Vitamin D Receptor Genotype Research Articles

A phase I study of high-dose calcitriol in combination with temozolomide for patients with metastatic melanoma.

Background: Temozolomide is efficacious as an oral alternative for patients with metastatic melanoma (MM). Calcitriol has anti-proliferative properties and vitamin D receptor (VDR) polymorphisms are associated with alterations in melanoma susceptibility and progression. Methods: Tem 150 mg/m2 was administered on days 2–8 and 16–22 every 28 days. Calcitriol was given on days 1 and 15 every 28 days. VDR gene analysis was completed using PCR-RFLP based assays. Tolerability was the primary objective with secondary objectives of time to progression (TTP) and overall survival (OS). Results: Twenty pts with MM were registered. Cytopenias and thrombosis were the most common grade 3 or 4 toxicities. Median TTP was 1.8 mo. Pts with high-risk VDR genotype tt+/−ff (n = 6) had an OS of 3.8 mo from time of enrollment, compared to 7.4 mo for those with non-tt/ff genotypes (n = 11), although not statistically significant (HR = 1.20, 95% CI 0.41–3.53, p = 0.74). Conclusions: The extended dosing of Tem with calcitriol is a well-tolerated regimen. The trend toward improved OS in non-tt/ff VDR genotypes is consistent with prior studies associating the tt/ff genotype with biologic aggressiveness.

Analysis on the genetic polymorphisms and haplotypes of vitamin D receptor in three hundred and eighty-two patients with ulcerative colitis

Objective To investigate the relationship between polymorphisms and haplotypes of vitamin D receptor (VDR) genes FokⅠ,BsmⅠ,ApaⅠ,TaqⅠ and ulcerative colitis (UC) in Han population of Zhejiang province.Methods From January 2004 to September 2012,382 patients with UC were collected and 489 age and gender matched healthy individuals were collected at the same period as controls.Two milliliter fasting peripheral blood of subjects was obtained for whole-genome DNA isolation.Microsequencing technology was applied.Polymnorphisms of VDR were determined through multiplex polymerase chain reaction (PCR) amplification,PCR products purification,extension,second purification and capillary electrophoresis genotype.Whether the distribution of VDR FokⅠ,BsmⅠ,ApaⅠ,Taq Ⅰ genotype of control group met Hardy-Weinberg equilibrium was analyzed by chi-square test.After age,gender and smoking factors adjusted,the differences of VDR FokⅠ,BsmⅠ,ApaⅠ,TaqⅠ allele and genotype frequencies between UC group and control group were analyzed by non-conditional Logistic regression analysis,and their effects on clinic-pathological features of patients with UC were also analyzed.Linkage disequilibrium and haplotype analysis was made by Haploview 4.2 software.Results The distributions of VDR FokⅠ,Bsm],ApaⅠ,TaqⅠ genotype met Hardy-Weinberg equilibrium (all P＞0.05).After age,gender and smoking factors adjusted,the results of non conditional Logistic regression analysis indicated that there were no significant differences in VDR FokⅠ,BsmⅠ,ApaⅠ,TaqⅠ allele and genotype frequencies between UC group and control group (all P＞0.05).However,the frequencies of mutant allele (C) and genotype (TC+CC) in VDR FokⅠ of patients with mild and moderate UC (54.14％,353/652 ; 81.60％,266/326) were significantly higher than those of patients with severe UC (39.29％,44/112; 57.14％,32/56; OR=1.825 and 3.322,95％CI 1.212 to 2.747 and 1.828 to 6.061,both P＜0.01).There was no correlation between VDR BsmⅠ,ApaⅠ,TaqⅠ and severity of UC (all P＞0.05).There were no significant differences in VDR FokⅠ,BsmⅠ,ApaⅠ,TaqⅠ allele and genotype frequencies between UC patients with distal colitis and extensive colitis (all P＞0.05).There were linkage disequilibrium in VDR BsmⅠ,ApaⅠ and TaqⅠ.Linkage disequilibrium coefficient (D') between BsmⅠ and ApaⅠ was 0.982 and the correlation coefficient (r2) was 0.156; D' between ApaⅠ and TaqⅠ was 0.929 and r2 was 0.141; D' between BsmⅠ and TaqⅠ was 0.826 and r2 was 0.675.Among eight haplotypes (GAC,ACC,GCC,AAT,ACT,GCT,GAT and AAC),the frequency of AAC haplotype of UC group (3.80％,29/764)was lower than that of control group (5.93％,58/978) and the difference was statistically significant (OR=0.631,95％CI 0.394-1.013,P=0.039).Conclusions The genetic mutation of VDR FokⅠ may influence severity of patients with UC.The risk of developing UC is low in the individuals carrying AAC haplotype of VDR BsmⅠ,ApaⅠ,TaqⅠ suffering UC. Key words: Colitis, ulcerative; Receptors, calcitriol; Gene polymorphism

