelial cancers. It is caused by pathogenic germline variants in DNA mismatch repair (MMR) genes or the EPCAM gene, leading to a less functional DNA MMR system. Individuals diagnosed with LS (LS individuals) have a 10–80% lifetime risk of developing cancer. However, there is considerable variability in the age of cancer onset, which cannot be attributed to the specific MMR gene or variant alone. It is speculated that multiple genetic and environmental factors contribute to this variability, including two single nucleotide polymorphisms (SNPs) in the methylenetetrahydrofolate reductase (MTHFR) gene: C677T (rs1801133) and A1298C (rs1801131). By decreasing MTHFR activity, these SNPs theoretically reduce the silencing of DNA repair genes and increase the availability of nucleotides for DNA synthesis and repair, thereby protecting against early-onset cancer in LS. We investigated the effect of these SNPs on LS disease expression in 2,723 LS individuals from Australia, Poland, Germany, Norway and Spain. The association between age at cancer onset and SNP genotype (risk of cancer) was estimated using Cox regression adjusted for gender, country and affected MMR gene. For A1298C (rs1801131), both the AC and CC genotypes were significantly associated with a reduced risk of developing CRC compared to the AA genotype, but no association was seen for C677T (rs1801133). However, an aggregated effect of protective alleles was seen when combining the alleles from the two SNPs, especially for LS individuals carrying 1 and 2 alleles. For individuals with germline pathogenic variants in MLH1, the CC genotype of A1298C was estimated to reduce the risk of CRC significantly by 39% (HR = 0.61, 95% CI 0.42, 0.89, p = 0.011), while for individuals with pathogenic germline MSH2 variants, the AC genotype (compared to AA) was estimated to reduce the risk of CRC by 26% (HR = 0.66, 95% CI 0.53, 0.83, p = 0.01). In comparison, no association was observed for C677T (rs1801133). In conclusion, our study suggests that combining the MMR gene information with the MTHFR genotype, including the aggregated effect of protective alleles, could be useful in developing an algorithm that estimates the risk of CRC in LS individuals.

The importance of preconception Hcy testing: identification of a folate trap syndrome in a woman attending an assisted reproduction program.

Methylation is a ubiquitous and permanent key biochemical process playing a major role in gametogenesis and embryogenesis in relation to epigenetics and imprinting. Methylation relies on a unique cofactor S-Adenosyl Methionine: SAM. Release of the methyl group onto target molecules is followed by liberation of S-Adenosyl Homocysteine (SAH), and then homocysteine (Hcy), both potent inhibitors of the methylation process. Defective recycling of homocysteine, leading to Hyperhomocysteinemia, is mainly due to reduced activity of MTHFR (Methylene TetraHydroFolate Reductase). However, we described here, in a woman attending an ART program, arather rare syndrome: The Folate trap syndrome. Due to vitamin B12 deficiency (malabsorption), Hcy cannot be recycled to methionine by the methionine synthase. Transmethylation activity is weak and leads to Hhcy (Hyperhomocysteinhemia). Her Hhcy, over 16µM, was resistant to 5MTHF (5 Methyltetrahydrofolate) associated with a support of the one carbon cycle, a classical efficient treatment for elevated homocysteine. Treatment with Methylcobalamine (associated with adenosyl Cobalamine) allowed a Hcy drop down to 10 µM. Knowing the pleiotropic negative impact of Hcy on gametes, embryos and pregnancy in general, we strongly recommend a Hcy dosage in both members of couples seeking treatmentfor pregnancy.

No association between MTHFR gene C677T/A1298C polymorphisms, serum folate, vitamin B12, homocysteine levels, and prostate cancer in an Algerian population.

Methylenetetrahydrofolate reductase (MTHFR) is an important enzyme involved in folate and homocysteine metabolism, which are necessary for DNA methylation and nucleotide synthesis. Genetic polymorphisms that reduce MTHFR activity have been linked to several diseases, including prostate cancer. In this study, we aimed to investigate whether MTHFR polymorphisms, along with serum levels of folate, vitamin B12, and homocysteine, are associated with prostate cancer risk in the Algerian population. A total of 106 Algerian men with newly diagnosed prostate cancer and 125 healthy controls were included in this case-control study. The MTHFR C677T and A1298C polymorphisms were analyzed using PCR/RFLP and Real-Time PCR TaqMan® assays, respectively. Serum levels of folate, total homocysteine, and vitamin B12 were measured using an automatic biochemistry analyzer. We found no significant differences in the genotype frequency of A1298C and C677T between prostate cancer patients and controls. Moreover, serum levels of folate, total homocysteine, and vitamin B12 were not significantly associated with prostate cancer risk (p > 0.05). However, age and family history were identified as significant risk factors (OR = 1.178, p = 0.00 and OR = 10.03, p = 0.007, respectively). Our study suggests that MTHFR C677T and A1298C, as well as serum levels of folate, total homocysteine, and vitamin B12, are not associated with prostate cancer risk in the Algerian population. However, age and family history are significant risk factors. Further studies with a larger sample size are required to confirm these findings.

