VEGF Single Nucleotide Polymorphisms Research Articles

Hypoxia-Inducible Pathway Polymorphisms and Their Role in the Complications of Prematurity.

Excessive oxidative stress resulting from hyperoxia or hypoxia is a recognized risk factor for diseases of prematurity. However, the role of the hypoxia-related pathway in the development of these diseases has not been well studied. Therefore, this study aimed to investigate the association between four functional single nucleotide polymorphisms (SNPs) in the hypoxia-related pathway, and the development of complications of prematurity in relation to perinatal hypoxia. A total of 334 newborns born before or on the 32nd week of gestation were included in the study. The SNPs studied were HIF1A rs11549465 and rs11549467, VEGFA rs2010963, and rs833061. The findings suggest that the HIF1A rs11549465T allele is an independent protective factor against necrotizing enterocolitis (NEC), but may increase the risk of diffuse white matter injury (DWMI) in newborns exposed to hypoxia at birth and long-term oxygen supplementation. In addition, the rs11549467A allele was found to be an independent protective factor against respiratory distress syndrome (RDS). No significant associations with VEGFA SNPs were observed. These findings indicate the potential involvement of the hypoxia-inducible pathway in the pathogenesis of complications of prematurity. Studies with larger sample sizes are needed to confirm these results and explore their clinical implications.

Association of VEGF cytokine levels and single nucleotide polymorphisms of the <i>VEGF-A</i> gene with the genital endometriosis in the female population of the Northwestern Federal District of Russia

Aim. To study the association of neoangiogenesis VEGF-A gene polymorphisms C(-460)T (rs833061), C(+936)T (rs3025039) with the risk of genital endometriosis in the population of the Northwestern Federal District; to examine the relationship of polymorphic variants of the VEGF-A neoangiogenesis gene C(-460)T (rs833061), C(+936)T (rs3025039) with the concentration of VEGF-A factor in the blood of females with genital endometriosis.
 Materials and methods. Eighty-five female volunteers aged from 19 to 44 (mean age 32.9 (5.8) years) with a histologically confirmed diagnosis of genital endometriosis were included in the stud; also, 79 females without endometriosis according to diagnostic laparoscopy for infertility or ectopic pregnancy, aged 20 to 42 (mean age 32.5 (7.2) years, p=0.71) were included in the study. The obtained data were analyzed using SPSS 20.0 statistical software package; 2 value and p value were estimated.
 Results. In our study, we found an association between the C allele and the CC genotype of the C(+936)T (rs3025039) polymorphism of the VEGF-A gene with genital endometriosis: odds ratio (OR) 2.35, 95% confidence interval (CI) 1.034.61, p=0.023; OR 1.89, 95% CI 1.034.61, p=0.048, respectively. The TT genotype of the C(-460)T (rs833061) polymorphisms and the TT genotype and the C(+936)T (rs3025039) allele of the VEGF-A gene were less common in patients with genital endometriosis: OR 0.43, 95% CI 0.141.29, p=0.023; OR 0.06, 95% CI 0.043.18, p=0.001; OR 0.43, 95% CI 0.231.29, p=0.023 respectively. The blood concentration of VEGF-A cytokine was lower in patients with genital endometriosis at all genotypes of C(-460)T (rs833061) locus of VEGF-A gene than that in females without endometriosis. The analysis of genotype frequency distribution of polymorphic site C(+936)T of the VEGF-A gene showed that the VEGF-A cytokine level was 1.5 times higher in the comparison group with heterozygous ST genotype versus that in homozygous genotypes. Enzyme immunoassay in the examined females showed a 2.5-fold decrease in the level of the VEGF-A angiogenic cytokine in blood serum in patients with endometriosis compared to those without endometriosis.
 Conclusion. To better understand the pathogenetic features of the endometriosis course and to make an individual prognosis of the course and therapy efficacy, an additional analysis of the associations between the polymorphisms of the listed genes and the angiogenesis factor level is required.

