Additional file 1: Table S1. of Angiogenesis related genes NOS3, CD14, MMP3 and IL4R are associated to VEGF gene expression and circulating levels in healthy adults

VEGF plasma and expression levels per genotype. Table S2. VEGF plasma and expression levels per interaction condition. (DOCX 72 kb

Functional Epistatic Interaction between rs6046G>A in <em>F7</em> and rs5355C>T in <em>SELE</em> Modifies Systolic Blood Pressure Levels

<div><h3>Background</h3><p>Although numerous genetic studies have been performed, only 0.9% of blood pressure phenotypic variance has been elucidated. This phenomenon could be partially due to epistatic interactions. Our aim was to identify epistatic interaction(s) associated with blood pressure levels in a pre-planned two-phase approach.</p> <h3>Methods and Results</h3><p>In a discovery cohort composed of 3,600 French individuals, we found rs6046A allele in <em>F7</em> associated with decreased blood pressure levels (P≤3.7×10⁻³) and rs5355T allele in <em>SELE</em> associated with decreased diastolic blood pressure levels (P = 5×10⁻³). Both variants interacted in order to influence blood pressure levels (P≤0.048). This interaction was replicated with systolic blood pressure in 4,620 additional European individuals (P = 0.03). Similarly, in this replication cohort, rs6046A was associated with decreased blood pressure levels (P≤8.5×10⁻⁴). Furthermore, in peripheral blood mononuclear cells of a subsample of 90 supposed healthy individuals, we found rs6046A positively associated with <em>NAMPT</em> mRNA levels (P≤9.1×10⁻⁵), suggesting an eventual involvement of <em>NAMPT</em> expression in blood pressure regulation. Confirming this hypothesis, further transcriptomic analyses showed that increased <em>NAMPT</em> mRNA levels were positively correlated with <em>ICAM1</em>, <em>SELL</em>, <em>FPR1</em>, <em>DEFA1-3</em>, and <em>LL-37</em> genes expression (P≤5×10⁻³). The last two mRNA levels were positively associated with systolic blood pressure levels (P≤0.01) and explained 4% of its phenotypic variation.</p> <h3>Conclusion</h3><p>These findings reveal the importance of epistatic interactions in blood pressure genetics and give new insights for the role of inflammation in its complex regulation.</p> </div

Genetic variants associated with blood pressure.

*<p>: Log10 transformed values.</p><p>Beta coefficients are shown for significant associations.</p><p>Chr: chromosome, SNP: single nucleotide polymorphism, MAF: minor allele frequency, Beta: coefficient in the linear regression model, BP: blood pressure, P_meta: P meta-analysis, SBP: systolic blood pressure, DBP: diastolic blood pressure.</p

Blood pressure variations according to rs5355T allele in <i>SELE</i> and rs6046G/A genotypes in <i>F7</i> when compared to rs5355C allele in <i>SELE</i>.

<p>Only significant blood pressure variations are shown.</p><p>BP variations in individuals carrying rs5355T allele in <i>SELE</i> and rs6046GG in <i>F7</i> were compared with carriers of rs5355C allele in <i>SELE</i> and rs6046GG genotype in <i>F7</i>. BP variations in individuals carrying rs5355T allele in <i>SELE</i> and rs6046GA genotype in <i>F7</i> were compared with carriers of rs5355C allele in <i>SELE</i>, rs6046GA genotype in <i>F7</i>. BP variations in carriers of rs5355T allele in <i>SELE</i> and rs6046AA genotype in <i>F7</i> were compared with those carrying rs5355C allele in <i>SELE</i> and rs6046AA genotype in <i>F7</i>.</p><p>DBP: diastolic blood pressure, P*: p value for epistatic interaction model, SBP: systolic blood pressure, BP: blood pressure.</p

Summary of the study and hypothesis for rs5355C>T in <i>SELE</i> and rs6046G>A in <i>F7</i> interaction.

<p>rs6046A allele in <i>F7</i> was associated with decreased BP levels. rs5355C>T in <i>SELE</i> and rs6046G>A in <i>F7</i> interacted in order to alter SBP levels, rs6046A inverted the BP-lowering effect of rs5355T. rs6046A allele in <i>F7</i> was positively associated with increased <i>NAMPT</i> gene expression. <i>NAMPT</i> levels were positively correlated with <i>ICAM1</i>, <i>SELL</i>, <i>FPR1</i> and <i>DEFA1-3</i> genes expression. Only <i>DEFA1-3</i> and <i>LL-37</i> expressions were correlated and associated with SBP levels and explained 4% of its variation.</p

Pearson’s correlations between <i>NAMPT, ICAM1, SELL, FPR1, DEFA1-3</i> and <i>LL-37</i> genes expression.

<p>Only Significant correlations are shown (P≤5×10⁻³).</p><p>All genes expression were normalized to <i>POL2RA</i> mRNA levels.</p><p>r: Pearson’s correlation coefficient, P: P-value.</p

Characteristics of studied individuals.