Folate-dependent hypermobility syndrome: A proposed mechanism and diagnosis

Hypermobility involves excessive flexibility and systemic manifestations of connective tissue fragility. We propose a folate-dependent hypermobility syndrome model based on clinical observations, and through a review of existing literature, we raise the possibility that hypermobility presentation may be dependent on folate status. In our model, decreased methylenetetrahydrofolate reductase (MTHFR) activity disrupts the regulation of the ECM-specific proteinase matrix metalloproteinase 2 (MMP-2), leading to high levels of MMP-2 and elevated MMP-2-mediated cleavage of the proteoglycan decorin. Cleavage of decorin leads ultimately to extracellular matrix (ECM) disorganization and increased fibrosis. This review aims to describe relationships between folate metabolism and key proteins in the ECM that can further explain the signs and symptoms associated with hypermobility, along with possible treatment with 5-methyltetrahydrofolate supplementation.

Th137Cs radionuclides, folate cycle and physical development of children living near the Chornobyl exclusion zone

The purpose of the study: to assess the impact of incorporated 137Cs radionuclides on the physical development of boys and girls living near the Chornobyl Exclusion Zone, taking into account the state of the folate cycle and the MTHFR:C677T genetic polymorphism. Research methods: іnstrumental, laboratory, mathematical and statistical. Results: Within the framework of international projects, an assessment of physical development was carried out according to the Rohrer index (IR), the levels and relationship of incorporated 137Cs, vitamins B6, B9, B12, homocysteine (Hcy), allelic variants of the MTHFR:C677T genetic polymorphism were determined in 260 children aged 13-17 years old from Ivankivskyi and Polisskyi districts of the Kyiv region, bordering the Chornobyl Exclusion Zone. It has been established that among the surveyed, the more proportion is the subgroup of adolescents with harmonious physical development (respectively, for boys - 70.40%, for girls - 65.93%). It was shown that boys and girls with disharmonious high physical development (respectively 15.20% and 25.19%) contained significantly less 137Cs radionuclides in their bodies, compared with children from the subgroups of harmonious and disharmonious low physical development. In the analyzed groups of boys and girls, an inverse correlation was recorded between the specific activity of 137Cs and the values of anthropometric indicators. The most pronounced inverse relationship between the specific activity of 137Cs and the values of mass, as well as body length and IR, was found in the subgroup of harmoniously developed girls. The subgroup of harmoniously developed boys differed from the subgroup of harmoniously developed girls by a higher specific activity of 137Cs in the body and a higher level of Hcy in the blood. The decrease in the level of Hcy in the blood of harmoniously developed girls, compared with harmoniously developed boys, may be associated with a higher level of vitamin B12 in the blood, as well as with a more intense functioning of the trans-sulfurization cycle. Comparative analysis showed that in the subgroup of girls with overweight, the proportion of cases of carriage of genotypes composed of neutral alleles of the C/C polymorphism MTHFR:677 is greater than in the subgroup of harmoniously developed girls. At the same time, a lower specific activity of 137Cs in their body was registered. It is concluded that incorporated 137Cs radionuclides cause a decrease in body weight in children living in the area affected by the accident at the Chоrnobyl nuclear power plant. At the same time, there is a metabolic disorder, in the form of an increase in the content of Hcy in the blood, more pronounced in boys. The process of incorporation of 137Cs radionuclides into the body of children may be associated with the T allele of the MTHFR:C677 polymorphism, which affects the activity of methylenetetrahydrofolate reductase.

Short-Term Combined Intake of Vitamin B2 and Vitamin E Decreases Plasma Homocysteine Concentrations in Female Track Athletes

Female athletes who endure physical strain on the same bone area are prone to stress fractures. Preventing these improves their quality of life. Blood vitamin B2 (V.B2) and E (V.E) levels influence blood homocysteine (Hcy) levels, which, when elevated, increase the risk of stress fractures. We aimed to determine the effects of V.B2 and V.E supplementation on plasma Hcy concentrations in these athletes carrying the C677T polymorphism in methylenetetrahydrofolate reductase (MTHFR). This study is a before–after study in 16 female athletes. It was divided into three intervention periods of V.B2 (28 mg/day), V.E (60 mg/day), and V.B2 + V.E intake, and each period was 14 days. Blood samples were collected before and after each period, and plasma Hcy concentration and each blood vitamin concentration were measured. Plasma Hcy concentration significantly decreased in all periods, but the maximum lowering effect was obtained when V.B2 and V.E were used in combination. The administration of V.B2 might have lowered the plasma Hcy level by maximizing the catalytic activity of MTHFR; V.E might have suppressed cell oxidation, increasing the efficiency of folate in lowering the plasma Hcy level. We propose the combined intake of V.B2 and V.E as effective nutrients to reduce plasma Hcy concentrations in female athletes with MTHFR polymorphisms.