VEGF single nucleotide polymorphisms predict improved outcome in advanced non-small cell lung cancer patients treated with platinum-based chemotherapy

We aimed to investigate the prognostic role of genetic variants of VEGF in advanced NSCLC patients treated with platinum-based chemotherapy. A total of 196 patients with advanced NSCLC treated with first-line platinum-based chemotherapy were enrolled. We evaluated the relationship between VEGF polymorphisms and efficacy outcomes and chemotherapy toxicity. We found that rs699947, rs833061 and rs1005230 were in full linkage disequilibrium. Patients with CC genotype of rs833061 had a significant longer PFS than TT genotype (CC vs TT, HR = 1.67, 95%CI = 1.01−2.76, P = 0.043). Patients harbouring CC genotype had longer PFS compared with CT genotype (P < 0.001). Moreover, CC genotypes conferred a significantly increased PFS compared to CT and TT genotype in dominant model (CC vs CT + TT, HR = 1.95, 95%CI = 1.23−3.10, P = 0.005). Patients carrying TT genotype of rs833061 had improved both ORR (HR = 0.54, 95%CI = 0.30−0.98, P = 0.041) and DCR (HR = 0.37, 95%CI = 0.20−0.66, P = 0.001) than non-TT patients. Furthermore, no association was found between any rs833061 alleles and adverse events (P = 0.425), but patients carrying rs1570360 AA genotype were more likely to experience grade 3−4 toxicities (P = 0.004) (GG vs AA, HR = 3.16, 95%CI = 1.26−7.94, P = 0.015). In conclusion, the variant homozygote CC of rs833061 exhibited a better prognosis based on association analysis. The present study provides reference for the future study of platinum-based chemotherapy response and toxicity.

Influence of polymorphisms in the vascular endothelial growth factor gene on allograft rejection after kidney transplantation: a meta-analysis

Background: Reported associations of allograft rejection in kidney transplant patients with VEGF single nucleotide polymorphisms (SNPs) have been inconsistent between studies, which prompted a meta-analysis to obtain more precise estimates. Methods: Using the PICO elements, kidney transplant patients (P) were compared by genotype data between rejectors (I) and non-rejectors (C) in order to determine the risk of allograft rejection (O) attributed to the VEGF SNPs. Literature search of four databases yielded seven articles. To calculate risks for allograft rejection, four SNPs were examined. Using the allele-genotype model we compared the variant ( var) with the wild-type ( wt) and heterozygous ( var- wt) alleles. Meta-analysis treatments included outlier and subgroup analyses, the latter was based on ethnicity (Indians/Caucasians) and rejection type (acute/chronic). Multiple comparisons were corrected with the Bonferroni test. Results: Five highly significant outcomes (P a < 0.01) survived Bonferroni correction, one of which showed reduced risk for the var allele (OR 0.61, 95% CI 0.45-0.82). The remaining four indicated increased risk for the wt allele where the chronic rejection (OR 2.10, 95% CI 1.36-3.24) and Indian (OR 1.44, 95% CI 1.13-1.84) subgroups were accorded susceptibility status. Conclusions: Risk associations for renal allograft rejection were increased and reduced on account of the wt and var alleles, respectively. These findings could render the VEGF polymorphisms useful in the clinical genetics of kidney transplantation.

VEGF-A, VEGFR1 and VEGFR2 single nucleotide polymorphisms and outcomes from the AGITG MAX trial of capecitabine, bevacizumab and mitomycin C in metastatic colorectal cancer

The phase III MAX clinical trial randomised patients with metastatic colorectal cancer (mCRC) to receive first-line capecitabine chemotherapy alone or in combination with the anti-VEGF-A antibody bevacizumab (± mitomycin C). We utilised this cohort to examine whether single nucleotide polymorphisms (SNPs) in VEGF-A, VEGFR1, and VEGFR2 are predictive of efficacy outcomes with bevacizumab or the development of hypertension. Genomic DNA extracted from archival FFPE tissue for 325 patients (69% of the MAX trial population) was used to genotype 16 candidate SNPs in VEGF-A, VEGFR1, and VEGFR2, which were analysed for associations with efficacy outcomes and hypertension. The VEGF-A rs25648 ‘CC’ genotype was prognostic for improved PFS (HR 0.65, 95% CI 0.49 to 0.85; P = 0.002) and OS (HR 0.70, 95% CI 0.52 to 0.94; P = 0.019). The VEGF-A rs699947 ‘AA’ genotype was prognostic for shorter PFS (HR 1.32, 95% CI 1.002 to 1.74; P = 0.048). None of the analysed SNPs were predictive of bevacizumab efficacy outcomes. VEGFR2 rs11133360 ‘TT’ was associated with a lower risk of grade ≥ 3 hypertension (P = 0.028). SNPs in VEGF-A, VEGFR1 and VEGFR2 did not predict bevacizumab benefit. However, VEGF-A rs25648 and rs699947 were identified as novel prognostic biomarkers and VEGFR2 rs11133360 was associated with less grade ≥ 3 hypertension.