Plasma vitamin D concentration influences survival outcome after a diagnosis of colorectal cancer.

We investigated whether the plasma level of 25-hydroxyvitamin D (25-OHD) after a diagnosis of colorectal cancer (CRC) influences survival outcome. We prospectively studied 1,598 patients with stage I to III CRC. We sought association between plasma 25-OHD and stage-specific survival and tested for interaction between 25-OHD level and variation at the vitamin D receptor (VDR) gene locus. Blood was sampled postoperatively, and plasma was assayed for 25-OHD by liquid chromatography-tandem mass spectrometry. VDR polymorphisms (rs1544410, rs10735810, rs7975232, rs11568820) were genotyped, and haplotypes were inferred by using BEAGLE software. We tested for association between survival and 25-OHD, VDR genotype/haplotype, and after applying a VDR genotype-25-OHD interaction term. We conducted Kaplan-Meier survival analysis and used Cox proportional hazards models to estimate adjusted hazard ratios (HRs). We found strong associations between plasma 25-OHD concentration and CRC-specific (P = .008) and all-cause mortality (P = .003). Adjusted HRs were 0.68 (95% CI, 0.50 to 0.90) and 0.70 (95% CI, 0.55 to 0.89), respectively (highest v lowest 25-OHD tertile), particularly in stage II disease (HR, 0.44; P = .004 for CRC-specific mortality). We detected gene-environment interactions between 25-OHD concentration and rs11568820 genotype for CRC-specific (P = .008) and all-cause (P = .022) mortality, number of protective alleles (P = .004 and P = .018, respectively), and GAGC haplotype at the VDR locus for all-cause mortality (P = .008). In patients with stage I to III CRC, postoperative plasma vitamin D is associated with clinically important differences in survival outcome, higher levels being associated with better outcome. We observed interactions between 25-OHD level and VDR genotype, suggesting a causal relationship between vitamin D and survival. The influence of vitamin D supplementation on CRC outcome will require further investigation.

Significance of Vitamin D Receptor Gene Polymorphisms for Risk of Hepatocellular Carcinoma in Chronic Hepatitis C

BACKGROUND/AIMSBiological and epidemiological data suggest that vitamin D levels may influence cancer development. Several single nucleotide polymorphisms have been described in the vitamin D receptor (VDR) gene in association with cancer risk. We aimed to investigate the association of VDR gene polymorphisms with hepatocellular carcinoma (HCC) development in chronic hepatitis C patients. METHODSIn a cross-sectional, hospital-based setting, 340 patients (201 chronic hepatitis, 47 cirrhosis and 92 HCC) and 100 healthy controls receiving VDR genotyping (bat-haplotype: BsmI rs1544410 C, ApaI rs7975232 C and TaqI rs731236 A) were enrolled. RESULTSPatients with HCC had a higher frequency of ApaI CC genotype (P = 0.027) and bAt[CCA]-haplotype (P = 0.037) as compared to control subjects. There were no differences in BsmI and TaqI polymorphisms between two groups. In patients with chronic hepatitis C, HCC subjects had a higher frequency of ApaI CC genotype and bAt[CCA]-haplotype than those with chronic hepatitis (P = 0.001 and 0.002, respectively) and cirrhosis (P = 0.019 and 0.026, respectively). After adjusting age and sex, logistic regression analysis showed that ApaI CC genotype (odds ratio: 3.02, 95% confident interval: 1.65-5.51) was independently associated with HCC development. CONCLUSIONVDR ApaI polymorphism plays a role in the development of HCC among chronic hepatitis C patients. Further explorations of this finding and its implications are required.