Cerebral Venous Sinus Thrombosis with MTHFR C677T Heterozygous Mutation

Decrease in the activity of methylenetetrahydrofolate reductase (MTHFR) increases the level of homocysteine. MTHFR C677T gene polymorphism has been reported as a well-known cause of elevated homocysteine level associated with cerebral venous sinus thrombosis (CVST). A 34-year-old male was admitted with severe headache after generalized seizure. Brain magnetic resonance venography showed thrombus in superior sagittal sinus. In the investigation of risk factors, a slight increase in homocysteine caused by MTHFR C677T heterozygous mutation is presumed to be the cause of CVST.

High Folate, Perturbed One-Carbon Metabolism and Gestational Diabetes Mellitus.

Folate is a dietary micronutrient essential to one-carbon metabolism. The World Health Organisation recommends folic acid (FA) supplementation pre-conception and in early pregnancy to reduce the risk of fetal neural tube defects (NTDs). Subsequently, many countries (~92) have mandatory FA fortification policies, as well as recommendations for periconceptional FA supplementation. Mandatory fortification initiatives have been largely successful in reducing the incidence of NTDs. However, humans have limited capacity to incorporate FA into the one-carbon metabolic pathway, resulting in the increasingly ubiquitous presence of circulating unmetabolised folic acid (uFA). Excess FA intake has emerged as a risk factor in gestational diabetes mellitus (GDM). Several other one-carbon metabolism components (vitamin B12, homocysteine and choline-derived betaine) are also closely entwined with GDM risk, suggesting a role for one-carbon metabolism in GDM pathogenesis. There is growing evidence from in vitro and animal studies suggesting a role for excess FA in dysregulation of one-carbon metabolism. Specifically, high levels of FA reduce methylenetetrahydrofolate reductase (MTHFR) activity, dysregulate the balance of thymidylate synthase (TS) and methionine synthase (MTR) activity, and elevate homocysteine. High homocysteine is associated with increased oxidative stress and trophoblast apoptosis and reduced human chorionic gonadotrophin (hCG) secretion and pancreatic β-cell function. While the relationship between high FA, perturbed one-carbon metabolism and GDM pathogenesis is not yet fully understood, here we summarise the current state of knowledge. Given rising rates of GDM, now estimated to be 14% globally, and widespread FA food fortification, further research is urgently needed to elucidate the mechanisms which underpin GDM pathogenesis.

A Case-Control Study of the MTHFR C665T Gene Polymorphism on Macrocytic Anemia Among HIV-Infected Patients Receiving Zidovudine.

BackgroundMacrocytic anemia is the most common anemia in HIV-infected patients receiving zidovudine, and is closely related to folate and vitamin B12 deficiencies. Homocysteine >10 μmol/L and increased MMA (methylmalonic acid) levels >24.8 ng/mL indicate high/low folate and vitamin B12 deficiencies. Furthermore, MTHFR (Methylene-tetrahydrofolate-reductase) plays an essential role in the transmethylation of homocysteine to methionine and is related to DNA synthesis. The MTHFR C665T gene polymorphism decreases the activity of MTHFR, which culminates in homocysteinemia. Therefore, this case-control aims to assess the role of the MTHFR C665T gene polymorphism on the risk of macrocytic anemia among HIV-infected individuals receiving zidovudine.MethodsThis study was conducted using an unmatched case-control design and the participants were HIV-infected adults aged 20 to 59 years old, receiving zidovudine for four weeks and above. A sample of 232 patients was divided into case group with macrocytic anemia and the control having no anemia. Multivariate logistic regression analysis was then implemented to determine the risk factors.ResultsThe results showed that there was a significant difference in the number of female and male patients namely 51.3% and 48.7%, respectively, with p< 0.001. Moreover, the mean age of the cases and control group was 41.9 ± 9.4 and 36.2 ± 8.3. Regarding education, there were significant differences between subjects with low and high education 47.8% vs 52.2% with p<0.001. The majority of patients or 90.95% had taken AZT for more than 6 months. The logistic regression analysis test results showed that sex, age, education level, duration of AZT use, and homocysteine levels were predictors of macrocytic anemia with p<0.05, while MTHFR C665T gene polymorphism and MMA levels were not risk factors.ConclusionMTHFR C665T gene polymorphism does not contribute to the incidence of macrocytic anemia among HIV-infected individuals receiving zidovudine.

Decreased Methylenetetrahydrofolate Reductase Activity Leads to Increased Sensitivity to para-Aminosalicylic Acid in Mycobacterium tuberculosis.