Influence of polymorphisms in the vascular endothelial growth factor gene on allograft rejection after kidney transplantation: a meta-analysis.

Background: Reported associations of allograft rejection in kidney transplant patients with VEGF single nucleotide polymorphisms (SNPs) have been inconsistent between studies, which prompted a meta-analysis to obtain more precise estimates. Methods: Using the PICO elements, kidney transplant patients (P) were compared by genotype data between rejectors (I) and non-rejectors (C) in order to determine the risk of allograft rejection (O) attributed to the VEGF SNPs. Literature search of four databases yielded seven articles. To calculate risks for allograft rejection, four SNPs were examined.Using the allele-genotype model we compared the variant ( var) with the wild-type ( wt) and heterozygous ( var- wt) alleles. Meta-analysis treatments included outlier and subgroup analyses, the latter was based on ethnicity (Indians/Caucasians) and rejection type (acute/chronic). Multiple comparisons were corrected with the Bonferroni test. Results: Five highly significant outcomes (P a < 0.01) survived Bonferroni correction, one of which showed reduced risk for the var allele (OR 0.61, 95% CI 0.45-0.82). The remaining four indicated increased risk for the wt allele where the chronic rejection (OR 2.10, 95% CI 1.36-3.24) and Indian (OR 1.44, 95% CI 1.13-1.84) subgroups were accorded susceptibility status. Conclusions: Risk associations for renal allograft rejection were increased and reduced on account of the wt and var alleles, respectively. These findings could render the VEGF polymorphisms useful in the clinical genetics of kidney transplantation.

Phase II trial of carboplatin and bevacizumab in patients with breast cancer brain metastases

BackgroundWe aimed to examine the safety and efficacy of bevacizumab and carboplatin in patients with breast cancer brain metastases.MethodsWe enrolled patients with breast cancer and > 1 measurable new or progressive brain metastasis. Patients received bevacizumab 15 mg/kg intravenously (IV) on cycle 1 day 1 and carboplatin IV AUC = 5 on cycle 1 day 8. Patients with HER2-positive disease also received trastuzumab. In subsequent cycles, all drugs were administered on day 1 of each cycle. Contrast-enhanced brain MRI was performed at baseline, 24–96 h after the first bevacizumab dose (day + 1), and every 2 cycles. The primary endpoint was objective response rate in the central nervous system (CNS ORR) by composite criteria. Associations between germline VEGF single nucleotide polymorphisms (rs699947, rs2019063, rs1570360, rs833061) and progression-free survival (PFS) and overall survival (OS) were explored, as were associations between early (day + 1) MRI changes and outcomes.ResultsThirty-eight patients were enrolled (29 HER2-positive, 9 HER2-negative); all were evaluable for response. The CNS ORR was 63% (95% CI, 46–78). Median PFS was 5.62 months and median OS was 14.10 months. As compared with an Eastern Cooperative Oncology Group performance status (ECOG PS) of 0, patients with ECOG PS 1–2 had significantly worse PFS and OS (all P < 0.01). No significant associations between VEGF genotypes or early MRI changes and clinical outcomes were observed.ConclusionsThe combination of bevacizumab and carboplatin results in a high rate of durable objective response in patients with brain metastases from breast cancer. This regimen warrants further investigation.Trial registrationNCT01004172. Registered 28 October 2009.

VEGF-Related Germinal Polymorphisms May Identify a Subgroup of Breast Cancer Patients with Favorable Outcome under Bevacizumab-Based Therapy-A Message from COMET, a French Unicancer Multicentric Study.

The prospective multicenter COMET trial followed a cohort of 306 consecutive metastatic breast cancer patients receiving bevacizumab and paclitaxel as first-line chemotherapy. This study was intended to identify and validate reliable biomarkers to better predict bevacizumab treatment outcomes and allow for a more personalized use of this antiangiogenic agent. To that end, we aimed to establish risk scores for survival prognosis dichotomization based on classic clinico-pathological criteria combined or not with single nucleotide polymorphisms (SNPs). The genomic DNA of 306 patients was extracted and a panel of 13 SNPs, covering seven genes previously documented to be potentially involved in drug response, were analyzed by means of high-throughput genotyping. In receiver operating characteristic (ROC) analyses, the hazard model based on a triple-negative cancer phenotype variable, combined with specific SNPs in VEGFA (rs833061), VEGFR1 (rs9582036) and VEGFR2 (rs1870377), had the highest predictive value. The overall survival hazard ratio of patients assigned to the poor prognosis group based on this model was 3.21 (95% CI (2.33–4.42); p < 0.001). We propose that combining this pharmacogenetic approach with classical clinico-pathological characteristics could markedly improve clinical decision-making for breast cancer patients receiving bevacizumab-based therapy.