Melanoma risk is associated with vitamin D receptor gene polymorphisms

Previous studies have reported that vitamin D receptor (VDR) gene polymorphisms are associated with the occurrence of various cancers, including melanoma. The aim of the current study was to investigate the association of VDR gene polymorphisms with melanoma risk, clinicopathological characteristics, and vitamin D levels. The study group included 117 patients (84 patients with superficial spreading melanoma and 33 patients with nodular melanoma). The control group included 122 sex-matched and age-matched healthy-blood donors of the same ethnicity. VDR gene polymorphisms FokI, EcoRV, TaqI, and ApaI were genotyped by real-time PCR. In 60 patients, the total 25-hydroxyvitamin D levels were evaluated in serum samples by direct chemiluminescence. Associations among parameters were considered to be significant if the P value was less than 0.05. Significant differences in the frequencies of VDR genotypes were observed between cases and the control group for FokI and TaqI polymorphisms (P<0.0001; P=0.005, respectively). Heterozygous Ff as well as mutant FF genotypes of the FokI polymorphism were associated with increased melanoma risk compared with the wild-type form [odds ratio (OR)=3.035, P=0.003; OR=9.276, P<0.0001, respectively]. A significantly increased melanoma risk was observed for the heterozygous Tt (OR=2.302, P=0.011) and the mutated variant tt (OR=3.697, P=0.003) of the TaqI polymorphism in comparison with the wild-type genotype. None of the polymorphisms studied was associated with clinicopathological characteristics and vitamin D serum level. Our results suggest that FokI and TaqI polymorphisms in the VDR gene may be considered as potential biomarkers for melanoma susceptibility. Low vitamin D levels in melanoma patients indicate the need for vitamin D supplementation.

Genetic markers of treatment response to tumour necrosis factor-α inhibitors in the treatment of psoriasis.

Anti-tumour necrosis factor (TNF)-α therapies have revolutionized the treatment of psoriasis; however, up to 50% of patients do not respond satisfactorily. Identification of pharmacogenetic markers of treatment response is an important stop in the development of individually tailored treatment. The objective of this study was to assess the association of human leucocyte antigen (HLA)-C, killer immunoglobulin receptor (KIR) and vitamin D receptor (VDR) genotypes with response to treatment by etanercept and adalimumab. This was a study of 138 patients with severe chronic plaque psoriasis who were treated with etanercept and/or adalimumab. Patients were classified as responders if they achieved a 75% reduction in PASI (PASI75) or were almost clear of psoriasis after 24weeks of therapy. The frequencies of HLA-C and KIR haplotypes and VDR polymorphisms were compared in responders and nonresponders. The frequency of all HLA-C and KIR genotypes were compared between the 138 patients with psoriasis and 247 healthy donors. The number of patients classified as responders was 46 of 94 (49%) in the etanercept group and 50 of 76 (66%) in the adalimumab group. None of the HLA-C, KIR or VDR genotypes examined was predictive of treatment response. Compared with healthy controls, patients with psoriasis were more likely to have the HLA-C*06 genotype (P<0.001) and less likely to have the HLA-C*07 genotype (P<0.001), whereas there was no significant difference in frequencies of any KIR subtype. Using the candidate gene approach to identify biomarkers of treatment response in psoriasis may have limited utility. This was a small study with limited power. Future larger studies are needed to further examine these findings and to explore alternative approaches to identify predictors of treatment response to biological agents.

Vitamin D receptor gene polymorphism and 25 hydroxy vitamin D levels in Egyptian patients with pulmonary tuberculosis

BackgroundVitamin D is a potent immunomodulator of innate immune responses, and its deficiency was implicated in the activation of tuberculosis (TB). The effects of vitamin D are exerted by the interaction with the vitamin D receptor (VDR) and may be influenced by polymorphisms in the VDR gene. MethodsWe conducted a case–control study to identify 25(OH) D levels and FOK1 polymorphism of the VDR gene in a group of newly diagnosed pulmonary TB patients and compared to the findings in a group of healthy controls. ResultsObtained results revealed that 25(OH) D levels were significantly lower in the patients than in the controls (P<0.0001), and the prevalence of vitamin D insufficiency was 72.5% in TB patients. The frequencies of FokI genotypes determined from TB patients were 30% for FF , 50% for Ff and 20% for ff, while in controls, the frequency of FF, Ff and ff were present in a percentage of; 40%, 40% and 20%, respectively. There was no significant association between the genotype groups, TB patient and normal control, for FokI polymorphism (P>0.05). Also, no association between VDR genotype and 25(OH) D levels was found. ConclusionFokI polymorphisms in the VDR gene do not appear to be responsible for host susceptibility to human tuberculosis in the Egyptian population. However, vitamin D deficiency may be a cause of the susceptibility to tuberculosis.