ABSTRACTTuberculosis (TB), caused by Mycobacterium tuberculosis, is one of the most fatal diseases in the world. Methylenetetrahydrofolate reductase (MTHFR) catalyzes the production of 5-methyltetrahydrofolate (5-CH3-THF), which is required for the de novo biosynthesis of methionine in bacteria. Here, we identified Rv2172c as an MTHFR in M. tuberculosis through in vitro and in vivo analyses and determined that the protein is essential for the in vitro growth of the bacterium. Subsequently, we constructed rv2172c R159N and L214A mutants in M. tuberculosis and found that these mutants were more sensitive to the antifolates para-aminosalicylic acid (PAS) and sulfamethoxazole (SMX). Combining biochemical and genetic methods, we found that rv2172c R159N or L214A mutation impaired methionine production, leading to increased susceptibility of M. tuberculosis to PAS, which was largely restored by adding exogenous methionine. Moreover, overexpression of rv2172c in M. tuberculosis could increase methionine production and lead to PAS resistance. This research is the first to identify an MTHFR in M. tuberculosis and reveals that the activity of this enzyme is associated with susceptibility to antifolates. These findings have particular value for antitubercular drug design for the treatment of drug-resistant TB.

Methylenetetrahydrofolate Reductase Deficiency Reduces Brain Microglia in Zebrafish During Embryonic Development and Is Not Corrected by Folic Acid

ObjectivesNeuronal development and function is dependent on the interaction between the central nervous system and immune system. Microglia are resident macrophages of the brain critical for regulating neuronal activity during embryonic development. 5-methyltetrahydrofolate (5MTHF), the bioactive folate form, is essential for fetal brain development and immune function. Common variants in methylenetetrahydrofolate reductase (MTHFR), required for conversion of folic acid (FA) to 5-MTHF, limits its production. High dose FA supplementation is recommended but high FA may have the converse effect of reducing MTHFR activity. The objective of this study was to determine the effects of mthfr deficiency and its interaction with FA during embryonic development on microglia in a zebrafish model. MethodsThe mthfr gene in zebrafish was disrupted using two CRISPR mutagenesis methods. A set of 4 guide RNAs (gRNAs) + cas9 protein or cas9 alone (control) were injected to assay F0 zebrafish, or 2 gRNAs + cas9 mRNA were used to induce a germline mutation. To visualize macrophages at 4 days post fertilization (dpf) in live zebrafish, the transgenic mpeg1: mcherry line was used. In a subset of embryos, FA was added at 0, 50, 75, or 100- μM from 0–4dpf. At 4dpf, live neutral red staining for microglia was performed and the number in the optic tectum was quantified. 5MTHF, s-adenosylmethionine (SAM) and s-adenosylhomocysteine (SAH) were assayed in whole zebrafish at 5dpf. ResultsIn vivo imaging revealed a reduction in macrophage number (∼30%, P < 0.001) in the head region of mthfr disrupted zebrafish, but not in the periphery. mthfr zebrafish also had less microglia compared to controls (15%, P < 0.001). These changes were associated with lower 5MTHF (90%, P < 0.0001) and SAM: SAH (∼50%, P < 0.001) at 5dpf indicative of lower methylation potential. Exposure with FA did not correct the phenotype and at 100μM FA, control zebrafish also showed a decrease in microglia similar to mthfr zebrafish, confirming inhibitory effects of the high FA dose. Conclusionsmthfr deficiency reduces microglia in zebrafish but supplementation with FA does not prevent and may exacerbate the negative effects. The 5MTHF form of folate may be a better alternative to FA for brain health in patients with underlying genetic conditions. Funding SourcesSupported by CIHR-INMD.

Breast cancer prevention in districts affected by the Chernobyl nuclear power plant accident beginning from child ages

The aim of this paper was to assess the prevalence of the T risk allele of the MTHFR:677 genetic polymorphism in a group of girls from Ivankovsky and Polessky districts located near the Chernobyl exclusion zone. In addition, we assessed variants of combined carriership of the T allele with risk alleles of other genetic polymorphisms regulating the folate cycle. Research methods. Immunochemical, statistical. Results. Genetic predisposition to breast cancer risk was analyzed in a group of 251 adolescent girls. Carriership of the T allele of the MTHFR:С677Т polymorphism was found in 142 children (56.6%), while the homozygous T/T variant was found in 25 girls, or in 10.0% of cases. Compound heterozygosity for the 677CT/1298AC alleles of the MTHFR gene was recorded in 60 individuals, or in 23.9% of cases. Conclusions. The revealed genetic changes in the folate cycle lead to a significant decrease in the activity of methylenetetrahydrofolate reductase, and, accordingly, to an increase in the level of homocysteine in the blood, creating conditions for the occurrence of breast cancer. Given the high level of genetic predisposition, taking into account the constant impact on the body of radioactive elements and their decay products, the occurrence, as a consequence, of serious metabolic disorders, it is necessary to identify the breast cancer risk group of children.