Cervical transverse MRI in ALS diagnosis and possible link to VEGF and MMP9 single nucleotide polymorphisms. Case Report

We report the case of a patient diagnosed with ALS and presenting with a “snake eyes” sign on axial cervical MRI. Given the peculiar radiological presentation and our patient’s young age, we decided to perform a genetic analysis to better investigate the molecular mechanisms underlying the pathogenesis of his clinical phenotype. In our study, we identified different polymorphisms of interest at the level of VEGF and MMP9 promoters that we suggest could contribute to neuronal sufferance by promoting vascular abnormalities and tissue hypoxia, in accordance with previous published papers. When combined with repetitive mechanical stress at cervical level, as evident in our patient’s case, the resultant radiological image appears to be the “snake eyes” sign in the anterior horns of the cervical spinal cord, while the clinical phenotype is that of an atypical ALS case, possibly an ALS-mimic. To correctly identify and diagnose such patients, in particular when disease course and neuroradiological data do not correspond to those of a classical ALS case, we underline the importance of axial cervical MRI studies and suggest the possibility of their implementation into routine clinical practice.

Angiogenesis Genotyping and Clinical Outcomes in Patients with Advanced Hepatocellular Carcinoma Receiving Sorafenib: The ALICE-2 Study.

Sorafenib represents one of the therapeutic strongholds for advanced hepatocellular carcinoma (HCC), but unfortunately, predictive factors are lacking. We previously reported that the VEGF single nucleotide polymorphisms (SNPs) rs2010963 and rs4604006 might correlate with clinical outcomes in sorafenib-treated HCC patients. The objective of the ALICE-2 study is to define a prognostic angiogenesis profile to better identify HCC patients who are more likely to benefit from sorafenib treatment. From 2008 to 2015, all consecutive HCC patients receiving sorafenib according to the Italian label were tested for specific HIF-1α, VEGF, and VEGFR SNPs. Results from angiogenesis genotyping were then correlated with clinical outcome parameters. Globally, a total of 210 patients were enrolled. At multivariate analysis rs12434438 of HIF1α, rs2010963 of VEGF-A, and rs4604006 of VEGF-C were confirmed as independent predictive factors. At the combined analysis of significant SNPs, the presence of two favourable alleles of rs2010963 and rs4604006 of VEGF compared to only one or to none favourable alleles, was able to identify three separate patients populations with different time-to-progression (TTP) (10.8 vs. 5.6 vs. 3.7months, respectively; p < 0.0001) and overall survival (OS) (19.0 vs. 13.5 vs. 7.5months, respectively; p < 0.0001). Furthermore, the presence of the GG genotype of rs12434438 (HIF-1α) seemed able to select a population with a particularly poor outcome, independently from the clinical effect of the two VEGF SNPs (TTP: 2.6months, HR: 0.54, p = 0.0374; OS: 6.6months, p = 0.0061, HR: 0.43). Our findings show that polymorphism analysis of HIF-1α, VEGF, and VEGFR genes may represent a prognostic panel to better identify HCC patients who are more likely to benefit from sorafenib treatment.

Gene polymorphisms in VEGFA and COL2A1 are associated with response to inhaled corticosteroids in children with asthma.

Aim: The purpose of this study was to investigate the involvement of single-nucleotide polymorphisms in VEGFA, TBX21 and COL2A1 in the response to inhaled corticosteroids in asthmatic children. Subjects & methods: Children with mild-to-moderate asthma were enrolled in the study. The SEQUENOM MassARRAY method was used to sequence 27 SNP genotypes. By ranking the data from smallest to largest, we could infer whether the change in distribution of forced expiratory volume in one second/forced vital capcacity (FEV1/FVC) and fractional exhaled nitric oxide differed between genotype groups. Results:VEGFA rs3025039 T allele carriers had a smaller change in FEV1 than CC carriers (p=0.040), and in COL2A1 rs3809324, the frequency of T allele carriers was lower than that of GG carriers (p=0.048). rs3025039 was also associated with changes in FEV1/FVC (p=0.016). Conclusion:VEGFA and COL2A1 polymorphisms are significantly associated with the response to inhaled corticosteroids in asthmatic children.