Vitamin D receptor polymorphism FokI is associated with spontaneous idiopathic preterm birth in an Israeli population

ObjectiveThe active form of vitamin D (1,25[OH]2D3) has been established to have potent anti-proliferative, immuno-modulatory, and anti-microbial action in addition to its effects on bone. The nuclear vitamin D receptor (VDR) is expressed in the placenta-decidua, regulating genes associated with implantation and implantation immuno-tolerance. If VDR polymorphisms regulate VDR functionality at the placenta-decidua interface, VDR genotypes may be involved in idiopathic preterm birth (PTB). Study designMaternal and fetal (umbilical cord) blood samples from 33 Jewish and Arab mothers with PTB of a singleton neonate were compared to 98 samples from Jewish and Arab maternal and fetal blood samples from full-term, uncomplicated singleton births. Maternal age and ethnicity were comparable between groups. PCR amplification/digestion identified the VDR SNPs: FokI, ApaI, TaqI, and BsmI. ResultsAllele frequency for the FokI VDR in maternal blood samples from preterm births (but not umbilical cord samples) was significantly different (p=0.01) from that in maternal and umbilical cord blood samples from full-term singleton births, with an odds ratio for FokI carriers of 3.317 (95% CI, 1.143, 9.627) for preterm birth. The FokI VDR variant may therefore be a maternal risk trait for PTB among these women. ConclusionThis study may support a future platform for the study of vitamin D during pregnancy and treatment of selective target populations with vitamin D and/or VDR “tissue-specific therapeutic intervention” for prevention of PTB.

Vitamin D Receptor Gene Polymorphism and Left Ventricular Hypertrophy in Chronic Kidney Disease

FokI and BsmI polymorphisms of vitamin D receptor (VDR) gene are regarded as reliable markers of disturbed vitamin D signaling pathway. Left ventricular hypertrophy (LVH) is a strong cardiovascular risk marker in end stage renal disease (ESRD) patients. Since BsmI polymorphism has been associated with LVH in ESRD patients, we addressed this study in patients with chronic kidney disease (CKD) not yet on dialysis. One hundred and forty five patients with CKD stage 3 were genotyped for FokI and BsmI VDR polymorphisms, in order to assess the relationships between these VDR polymorphisms, some markers of mineral bone disorders, and LVH measured by echocardiography. Patients bearing either the Ff heterozygous or FF homozygous genotype had significantly higher PTH values than those bearing the ff genotype. The relationships between VDR genotypes and LVH revealed a highly significant association of the BsmI Bb heterozygous genotype with LVH. In patients with CKD stage 3 BsmI B allele was independently related to LVH. Since LVH is a frequent finding in dialysis population due to several mechanisms, the presence of the same relationship in patients with CKD strengthens the hypothesis that alterations of vitamin D signaling are implicated in LVH development in patients with renal diseases.

Vitamin D Receptor Genetics on Extracellular Matrix Biomarkers and Hemodynamics in Systolic Heart Failure

Vitamin D deficiency has been associated with the development of myocardial hypertrophy and inflammation. These findings suggest that vitamin D status and vitamin D receptor (VDR) genomics may play a role in myocardial fibrosis. The aim of this pilot study was to determine the association between vitamin D levels, VDR polymorphisms, and biomarkers of left ventricular remodeling and hemodynamics. In a cross-sectional pilot study, patients with ejection fraction (EF) <40% (and New York Heart Association ≥ II) undergoing right heart catheterization were included in the study. Blood was collected for determination of 25-hydroxyvitamin D level (antibody competitive immunoassay), VDR genotypes (BsmI, ApaI, TaqI, and FokI), and biomarkers (N-terminal propeptide of collagen type III [PIIINP], matrix metalloproteinase 2, and galectin 3). The vitamin D genotypes were determined through the use of pyrosequencing. A total of 30 patients with a mean EF of 17% ± 8% were enrolled. There was a significant association between the BsmI C allele, ApaI G allele, and TaqI A allele, which formed a haplotype block (CGA) for analysis. There were no differences in baseline parameters between patients with the VDR haplotype block (n = 20) and those without (n = 10). Individual genotypes were not associated with any biomarker or hemodynamics. Patients with the CGA haplotype demonstrated significantly higher log PIIINP values (1.74 ± 0.32 mcg/mL vs 1.36 ± 0.31 mcg/mL, P = .0041). When evaluating vitamin D levels below and above the median level (19 ng/mL), there was no significant difference between these 2 groups in regard to biomarker levels for left ventricular remodeling. This study has shown that a biomarker for collagen type III synthesis, PIIINP, was associated with the CGA haplotype of BsmI, ApaI, and TaqI single nucleotide polymorphisms on the VDR. These findings suggest that VDR genetics may play a role in myocardial fibrosis in patients with systolic heart failure.