Study of A1298C MTHFR Gene Polymorphism as a Risk Factor for Neural Tube Defects in the Eastern Algerian Population

Background: As C677T mutation, A1298C mutation in methylene tetrahydrofolate reductase (MTHFR) gene results in a decreased MTHFR activity but to a less extent, it is known as a risk factor of predisposition to human neural tube defects (NTDs), in some populations. Our objective was therefore to study, for the first time in Algerian population, if A1298C polymorphism confers risk for the occurrence of this abnormality. We have examined the distribution of the genotype and the allele frequencies of A1298C mutation, and also their influence on plasma homocysteine (Hcy) concentration. Patients and Methods: We studied this polymorphism in 38 mothers of NTD cases and 67 control individuals of an eastern Algerian population. The mutation was determined by polymerase chain reaction-restriction fragment length polymorphism analysis (PCR/RFLP). Plasma homocysteine concentration was analyzed using an automated chemiluminescence method. Results: No signi?cant association could be observed between allele and genotypes frequencies of A1298C MTHFR gene polymorphism and NTDs risk. However, we could observe that A1298C polymorphism affects homocysteine metabolism in mothers of NTD cases leading to homocysteine concentration values higher in AA genotype and lower in AC/CC genotypes (15.29 ± 11.8 μmol/l vs. 8.63 ± 3.83 μmol/l, p Conclusion: Data indicate that A1298C MTHFR gene polymorphism might be a risk factor by affecting homocysteine metabolism in mothers of Algerian children with NTDs.

Biochemical and clinical impact of reduced methylene tetrahydrofolate reductase enzyme activity

Methylene tetrahydrofolate reductase (MTHFR) enzyme is a crucial enzyme involved in the pathway of tetrahydrofolate inter-conversion and one-carbon metabolism. Though there are 24 reported polymorphisms associated to MTHFR gene, the two most investigated are C677T (Ala222 Val) and A1298C (Glu429 Ala) single nucleotide polymorphisms (SNPs). The metabolic impact in the form of hyperhomocysteinemia, redistribution of different forms of folates, altered methylation process and accumulation of unmetabolised folic acid have been reported in individuals with reduced or deficient MTHFR activity. These biochemical alterations have been associated with coronary artery diseases, various thromboembolic phenomenon, infertility and reproductive failures, cancers and inflammatory diseases. Folic acid supplementation may further worsen the case in these individuals by increasing the unmetabolised forms. Supplementation of 5-MTHF could be a better alternative for therapeutic and preventive purposes. Keywords: MTHFR, Structure, C677T, A1298C, Biochemical impact.

Embryonic MTHFR contributes to blastocyst development.

Reduction in methylenetetrahydrofolate reductase (MTHFR) activity due to genetic variations in the MTHFR gene has been controversially implicated in subfertility in human in vitro fertilization. However, there is no direct gene-knockdown study of embryonic MTHFR to assess its involvement in mammalian preimplantation development. The purpose of this study is to investigate expression profiles and functional roles of MTHFR in bovine preimplantation development. Reverse transcription-quantitative PCR (RT-qPCR) and analysis of publicly available RNA-seq data were performed to reveal expression levels of MTHFR during bovine preimplantation development. We knocked down MTHFR by siRNA-mediated RNA interference from the 8- to 16-cell stage and assessed the effects on preimplantation development. The RT-qPCR analysis showed relatively high MTHFR expression at the GV oocyte stage, which was decreased toward the 8- to 16-cell stage and then slightly restored at the blastocyst stage. Public data-based analysis also showed the similar pattern of expression with substantial embryonic expression at the blastocyst stage. MTHFR knockdown reduced the blastocyst rate (P < 0.01) and the numbers of total (P < 0.0001), trophectoderm (P < 0.0001), and inner cell mass (P < 0.001) cells. The results indicate that embryonic MTHFR is indispensable for normal blastocyst development. The findings provide insight into the debatable roles of MTHFR in fertility and may be applicable for the improvement of care for early embryos via modulation of surrounding folate-related nutritional conditions in vitro and/or in utero, depending on the parental and embryonic MTHFR genotype.