Effect of gene polymorphisms in ADAM33, TGFβ1, VEGFA, and PLAUR on asthma in Thai population.

Most of the asthma susceptibility genes have demonstrated moderate effect. Gene-gene interaction may play a role in asthma. To investigate the genetic and gene-gene interaction effects of single nucleotide polymorphisms (SNPs) in the ADAM33, TGFβ1, VEGFA, and PLAUR genes on asthma in Thai population. Two hundred and fifty control and 250 asthmatic Thai subjects were recruited. Asthma was diagnosed based on symptoms and spirometry assessments using criteria outlined by the American Thoracic Society. Degrees of asthma severity were determined according to guidelines provided by the Global Initiative for Asthma. Asthmatic subjects were subcategorized into the low-severity (n = 106) and high-severity (n = 144) groups. Eleven SNPs in four genes were genotyped, including ADAM33 SNPs (rs528557/S2, rs598418, rs44707/ST+4), TGFβ1 SNPs (rs2241715, rs11466345), VEGFA SNPs (rs833069, rs3025010), and PLAUR SNPs (rs344781, rs344787, rs2239374, rs2239372). Association analyses between SNPs and asthma, and tests for gene-gene interaction were performed. The ADAM33 rs528557/S2 SNP was found to be associated with asthma according to the additive and dominant models. Comparison between the low-severity group and controls showed the VEGFA rs833069 SNP to be significantly associated with the low-severity group. No gene-gene interactions were observed in this study. The ADAM33 rs528557/S2 and the VEGFA rs833069 SNPs were associated with Thai asthmatics, as well as with other populations worldwide. Further studies are warranted to investigate the use these SNPs as biomarkers for establishing early diagnosis or for predicting future risk of asthma.

Impact of vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor (VEGFR) single nucleotide polymorphisms on outcome in gastroenteropancreatic neuroendocrine neoplasms.

Angiogenesis represents a key event in cancer development, leading to local invasion e metastatization, and might be considered a basic feature in gastroenteropancreatic neuroendocrine neoplasms (GEP-NENs) with a high expression of angiogenic molecules. We aimed to analyze the prognostic and predictive role of angiogenic factors in GEP-NENs through the analysis of single nucleotide polymorphisms (SNPs) of VEGF-A, VEGFR2 and VEGFR3. The genomic DNA of 58 consecutive patients with GEP-NENs treated at our Institution was extracted from peripheral blood. Two SNPs were identified respectively in VEGF-A (rs2010963G>C, rs699947A>C), VEGFR-2 (rs2305948C>T, rs1870377T>A), and VEGFR-3 (rs307821T>C, rs307826C>A) gene. Gene polymorphisms were determined by Real-Time PCR using TaqMan assays. Median age was 57 years (range 24–79 years); 32 patients were male and 77.5% of NENs were localized in the pancreas. The allele frequency of VEGFR-2 rs2305948T and of VEGF-A rs2010963C showed a trend of higher frequency than in general population (12.1% vs. 8.0% and 34.5% vs. 31.2%, respectively). Three out SNPs (VEGF-A rs699947C, VEGF-A rs2010963GC and VEGFR-3 rs307821C) showed a correlation with an increased risk of disease relapse. Moreover median PFS changes according to the presence of 0–1 SNPs (20.7% of cases; 61.9 months), 2 SNPs (25.9%; 49.2 months) and 3 SNPs (53.4%; 27.8 months) (p = 0.034). Results suggest, for the first time, that specific SNPs in VEGF-A and VEGFR-3 correlate with poor prognosis in GEP-NENs. The identification of this new prognostic factor might be helpful in order to optimize the management of these heterogeneous neoplasms.