Juvenile idiopathic arthritis patients and their skeletal status: possible role of vitamin D receptor gene polymorphism

We evaluated bone mineralization and metabolism changes related to vitamin D receptor (VDR) polymorphic genotypes in children with juvenile idiopathic arthritis. One hundred and ninety eight children (82 boys and 116 girls) were included in our study. Bone mineral density (BMD) was measured by lumbar spine DXA. Osteocalcin, CTX, parathyroid hormone, total and ionized calcium, inorganic phosphate, total alkaline phosphatase activity was utilized for assessment of bone metabolism. Molecular testing: TaqI (rs731236) and Cdx2 (rs11568820) polymorphisms of VDR were detected by RFLP. No differences in TaqI and Cdx2 haplotypes, genotypes and alleles distribution related with normal and low BMD (Zscore <-2SD) were found. Children with low linear growth (<10th percentile) had more allele T-contained genotypes of TagI VDR (p = 0.037), compare with medium or high linear growth children. Children with high linear growth (>90th percentile) had the highest frequency of allele A-contained genotypes (GA+AA) of Cdx2 VDR (p = 0.009). Girls with TT TaqI VDR, who never been treated by glucocorticoides had lower BMD-Zscore than C allele carriers (TT = -0.94SD [IQR: -2.1;-0.5], TC+CC = -0.62SD [IQR: -1.26;0.39], p = 0.03). Girls with Tanner I with TT had higher total and ionized Ca level than carriers of C allele (Ca: TT = 2.43 ± 0.15 mmol/l, TC+CC = 2.28 ± 0.2 mmol/l, p = 0.024; Ca(2+): TT = 1.15 ± 0.08 mmol/l, TC+CC = 1.06 ± 0.13 mmol/l, p = 0.026). Presence of TT genotype negatively correlated with BMD-Zscore (r = -0.28, p = 0.04), and positively with frequency of LBMD (r = 0.3, p = 0.037). Boy with GG Cdx2 genotype had lower total Ca (GG = 2.3 ± 0.17 mmol/l, GA+AA = 2.43 ± 0.17 mmol/l, p = 0.004) compare with carriers of A allele. Pubertal boys (Tanner IV-V) with GG had higher CTX (GG = 1.75 ± 0.11 ng/ml, GA+AA = 1.06 ± 0.07 ng/ml, p = 0.04. TT genotype of TaqI and GG genotype of Cdx2 VDR is a negative factor impact bone mineralization metabolism and linear growth.

The role of vitamin D deficiency and vitamin d receptor genotypes on the degree of collateralization in patients with suspected coronary artery disease.

We determined the association of vitamin D deficiency and the FokI polymorphism of the vitamin D receptor (VDR) gene in 760 patients who underwent angiography due to suspected coronary artery disease (CAD). Angiography and the Rentrop scoring system were used to classify the severity of CAD in each patient and to grade the extent of collateral development, respectively. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to determine the FokI VDR gene polymorphism. The prevalence of severe vitamin D deficiency (serum 25(OH)D < 10 ng/mL) was significantly higher in patients with at least one stenotic coronary artery compared to those without any stenotic coronary arteries. Severe vitamin D deficiency was not independently associated with collateralization, but it was significantly associated with the VDR genotypes. In turn, VDR genotype was independently associated with the degree of collateralization; the Rentrop scores were the highest in FF, intermediate in Ff, and the lowest in the ff genotype. The results show that FokI polymorphism is independently associated with collateralization. Additionally, vitamin D deficiency is more prevalent in patients with CAD that may result from FokI polymorphism. Therefore, maintaining a normal vitamin D status should be a high priority for patients with CAD.