Methylene tetrahydrofolate reductase C677T polymorphism in Korean livedoid vasculopathy patients

To the Editor: Livedoid vasculopathy (LV) is a rare, chronic, ulcerative disease that significantly impairs the quality of life of patients.1Micieli R. Alavi A. Livedoid vasculopathy: an updated review.Curr Dermatol Rep. 2018; 7: 125-135Crossref Scopus (4) Google Scholar Previous case series have described the association between LV and various hypercoagulable conditions, including methylenetetrahydrofolate reductase (MTHFR) polymorphism and hyperhomocysteinemia.1Micieli R. Alavi A. Livedoid vasculopathy: an updated review.Curr Dermatol Rep. 2018; 7: 125-135Crossref Scopus (4) Google Scholar, 2Di Giacomo T.B. Hussein T.P. Souza D.G. Criado P.R. Frequency of thrombophilia determinant factors in patients with livedoid vasculopathy and treatment with anticoagulant drugs—a prospective study.J Eur Acad Dermatol Venereol. 2010; 24: 1340-1346Crossref PubMed Scopus (49) Google Scholar, 3Hairston B.R. Davis M.D. Pittelkow M.R. Ahmed I. Livedoid vasculopathy: further evidence for procoagulant pathogenesis.Arch Dermatol. 2006; 142: 1413-1418Crossref PubMed Scopus (148) Google Scholar This single-center, retrospective, case-control study included 43 patients with histologically confirmed LV between 2005 and 2019. Among them, hypercoagulable conditions were assessed in 28 patients who underwent 1 or more coagulation laboratory tests. A non-LV control group included 69 patients with psoriasis or eczema to compare MTHFR polymorphism and homocysteine level with patients with LV. Of 28 patients with LV, 10 (35.7%) showed 1 or more coagulation laboratory abnormalities (Supplemental Table I, available via Mendeley at https://doi.org/10.17632/ddghn6cdp6.2), which is a little lower than the 41.4% to 52% reported in Caucasian studies.2Di Giacomo T.B. Hussein T.P. Souza D.G. Criado P.R. Frequency of thrombophilia determinant factors in patients with livedoid vasculopathy and treatment with anticoagulant drugs—a prospective study.J Eur Acad Dermatol Venereol. 2010; 24: 1340-1346Crossref PubMed Scopus (49) Google Scholar,3Hairston B.R. Davis M.D. Pittelkow M.R. Ahmed I. Livedoid vasculopathy: further evidence for procoagulant pathogenesis.Arch Dermatol. 2006; 142: 1413-1418Crossref PubMed Scopus (148) Google Scholar In Caucasians, factor V Leiden mutation, prothrombin G20210A mutation, and antiphospholipid antibody presence were most common.1Micieli R. Alavi A. Livedoid vasculopathy: an updated review.Curr Dermatol Rep. 2018; 7: 125-135Crossref Scopus (4) Google Scholar, 2Di Giacomo T.B. Hussein T.P. Souza D.G. Criado P.R. Frequency of thrombophilia determinant factors in patients with livedoid vasculopathy and treatment with anticoagulant drugs—a prospective study.J Eur Acad Dermatol Venereol. 2010; 24: 1340-1346Crossref PubMed Scopus (49) Google Scholar, 3Hairston B.R. Davis M.D. Pittelkow M.R. Ahmed I. Livedoid vasculopathy: further evidence for procoagulant pathogenesis.Arch Dermatol. 2006; 142: 1413-1418Crossref PubMed Scopus (148) Google Scholar In contrast, none of these were detected in our patients. Instead, in our patients, MTHFR C677T TT homozygous polymorphism (29.2%) was the most frequent, followed by increased concentration of lipoprotein(a) (27.3%) and homocysteine (12.5%). C677T polymorphism, the most common polymorphism of MTHFR, which is an essential enzyme in homocysteine metabolism, would lead to the elevation of the plasma homocysteine level, and elevated homocysteine level is a well-known risk factor for atherosclerosis. LV has been reported to be associated with MTHFR C677T polymorphism or hyperhomocysteinemia, or both.1Micieli R. Alavi A. Livedoid vasculopathy: an updated review.Curr Dermatol Rep. 2018; 7: 125-135Crossref Scopus (4) Google Scholar, 2Di Giacomo T.B. Hussein T.P. Souza D.G. Criado P.R. Frequency of thrombophilia determinant factors in patients with livedoid vasculopathy and treatment with anticoagulant drugs—a prospective study.J Eur Acad Dermatol Venereol. 2010; 24: 1340-1346Crossref PubMed Scopus (49) Google Scholar, 3Hairston B.R. Davis M.D. Pittelkow M.R. Ahmed I. Livedoid vasculopathy: further evidence for procoagulant pathogenesis.Arch Dermatol. 