Vascular Endothelial Growth Factor-A (VEGF-A) Single Nucleotide Polymorphisms and Endometriosis: Still a Controversial Issue:

Endometriosis is a chronic, multifactorial, polygenic gynecological disease. Endometrium has a high angiogenic potential and endometriotic lesions involve large areas with a rich blood supply. Angiogenesis is controlled by numerous inducers, including the vascular endothelial growth factor (VEGF) family. Endometrium undergoes cyclical growth and regression during the menstrual cycle, which depends on ovarian steroid levels and is a rich source of angiogenic growth factors, including VEGF-A. The VEGF-A gene is located on chromosome 6p21.3 and it is highly polymorphic with more than 20 different variants. This article critically reviews the published data concerning the relationships between some of the VEGF-A single nucleotide polymorphisms and the risk, pathogenesis and stage of endometriosis. Contrasting results have been published in the literature - probably due to the different ethnic background and the number of patients enrolled in clinical trials. However, the increasing interest in the use of anti...

Vascular Endothelial Growth Factor Gene Polymorphism Is Associated With Long-term Kidney Allograft Outcomes

IntroductionVascular endothelial growth factor (VEGF) regulates vasculogenesis in physiological and pathological states. We evaluated the role of VEGF single-nucleotide polymorphisms (SNPs) −1154 G/A, −2578 C/A, +936 C/T, and −2549 Ins/Del in chronic allograft nephropathy.MethodsBlood samples were collected before renal transplantation, and DNA was extracted. Genotyping of VEGF SNPs −1154 G/A (rs1570360), −2578 C/A (rs699947), +936 C/T (rs112005313), and −2549 Ins/Del (18bpindel) polymorphisms were carried out. Relative quantification of VEGF-A mRNA expression for 4 VEGF SNPs were quantified by the 2−ΔΔCt algorithm. Kidney allografts were categorized into graft loss (n = 98) and normally functioning (n = 174) groups. Genotype frequencies were calculated using additive, dominant, and recessive models. Hardy−Weinberg Equilibrium was assessed between outcome groups by standard procedure using χ2 analysis. The cumulative allograft survival was estimated by Kaplan−Meier analysis and compared among VEGF genotypes by the log-rank test. Study limitations were the lack of VEGF serum levels, donor-specific antigens, and protocol biopsies.ResultsThere was an association of AA (hazard ratio = 2.42, P = 0.0001) and CA (hazard ratio = 1.83, P = 0.009) genotypes of −2578 C/A SNP with graft loss. After adjustment for transplant-related covariates, associations of VEGF SNPs −2578 C/A and −2549 Ins/Del with graft failure were found to be significant. There was prolonged graft survival for cases with the CC genotype of VEGF −2578 C/A SNP. The carrier −2578*CC, −1154*GG, and +936*CC genotypes were shown to have a strongly protective association. There was no association with posttransplantation lymphomas.ConclusionRecipients of kidney allografts possessing low-producing VEGF genotypes are associated with less prolonged graft survival.

VEGF, VEGFR2 and GSTM1 polymorphisms in Outcome of Multiple Myeloma Patients in the Thalidomide Era