Vitamin D Receptor Genotype Modulates the Correlation between Vitamin D and Circulating Levels of let-7a/b and Vitamin D Intake in an Elderly Cohort

Background and Aims: Circulating microRNAs (miRNAs) are linked to disease and are potential biomarkers. Vitamin D may modulate miRNA profiles, and vitamin D status has been linked to risk of disease, including cardiovascular disease and cancers. We hypothesise that genotypic variance influences these relationships. We examined the correlations between vitamin D intake and circulating levels of the miRNAs let-7a/b, and the involvement of two common vitamin D receptor (VDR) polymorphisms, BsmI and ApaI. Methods: Two hundred participants completed food frequency and supplement questionnaires, and were assayed for circulating let-7b expression by qPCR. Polymorphisms were detected using restriction fragment length polymorphism-PCR. Results: let-7b expression negatively correlated with vitamin D intake (r_s = -0.20, p = 0.005). The magnitude and direction of correlation were maintained in the presence of the BsmI restriction site (r_s = -0.27, p = 0.0005). However, in the absence of BsmI restriction site, the direction of the correlation was reversed (r_s = +0.319, p = 0.0497). These correlations were significantly different (z-score = 2.64, p = 0.0085). The correlation between vitamin D intake and let-7a was only significant in those without the ApaI restriction site. Conclusions: The correlation between vitamin D intake and let-7a/b expression in this cohort varies with VDR genotype. This study highlights the importance of considering underlying genotypic variance in miRNA expression studies and in nutritional epigenetics generally.

Vitamin D receptor polymorphisms and cancer.

It was suggested that vitamin D levels influence cancer development. The vitamin D receptor (VDR) is a crucial mediator for the cellular effects of vitamin D. In fact It has been hypothesized that polymorphisms in the VDR gene affect cancer risk and the relevance of VDR gene restriction fragment length polymorphisms for various types of cancer has been investigated by a great number of studies. However, results from previous studies on the association of VDR polymorphisms with different cancer types are somewhat contradictory, and the role of VDR in the etiology of cancer is still equivocal. We have performed a systematic review of the literature to analyze the relevance of more VDR polymorphisms (Fok1, Bsm1, Taq1, Apa1, and Cdx2) for individual malignancies, including cancer of the skin (melanoma and nonmelanoma skin cancer), ovarian cancer, renal cell carcinoma, bladder cancer, non-Hodgkin lymphoma, leukemia, thyroid carcinoma, esophageal adenocarcinoma, hepatocellular carcinoma, sarcoma, head and neck and oral squamous cell carcinoma. Up to June 2012, we identified 79 independent studies for a total of 52427 cases and 62225 controls. Significant associations with VDR polymorphisms have been reported for prostate (Fok1, Bsm1, Taq1), breast (Fok1, Bsm1, Apa1), colon-rectum (Fok1, Bsm1, Taq1) and skin cancer (Fok1, Bsm1, Taq1). Very few studies reported risk estimates for the other cancer sites. Conflicting data have been reported for most malignancies and at present it is still not possible to make any definitive statements about the importance of the VDR genotype for cancer risk. It seems probable that interactions with other factors such as calcium and vitamin D intake, 25(OH)D plasma levels and UV radiation exposure play a decisive role in cancer risk. To conclude, there is some indication that VDR polymorphisms may modulate the risk of some cancer sites and in future studies VDR genetic variation should be integrated also with prediagnostic indicator of vitamin D status.

Rapid and Simultaneous Detection of Vitamin D Receptor Gene Polymorphisms by a Single ARMS-PCR Assay