<p>BMI: body mass index, BP: blood pressure, SBP: systolic blood pressure, DBP: diastolic blood pressure, MAF: minor allele frequency.</p

DataSheet_1_Biomarkers to assess the risk of bladder cancer in patients presenting with haematuria are gender-specific.docx

IntroductionHaematuria is a common red flag symptom of urinary tract cancer. Bladder cancer (BC) is the most common cancer to present with haematuria. Women presenting with haematuria are often underdiagnosed. Currently, no gender-specific tests are utilized in clinical practice. Considerable healthcare resources are needed to investigate causes of haematuria and this study was set up to help identify markers of BC. The aim of the study was to define biomarker algorithms in haematuria patients using an expanded panel of biomarkers to diagnose BC and investigate if the algorithms are gender-specific.Materials and MethodsA total of n=675 patients with a history of haematuria were recruited from Northern Ireland hospitals. Patients were collected on a 2:1 ratio, non-BC (control) n=474: BC n=201. A detailed clinical history, urine and blood samples were collected. Biomarkers, known to be involved in the pathobiology underlying bladder carcinogenesis were investigated. Biomarkers differentially expressed between groups were investigated using Wilcoxon rank sum and linear regression.ResultsBiomarkers were gender specific. Two biomarker-algorithms were identified to triage haematuria patients; male – u_NSE, s_PAI-1/tPA, u_midkine, u_NGAL, u_MMP-9/TIMP-1 and s_prolactin (u=urine; s=serum); sensitivity 71.8%, specificity 72.8%; AUROC 0.795; and female urine biomarkers - IL-12p70, IL-13, midkine and clusterin; sensitivity 83.7%, specificity 79.7%; AUROC 0.865. Addition of the clinical variable infection to both algorithms increased both AUROC to 0.822 (DeLong p=0.014) and to 0.923 (DeLong p=0.004) for males and females, respectively. Combining clinical risk factors with biomarker algorithms would enable application of the algorithms to triage haematuria patients.ConclusionUsing gender-specific biomarker algorithms in combination with clinical risks that are associated with BC would allow clinicians to better manage haematuria patients and potentially reduce underdiagnosis in females. In this study, we demonstrate, for the first time, that blood and urine biomarkers are gender-specific when assessing risk of BC in patients who present with blood in their urine. Combining biomarker data with clinical factors could improve triage when referring patients for further investigations.</p

Presentation1_Design and selection of anti-PD-1 single-domain antibody and tumor necrosis factor superfamily ligands for an optimal vectorization in an oncolytic virus.PPTX

Arming oncolytic viruses with transgenes encoding immunomodulators improves their therapeutic efficacy by enhancing and/or sustaining the innate and adaptive anti-tumoral immune responses. We report here the isolation, selection, and vectorization of a blocking anti-human PDL1 single-domain antibody (sdAb) isolated from PDL1-immunized alpacas. Several formats of this sdAb were vectorized into the vaccinia virus (VV) and evaluated for their programmed cell death protein 1 (PD1)/PD1 ligand (PDL1) blocking activity in the culture medium of tumor cells infected in vitro. In those conditions, VV-encoded homodimeric sdAb generated superior PDL1 blocking activity compared to a benchmark virus encoding full-length avelumab. The sdAb was further used to design simple, secreted, and small tumor necrosis factor superfamily (TNFSF) fusions with the ability to engage their cognate receptors (TNFRSF) only in the presence of PDL1-positive cells. Finally, PDL1-independent alternatives of TNFRSF agonists were also constructed by fusing different variants of surfactant protein-D (SP-D) oligomerization domains with TNFSF ectodomains. An optimal SP-D–CD40L fusion with an SP-D collagen domain reduced by 80% was identified by screening with a transfection/infection method where poxvirus transfer plasmids and vaccinia virus were successively introduced into the same cell. However, once vectorized in VV, this construct had a much lower CD40 agonist activity compared to the SP-D–CD40L construct, which is completely devoid of the collagen domain that was finally selected. This latest result highlights the importance of working with recombinant viruses early in the payload selection process. Altogether, these results bring several complementary solutions to arm oncolytic vectors with powerful immunomodulators to improve their immune-based anti-tumoral activity.</p

Pro- and anti-angiogenic VEGF mRNAs in autoimmune thyroid diseases

<p>The aim of this study was to assess the relationships between five different splice isoforms of VEGF mRNA and its plasma levels in individuals treated for autoimmune thyroid diseases (AITD); mainly Graves’ disease (GD) and Hashimoto’s thyroiditis (HT). In a population from Tunisia, levels of thyroid hormones and antibodies were quantified simultaneously with plasma VEGF and VEGF mRNA isoforms after a period of 6 months of patients’ treatment. Plasma VEGF was measured in 110 AITD patients (21 GD and 89 HT patients). VEGF isoforms (VEGF121, VEGF165, VEGF145 and VEGF189 pro-angiogenic isoforms and VEGF165b anti-angiogenic isoform) in peripheral blood mononuclear cells were quantified in 71 patients (20 GD and 51 HT patients) and 86 healthy controls. Decreased levels of VEGF189 mRNA were observed in AITD compared to controls. VEGF165 was increased in GD patients compared to controls and the VEGF165b was increased in HT patients compared to GD. We observed increased levels of VEGF165b in hypothyroid AITD patients after treatment. We have also shown that the VEGF145 isoform levels were determined by FT4 in all patients and by the thyroid status after 6 months of treatment only in HT patients. An association was observed for VEGF165 mRNA levels with anti-TPO antibodies in all patients. Finally, FT4 was associated with VEGF plasma levels but only in healthy controls. In conclusion, this descriptive study highlights the specificity of VEGF mRNA isoforms in AITD, a fact underlining the need for novel clinical trials and the development of personalised theranostic approaches.</p