Introduction: Angiogenesis (AG) abnormalities are crucial in pathogenesis of multiple myeloma (MM), and give support to treat patients with antiangiogenic agents. However, patients with similar clinicopathological aspects may present distinct outcome under AG inhibitors treatment. Single nucleotide polymorphisms (SNPs) in genes involved in blood vessels formation may constitute a plausible explanation for this finding. The wild-type alleles of VEGF c.-2595C>A (rs699947), c.-1154G>A (rs1570360), c.-634G>C (rs2010963), c.*237C>T (rs3025039), VEGFR2 c.-906T>C (rs2071559) and c.889G>A SNPs (rs2305948) SNPs, and GSTM1 and GSTT1 genes determine higher production, transcriptional activity, binding efficiency or best-characterized regulator of VEGF. This study aimed to investigate the roles of VEGF c.-2595C>A, c.-1154G>A, c.-634G>C, c.*237C>T, VEGFR2 c.-906T>C, c.889G>A SNPs, and GSTM1 and GSTT1 genes in outcome of MM patients treated with thalidomide-based regimens. Patients and methods: The study comprised 102 MM patients diagnosed at the Haematology and Haemotherapy Centre of University of Campinas between June 2005 and June 2013. The tumor was diagnosed by standard criteriaand staged by International Staging System. Therapeutic regimens consisted in thalidomide combined with steroids and chemotherapy, followed or not by autologous steam cell transplantation (ASCT). Response was evaluated at the end of treatment using the International Myeloma Working Group guidelines. The follow-up of patients was performed with hematological, biochemical, and immunological exams. The end of the study was February 2016. Genotypes of VEGF, VEGFR2 SNPs, and GSTM1 and GSTT1 genes were analyzed in genomic DNA by polymerase chain reaction based methods. The pairwise linkage disequilibrium (LD) was performed to ensure that markers were appropriate for inclusion in the VEGF and VEGFR2 haplotype estimates. The chi-square test and logistic regression model were used to identify variables influencing response to treatment. The Kaplan-Meier method, log-rank test and Cox hazards models served to assess the associations between event-free survival (EFS) and overall survival (OS). Results: Near half of patients enrolled in this study were male, and most of them were caucasians and with tumor at stages II or III. ASCT was performed after chemotherapy in near 40% of patients. LDs between VEGF and VEGFR2 SNPs were seen in study, and common haplotypes (frequency >1%) of the genes were included in further analyses. Patients with the wild-type allele of VEGF c.-2595C>A alone or plus the wild-type allele of VEGFR2 c.-906T>C SNPs, and the CGGC haplotype of all respective VEGF SNPs had 3.55, 9.91, and 3.86 more chances of achieving better response to therapy than others. The median follow-up time of 102 MM patients enrolled in the study was 43 months (range: 1-88). The estimated probabilities of 60-months EFS and OS were 24.5% and 48.1%, respectively. At 60 months of follow-up, patients with VEGFR2 c.889GG, VEGF c.-634GG plus VEGFR2 c.889GG, and VEGFR2 c.889GGplus GSTM1 present genotypes had 2.62, 2.64, and 2.80 more chances of presenting disease relapse or progression, and 2.21, 4.88, and 4.23 more chances of evolving to death in multivariate analysis, respectively. Conclusion: Our data present, for the first time, a preliminary evidence that VEGF c.-2595C>A, c.-1154G>A, c.-634G>C, c.*237C>T, VEGFR2 c.-906T>C, c.889G>A SNPs, and GSTM1 gene alter outcome of MM patients treated with thalidomide-based regimens. If these findings could be confirmed in multi-center studies with larger sample size, this might constitute a promise in assisting optimal patient choice for treatment with angiogenic agents. Financial support: São Paulo Research Foundation (FAPESP). DisclosuresNo relevant conflicts of interest to declare.

Germline Polymorphisms of the VEGF Pathway Predict Recurrence in Nonadvanced Differentiated Thyroid Cancer.

Tumor angiogenesis is determined by host genetic background rather than environment. Germline single nucleotide polymorphisms (SNPs) of the vascular endothelial growth factor (VEGF) pathway have demonstrated prognostic value in different tumors. Our main objective was to test the prognostic value of germline SNPs of the VEGF pathway in nonadvanced differentiated thyroid cancer (DTC). Secondarily, we sought to correlate analyzed SNPs with microvessel density (MVD). Multicenter, retrospective, observational study. Four referral centers. Blood samples were obtained from consecutive DTC patients. Genotyping was performed according to the TaqMan protocol, including 4 VEGF-A (-2578C>A, -460T>C, +405G>C, and +936C>T) and 2 VEGFR-2 (+1192 C>T and +1719 T>A) SNPs. MVD was estimated by means of CD34 staining. Rate of recurrent structural disease/disease-free survival (DFS). Difference in MVD between tumors from patients with different genotype. Two hundred four patients withstage I-II DTC (mean follow-up, 73 ± 64 months) and 240 patients with low- to intermediate-risk DTC (mean follow-up, 70 ± 60 months) were enrolled. Two "risk" genotypes were identified by combining VEGF-A SNPs -2578 C>A, -460 T>C, and +405 G>C. The ACG homozygous genotype was protective in both stage I-II (odds ratio [OR], 0.08; 95% confidence interval [CI], 0.01 to 1.43; P = 0.018) and low- to intermediate-risk (OR, 0.14; 95% CI, 0.01 to 1.13; P = 0.035) patients. The CTG homozygous genotype was significantly associated with recurrence in stage I-II (OR, 5.47; 95% CI, 1.15 to 26.04; P = 0.018) and was slightly deleterious in low- to intermediate-risk (OR, 3.39; 95% CI, 0.8 to 14.33; P = 0.079) patients. MVD of primary tumors from patients harboring a protective genotype was significantly lower (median MVD, 76.5 ± 12.7 and 86.7 ± 27.9, respectively; P = 0.024). Analysis of germline VEGF-A SNPs could empower a prognostic approach to DTC.