Vitamin D has various roles in many biological actions such as calcium homeostasis, cell proliferation, and cell differentiation to many target tissues. These effects are mediated by the active form of vitamin D, 1,25(OH)2D3, which binds to a cytoplasmic protein called vitamin D receptor (VDR). VDR gene has four common single nucleotide polymorphisms (SNPs) that are defined by the presence of restriction sites for FokI (F/f), TaqI (T/t), BsmI (B/b), and ApaI (A/a). The association of VDR gene polymorphisms with several diseases has been investigated. In most studies, VDR genotyping was performed by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) assays, which are cumbersome and time consuming, and their results are sometimes difficult to interpret. We modified previously reported primers for VDR genotyping and set up a single amplification-refractory mutation system (ARMS)-PCR method for simultaneous genotyping of four common VDR polymorphisms. In this study, 218 DNA samples were analyzed for VDR genetic variants by this ARMS-PCR technique; 136 of them were re-genotyped by PCR-RFLP assays to compare genotyping results. We obtained allelic frequencies of 69 vs. 31% for F/f, 34 vs. 66% for B/b, 70 vs. 30% for T/t, and 52 vs. 48% for A/a in this sample of the Iranian population. In addition, comparisons of the results of these two methods showed good uniformity in VDR genotypes; although, in some samples, ambiguity in restriction patterns was present. As ARMS-PCR is more rapid, economic, and user friendly than PCR-RFLP, its substitution would be welcomed in disease association and pharmacogenetic studies of VDR variants.

Association between Vitamin D receptor (VDR) polymorphisms and matrix metalloproteinase-9 (MMP-9) in female workers of lead battery plants

Background Vitamin D receptor (VDR) genotype is a risk factor for several disorders with inflammatory components, including coronary heart diseases (CHD). Matrix metalloproteinases (MMPs), a group of zinc-dependent endopeptidases, involve extracellular matrix (ECM) physiological degradation process and contribute to progressive left ventricular remodeling, dilation, and heart failure. Aims To investigate whether the VDR genetic polymorphisms are associated with the activity of MMP-9 in female workers of lead battery plants. Methods A total of 206 female workers was recruited for this study at the Taiwan battery manufacturing plant. The mean age was 46.7 ± 8.5 years and mean working duration was 12.8 ± 8.1 years. A clinical examination was performed at a workplace health check-up, and information was obtained using self-reported questionnaire. Each study participant provided blood samples for genotyping of VDR (folk, bsml, apal) and for serum MMP-9, whole blood lead levels (BLLs) and serum oxidative stress (Malondialdehyde (MDA)) measurements. We used multiple linear regression models for determinants of MMP-9 using VDR genotypes, BLLs and MDA as independent variables Results The MMP-9 levels was found to be correlated with BLLs and menopausal status in female workers. In the trend analysis, results exhibited a significant decreased trend in MMP-9 levels from the wild-type, heterogeneity and homozygousgenotypes of VDR (apal and bsml). After adjusting for determinants of MMP-9 levels,we found that carriers of the AA genotype of VDR (apal) had a lower MMP-9 levelcompared with carriers of CC genotype in these subjects (?=-0.651, p=0. 012). A similar result was observed in carriers of the AG genotype of VDR (bsml) compared with carriers of CC genotype (?=-0.641, p= 0.008). Conclusions This study confirms that the BLLs and menopausal status are important determinants in the MMP-9 levels. There is a possibility that female workers who carry homozygous genotypes of VDR (apal and bsml) decrease in MMP-9 levels.

SAT0467 Long Term Outcome and Foki VDR Polimorphism in JIA Patients on Anti-TNF Treatment

BackgroundContribution of FoxI VDR (vitamin D receptor) polymorphism on disease outcome in JIA is not yet well established.ObjectivesTo investigate influence of FoxI VDR polymorphism on long term outcome in JIA...

A phase I study of high-dose calcitriol in combination with temozolomide for patients (Pts) with metastatic melanoma (MM).