2006; 142: 1413-1418Crossref PubMed Scopus (148) Google Scholar The univariable and multivariable logistic regression both showed that female sex and MTHFR C677T TT genotype were significantly associated with LV (Table I). However, 87.5% of patients with LV showed a homocysteine level within normal reference ranges. This implicates that MTHFR C677T polymorphism could increase the risk of LV regardless of the homocysteine level. In line with our findings, several case reports demonstrated MTHFR C677T TT homozygous polymorphism with normal homocysteine level in LV.1Micieli R. Alavi A. Livedoid vasculopathy: an updated review.Curr Dermatol Rep. 2018; 7: 125-135Crossref Scopus (4) Google Scholar In parallel, a study demonstrated that 47% of patients with vascular events had MTHFR polymorphism with a normal homocysteine level, and no other thrombophilia risk factors were identified.4Gadiyaram V.K. Khan M.A. Hogan T. et al.Significance of MTHFR gene mutation with normal homocysteine level in vascular events.J Clin Oncol. 2009; 27 (e20520-e)Crossref Google ScholarTable IAssociations between livedoid vasculopathy and different factorsVariable∗Data are presented as number (%) or as mean ± SD.Livedoid vasculopathy (n = 28)No livedoid vasculopathy (n = 69)Odds ratio (95% CI)CrudeAdjustedAge, y45.6 ± 15.143.8 ± 17.01.01 (0.98-1.03)…Female sex20 (71.4)25 (36.2)4.40 (1.69-11.44)†P < .01.5.18 (1.77-15.11)†P < .01.MTHFR C677T genotype‡Data were available except for 4 patients with LV. CC homozygote4 (16.6)25 (36.2)ReferenceReference CT heterozygote13 (54.2)36 (52.2)2.26 (0.66-7.73)3.33 (0.90-12.40) TT homozygote7 (29.2)8 (11.6)5.47 (1.27-23.64)§P < .05.6.87 (1.44-32.90)§P < .05.Homocysteine, μmol/LǁData were available except for 4 patients with LV and 2 controls.9.3 ± 4.18.9 ± 3.01.04 (0.90-1.19)…CI, Confidence interval; MTHFR, methylenetetrahydrofolate reductase.∗ Data are presented as number (%) or as mean ± SD.† P < .01.‡ Data were available except for 4 patients with LV.§ P < .05.ǁ Data were available except for 4 patients with LV and 2 controls. Open table in a new tab CI, Confidence interval; MTHFR, methylenetetrahydrofolate reductase. In fact, MTHFR C677T polymorphism is not the only factor that influences homocysteine level. Because the homocysteine metabolism by MTHFR requires folate and vitamin B12 as cofactors, supplementation of them can lower the homocysteine level and compensate the decreased activity of MTHFR.5Errichetti E. Stinco G. Recalcitrant livedoid vasculopathy associated with hyperhomocysteinaemia responding to folic acid and vitamins B6/B12 supplementation.Acta Derm Venereol. 2016; 96: 987-988Crossref PubMed Scopus (6) Google Scholar When patient characteristics of LV were compared against different MTHFR C677T genotypes, MTHFR polymorphism was not related with any patient characteristics (Table II). Theoretically, the MTHFR C677T polymorphism might be related only with the incidence of LV, not with further progression of the disease.Table IIComparison of patient characteristics of livedoid vasculopathy according to the methylenetetrahydrofolate reductase (MTHFR) C677T genotypesVariable∗The results are presented as mean ± SD or as median (range).MTHFR genotypesP valueCC homozygote (n = 4)CT heterozygote (n = 13)TT homozygote (n = 7)Onset age, y27.0 ± 9.640.8 ± 15.733.7 ± 14.8.244Disease duration, y9.8 ± 8.79.2 ± 8.07.0 ± 2.6.913Disease severity score (range 0-9)4.5 (3-6)5.3 (3-9)5 (3-8).499Homocysteine, μmol/L7.0 ± 2.88.6 ± 2.811.9 ± 5.5.105MTHFR, Methylenetetrahydrofolate reductase.∗ The results are presented as mean ± SD or as median (range). Open table in a new tab MTHFR, Methylenetetrahydrofolate reductase. This study has some limitations, including its retrospective nature and the small number of patients due to the rarity of LV. However, the present study is the largest study, to our knowledge, to demonstrate the associated hypercoagulable conditions in Korean patients with LV, as well as the first study worldwide to find that the MTHFR C677T polymorphism increases the risk of LV by comparison with non-LV control. In summary, more than one-third of Korean patients with LV had hypercoagulable abnormalities, of which MTHFR C677T polymorphism was the most frequent. Female sex and MTHFR C677T TT genotype were independent risk factors of LV. In contrast, the homocysteine level was normal in most patients.