VEGF, VEGFR2 and GSTM1 Polymorphisms in Outcome of Multiple Myeloma Patients in the Thalidomide Era

Introduction : Angiogenesis (AG) abnormalities are crucial in pathogenesis of multiple myeloma (MM), and give support to treat patients with antiangiogenic agents. However, patients with similar clinicopathological aspects may present distinct outcome under AG inhibitors treatment. Single nucleotide polymorphisms (SNPs) in genes involved in blood vessels formation may constitute a plausible explanation for this finding. The wild-type alleles of VEGF c.-2595C>A (rs699947), c.-1154G>A (rs1570360), c.-634G>C (rs2010963), c.*237C>T (rs3025039), VEGFR2 c.-906T>C (rs2071559) and c.889G>A SNPs (rs2305948) SNPs, and GSTM1 and GSTT1 genes determine higher production, transcriptional activity, binding efficiency or best-characterized regulator of VEGF. This study aimed to investigate the roles of VEGF c.-2595C>A, c.-1154G>A, c.-634G>C, c.*237C>T, VEGFR2 c.-906T>C, c.889G>A SNPs, and GSTM1 and GSTT1 genes in outcome of MM patients treated with thalidomide-based regimens. Patients and methods : The study comprised 102 MM patients diagnosed at the Haematology and Haemotherapy Centre of University of Campinas between June 2005 and June 2013. The tumor was diagnosed by standard criteriaand staged by International Staging System. Therapeutic regimens consisted in thalidomide combined with steroids and chemotherapy, followed or not by autologous steam cell transplantation (ASCT). Response was evaluated at the end of treatment using the International Myeloma Working Group guidelines. The follow-up of patients was performed with hematological, biochemical, and immunological exams. The end of the study was February 2016. Genotypes of VEGF , VEGFR2 SNPs, and GSTM1 and GSTT1 genes were analyzed in genomic DNA by polymerase chain reaction based methods. The pairwise linkage disequilibrium (LD) was performed to ensure that markers were appropriate for inclusion in the VEGF and VEGFR2 haplotype estimates. The chi-square test and logistic regression model were used to identify variables influencing response to treatment. The Kaplan-Meier method, log-rank test and Cox hazards models served to assess the associations between event-free survival (EFS) and overall survival (OS). Results : Near half of patients enrolled in this study were male, and most of them were caucasians and with tumor at stages II or III. ASCT was performed after chemotherapy in near 40% of patients. LDs between VEGF and VEGFR2 SNPs were seen in study, and common haplotypes (frequency >1%) of the genes were included in further analyses. Patients with the wild-type allele of VEGF c.-2595C>A alone or plus the wild-type allele of VEGFR2 c.-906T>C SNPs, and the CGGC haplotype of all respective VEGF SNPs had 3.55, 9.91, and 3.86 more chances of achieving better response to therapy than others. The median follow-up time of 102 MM patients enrolled in the study was 43 months (range: 1-88). The estimated probabilities of 60-months EFS and OS were 24.5% and 48.1%, respectively. At 60 months of follow-up, patients with VEGFR2 c.889GG, VEGF c.-634GG plus VEGFR2 c.889GG, and VEGFR2 c.889GGplus GSTM1 present genotypes had 2.62, 2.64, and 2.80 more chances of presenting disease relapse or progression, and 2.21, 4.88, and 4.23 more chances of evolving to death in multivariate analysis, respectively. Conclusion : Our data present, for the first time, a preliminary evidence that VEGF c.-2595C>A, c.-1154G>A, c.-634G>C, c.*237C>T, VEGFR2 c.-906T>C, c.889G>A SNPs, and GSTM1 gene alter outcome of MM patients treated with thalidomide-based regimens. If these findings could be confirmed in multi-center studies with larger sample size, this might constitute a promise in assisting optimal patient choice for treatment with angiogenic agents. Financial support : Sao Paulo Research Foundation (FAPESP). Disclosures No relevant conflicts of interest to declare.

B19 - Impact of VEGF and VEGFR single nucleotide polymorphisms on outcome in gastroenteropancreatic neuroendocrine tumors

B19 - Impact of VEGF and VEGFR single nucleotide polymorphisms on outcome in gastroenteropancreatic neuroendocrine tumors

B27 - Pathogenic and prognostic role of VEGF and VEGFR single nucleotide polymorphisms in gastroenteropancreatic neuroendocrine neoplasms

B27 - Pathogenic and prognostic role of VEGF and VEGFR single nucleotide polymorphisms in gastroenteropancreatic neuroendocrine neoplasms