9087 Background: Temozolomide (Tem) has demonstrated efficacy as an oral alternative for pts with MM on traditional and extended dosing schedules. Calcitriol has known antiproliferative properties in vitro, and has shown synergistic effects with chemotherapy. Additionally, vitamin D receptor (VDR) polymorphisms are associated with alterations in melanoma susceptibility and disease progression. Specifically, the VDR genotype tt/ff (Taq1 and Fok1 polymorphisms) has been associated with tumors &gt;3.5mm, though no studies have focused on survival. Methods: Tem 150mg/m2was administered on days 2-8 and 16-22 every 28 days until progression or significant toxicity. Calcitriol was given on days 1 and 15 every 28 days with 3 pts at a dose of 0.2mcg/kg, 3 pts at 0.3mcg/kg, 3 pts at 0.5mcg/kg, and an additional 11 pts at a maximum dose of 0.5mcg/kg. VDR gene analysis was completed on 17/20 pts using PCR-RFLP based assays. Tolerability was the primary objective with secondary objectives of time to progression (PFS) and overall survival (OS), both as a whole and by VDR genotype. Results: Twenty pts (males=15) with MM treated with at least one prior systemic therapy or who were not candidates for interleukin-2 (conducted pre BRAF and anti CTLA agents), were accrued. Median age was 58. The regimen was well-tolerated with leukopenia, lymphopenia, thrombocytopenia, anemia, and thrombosis as the most common grade 3 or 4 toxicities at 10% incidence each, less than observed in prior studies (Patel et al Eur J Ca 2011). Obj RR was 10%. Median PFS was 1.8 mo with a mean of 2.7 cycles given. Pts with low-risk VDR genotype non-tt or ff (n= 11) had a median OS of 7.4 mo compared to 3.8 mo for tt or ff or both (n=6)(median ratio=1.95) from time of enrollment, although not statistically significant given small sample sizes. Conclusions: The extended dosing of Tem with calcitriol is a well-tolerated oral regimen in MM. The trend toward improved OS in non-tt/ff VDR genotypes is consistent with prior studies associating the tt/ff genotype with biologic aggressiveness. Whether this represents a benefit of the inclusion of calcitriol should be studied further. Clinical trial information: NCT00301067.

Association of vitamin D receptor gene polymorphisms with pancreatic cancer: A pilot study in a North China Population

Polymorphisms of the vitamin D receptor (VDR) gene may be a risk factor for pancreatic cancer (PC). We investigated the association of two single-nucleotide polymorphisms (SNPs) of the VDR gene with PC in age- and gender-matched patients and controls. PC (n=91) and healthy control (n=80) samples were genotyped for the FokI (rs2228570) and BsmI (rs1544410) polymorphisms using the PCR and restriction fragment length polymorphism (PCR-RFLP) method. Chi-square analysis was used to test for the overall association of VDR genotype with disease. There was a significant difference in the frequency of genotype FF between the PC patients and controls (Ptrend=0.009); however, the difference in frequency of genotype BB between the two groups was not significant (Ptrend=0.082). The difference between FF and Ff/ff frequency was significant (P=0.002). The two high-risk genotypes were ffbb and Ffbb, with an 11.66- and 6.42-fold increased risk of PC, respectively. VDR gene polymorphisms were important for the development of PC in this study population; however, further exploration of these findings and their implications are required.

Highlights of This Issue

Although alcohol consumption is an established risk factor for incident breast cancer, its role in breast cancer prognosis is unclear. Kwan and colleagues evaluated postdiagnosis alcohol consumption with both cancer recurrence and mortality among 9,329 breast cancer patients in the After Breast Cancer Pooling Project. The authors report that regular alcohol intake was not associated with risk of breast cancer recurrence; however, postmenopausal women who regularly consumed alcohol had increased risk of recurrence.There is a positive association between calcium intake and prostate cancer risk. Rowland and colleagues evaluated whether this association was modified by particular vitamin D receptor (VDR) genotypes that influence calcium absorption. Among men with a low calcium absorption VDR genotype, those in the lowest quartile of calcium intake had a 50% reduction in prostate cancer risk compared with those in the upper quartiles of calcium intake. These findings are consistent with the hypothesis that genetic determinants of calcium absorption influence prostate cancer risk and may contribute to racial disparities in prostate cancer incidence and mortality rates.The interpretation of uterine cancer rates is complicated because risk estimates often include women who have already undergone hysterectomies. To address this, Siegel and colleagues used population-based cancer registries, U.S. Census data and data from the Behavioral Risk Factor Surveillance System to estimate hysterectomy-corrected, age-adjusted uterine cancer rates. The authors report that stateby-state hysterectomy-corrected uterine cancer incidence rates were consistently higher than uncorrected rates. Failure to adjust uterine cancer rates for hysterectomy prevalence distorts the true geographic and racial patterns.There is strong association between high viral load (VL) of human papillomavirus (HPV) type 16 and risk for cervical cancer in situ (CIS), yet little data is available regarding HPV16 and invasive SCC. Sundström and colleagues evaluated HPV16 VL in women with CIS and invasive SCC and found low VLs were common among both CIS and SCC cases. In SCC cases, high HRV16 VL conferred increased risk, but only from 3 years before diagnosis, which differs considerably from CIS cases. HPV16 VL dynamics are complex in SCC, which may limit its use in screening.