Endothelial Dysfunction May Link Interatrial Septal Abnormalities and MTHFR-Inherited Defects to Cryptogenic Stroke Predisposition.

We explored the significance of the L-Arginine/asymmetric dimethylarginine (L-Arg/ADMA) ratio as a biomarker of endothelial dysfunction in stroke patients. To this aim, we evaluated the correlation, in terms of severity, between the degree of endothelial dysfunction (by L-Arg/ADMA ratio), the methylene tetrahydrofolate reductase (MTHFR) genotype, and the interatrial septum (IAS) phenotype in subject with a history of stroke. Methods and Results: L-Arg, ADMA, and MTHFR genotypes were evaluated; the IAS phenotype was assessed by transesophageal echocardiography. Patients were grouped according to the severity of IAS defects and the residual enzymatic activity of MTHFR-mutated variants, and values of L-Arg/ADMA ratio were measured in each subgroup. Of 57 patients, 10 had a septum integrum (SI), 38 a patent foramen ovale (PFO), and 9 an ostium secundum (OS). The L-Arg/ADMA ratio differed across septum phenotypes (p ≤ 0.01), and was higher in SI than in PFO or OS patients (p ≤ 0.05, p ≤ 0.01, respectively). In the PFO subgroup a negative correlation was found between the L-Arg/ADMA ratio and PFO tunnel length/height ratio (p ≤ 0.05; r = − 0.37; R2 = 0.14). Interestingly, the L-Arg/ADMA ratio varied across MTHFR genotypes (p ≤ 0.0001) and was lower in subgroups carrying the most impaired enzyme with respect to patients carrying the conservative MTHFR (p ≤ 0.0001, p ≤ 0.05, respectively). Consistently, OS patients carried the most dysfunctional MTHFR genotypes, whereas SI patients the least ones. Conclusions: A low L-Arg/ADMA ratio correlates with impaired activity of MTHFR and with the jeopardized IAS phenotype along a severity spectrum encompassing OS, PFO with long/tight tunnel, PFO with short/large tunnel, and SI. This infers that genetic MTHFR defects may underlie endothelial dysfunction-related IAS abnormalities, and predispose to a cryptogenic stroke. Our findings emphasize the role of the L-Arg/ADMA ratio as a reliable marker of stroke susceptibility in carriers of IAS abnormalities, and suggest its potential use both as a diagnostic tool and as a decision aid for therapy.

Association of MTHFR 677C > T genetic polymorphism with extrapyramidal side effects of antipsychotic treatment

Several studies have revealed a positive association between methylenetetrahydrofolate reductase (MTHFR) single nucleotide polymorphism 677C > T and schizophrenia. In T-allele carriers the functional activity of MTHFR is reduced contributing to hyperhomocysteinemia development, methionine deficiency and as a consequence - redox dysregulation and the alteration of dopamine synthesis. The aim of this study was to investigate the association of MTHFR677C > T with the severity of extrapyramidal side effects of antipsychotics in schizophrenia. The average score on the Simpson-Angus scale (SAS) in a group of patients with MTHFR 677 T-allele (n = 30; 13.27 ± 5.10, M ± SD) was statistically significantly higher (t = −2.40; p = 0.02) than in the comparison group of wild genotype MTHFR 677 CC carriers (n = 31; 9.84 ± 6.03, M ± SD), confirming the working hypothesis about a possible association of the MTHFR677 T-allele carriage with the development of extrapyramidal side effects of antipsychotics.

Riboflavin supplementation alters global and gene-specific DNA methylation in adults with the MTHFR 677 TT genotype

DNA methylation is important in regulating gene expression and genomic stability while aberrant DNA methylation is associated with disease. Riboflavin (FAD) is a cofactor for methylenetetrahydrofolate reductase (MTHFR), a critical enzyme in folate recycling, which generates methyl groups for homocysteine remethylation to methionine, the pre-cursor to the universal methyl donor S-adenosylmethionine (SAM). A polymorphism (C677T) in MTHFR results in decreased MTHFR activity and increased homocysteine concentration. Previous studies demonstrated that riboflavin modulates this phenotype in homozygous adults (MTHFR 677 TT genotype), however, DNA methylation was not considered. This study examined DNA methylation, globally and at key MTHFR regulatory sites, in adults stratified by MTHFR genotype and the effect of riboflavin supplementation on DNA methylation in individuals with the 677 TT genotype. Samples were accessed from participants, screened for the MTHFR C677T polymorphism, who participated in observational (n = 80) and targeted riboflavin (1.6 mg/day) RCTs (n = 80). DNA methylation at LINE-1 and key regulatory regions of the MTHFR locus were analysed by pyrosequencing in peripheral blood leukocytes. LINE-1 (+1.6%; p = 0.011) and MTHFR south shelf (+4.7%, p < 0.001) were significantly hypermethylated in individuals with the MTHFR 677 TT compared to CC genotype. Riboflavin supplementation resulted in decreased global methylation, albeit only significant at one CpG. A significant reduction in DNA methylation at the MTHFR north shore (−1.2%, p < 0.001) was also observed in TT adults following intervention with riboflavin. This provides the first RCT evidence that DNA methylation may be modulated by riboflavin in adults with the MTHFR 677 TT genotype.