Kaplan-Meier curves for OS rates.

<p>OS rates were compared between CC and any G (GC or GG) for the SNP <i>IL-6</i>–174 in (A) all patients, (B) in not <i>MYCN</i> amplified patients, (C) in high-risk patients and (D) not <i>MYCN</i> amplified high-risk patients.</p

SNP genotype correlation to <i>IL-6</i> gene expression and NB outcome.

<p><i>In silico</i> and qRT-PCR analysis of <i>IL-6</i> mRNA expression in (A) 198 LCLs and (B) 31 NB tumors, respectively, stratified according to the SNP <i>IL-6</i>–174. (C) Luciferase report gene assay carried out in HEK293 cells. Data shown in percentage are the mean ± SD from three independent transfection experiments, each done in triplicate and compared with promoter less control. Kaplan-Meier analysis is shown, with individuals grouped by median of expression of <i>IL-6</i> for OS and Progression Free Survival (PFS) rates in (D and E) 88 NB patients and (F) only PFS for 102 INSS stage 4 patients with <i>MYCN</i> not amplified. OS data of Seeger dataset were not available.</p

Characteristics of NB patients stratified per <i>IL-6</i> -176 (G>C) SNP genotype.

<p>N.A.  =  not available.</p>a<p>p-values and ORs from comparison of allelic frequencies.</p>b<p>p-values and ORs from comparison of genotype frequencies (GG/GC vs CC).</p>c<p>p-values and ORs from comparison of stage 1+2 patients vs stage 3+4 patients.</p

Case-Control study of <i>IL-6</i>–176 (G>C) SNP.

a<p>P-value and OR obtained by Armitage's trend test.</p

Test for independent statistical significance of −174 <i>IL-6</i> SNP after adjustment for NB risk factors.

<p>HR, hazard ratio; CI, confidence interval; C-index, concordance index.</p>a<p>C-index calculated including in the model only the NB risk factor (Age or MYCN or INSS stage 4).</p>b<p>C-index calculated including in the model the NB risk factor and <i>IL-6</i> SNP.</p

Impact of Interleukin-6 –174 G>C Gene Promoter Polymorphism on Neuroblastoma

<div><p>Background</p><p>Common variants in DNA may predispose to onset and progression of neuroblastoma (NB). The genotype GG of single nucleotide polymorphism (SNP) rs1800795 (−174 G>C) in interleukin (IL)-6 promoter has been associated with lower survival of high-risk NB.</p><p>Result</p><p>To evaluate the impact of <i>IL-6</i> SNP rs1800795 on disease risk and phenotype, we analyzed 326 Italian NB patients and 511 controls. Moreover, we performed <i>in silico</i> and quantitative Real Time (qRT)-PCR analyses to evaluate the influence of the SNP on gene expression in 198 lymphoblastoid cell lines (LCLs) and in 31 NB tumors, respectively. Kaplan-Meier analysis was used to verify the association between <i>IL-6</i> gene expression and patient survival. We found that <i>IL-6</i> SNP is not involved in susceptibility to NB development. However, our results show that a low frequency of genotype CC is significantly associated with a low overall survival, advanced stage, and high-risk phenotype. The <i>in silico</i> (<i>p</i> = 2.61×10⁻⁵) and qRT-PCR (<i>p</i> = 0.03) analyses showed similar trend indicating that the CC genotype is correlated with increased level of <i>IL-6</i> expression. In report gene assay, we showed that the −174 C variant had a significantly increased transcriptional activity compared with G allele (<i>p</i> = 0.0006). Moreover, Kaplan-Meier analysis demonstrated that high levels of <i>IL-6</i> are associated with poor outcome in children with NB in two independent gene expression array datasets.</p><p>Conclusions</p><p>The biological effect of SNP <i>IL-6</i>–174 G>C in relation to promotion of cancer progression is consistent with the observed decreased survival time. The present study suggests that SNP <i>IL-6</i>–174 G>C may be a useful marker for NB prognosis.</p></div

Label-Free Optical Marker for Red-Blood-Cell Phenotyping of Inherited Anemias

The gold-standard methods for anemia diagnosis are complete blood counts and peripheral-smear observations. However, these do not allow for a complete differential diagnosis as that requires biochemical assays, which are label-dependent techniques. On the other hand, recent studies focus on label-free quantitative phase imaging (QPI) of blood samples to investigate blood diseases by using video-based morphological methods. However, when sick cells are very similar to healthy ones in terms of morphometric features, identification of a blood disease becomes challenging even with QPI. Here, we introduce a label-free optical marker (LOM) to detect red-blood-cell (RBC) phenotypes, demonstrating that a single set of all-optical parameters can clearly identify a signature directly related to an erythrocyte disease through modeling each RBC as a biological lens. We tested this novel biophotonic analysis by proving that several inherited anemias, such as iron-deficiency anemia, thalassemia, hereditary spherocytosis, and congenital dyserythropoietic anemia, can be identified and sorted, thus opening a novel route for blood diagnosis on a completely different concept based on LOMs

rs6441201 risk allele at 3q25 correlates with increased <i>MLF1</i> expression and <i>MLF1</i> silencing results in decreased cell growth in neuroblastoma cells.

<p><b>(a)</b><i>MLF1</i> mRNA expression is significantly higher in neuroblastoma cell lines harboring one or more copies of the rs6441201 risk allele (A) compared to neuroblastoma cell lines homozygous for the protective allele (GG). <b>(b)</b> Silencing of RSRC1 or MLF1 expression using pooled siRNAs resulted in 50–90% reduced mRNA levels by real-time quantitative PCR in neuroblastoma cell lines. <b>(c)</b> Confirmation by Western blot of knockdown at the protein level for RSRC1 and MLF1 after siRNA mediated silencing in neuroblastoma cell lines. <b>(d-g)</b> siRNA mediated silencing of MLF1 results in significant growth inhibition of neuroblastoma cells compared to non-targeting control siRNA; no effect was observed upon silencing of RSRC1. Cell growth measured by real-time cell sensing system (RT-ces).</p

Discovery of neuroblastoma susceptibility loci at chromosome 3q25 and 4p16.

<p>Regional association plots of genotyped and imputed SNPs novel susceptibility loci. Plots were generated using LocusZoom. Y-axes represent the significance of association (-log10 transformed P values) and the recombination rate. SNPs are color-coded based on pair-wise linkage disequilibrium (r²) with indicated SNPs <b>(a)</b> 3q25 locus: rs6441201 shown in purple (3.01 x 10⁻⁷; Odds Ratio: 1.21, 95% C.I.: 1.12–1.30). <b>(b)</b> 4p16 locus: rs3796727 shown in purple (p = 5.25 x 10⁻⁹; Odds Ratio: 1.26, 95% C.I.: 1.16–1.36).</p

Statistically significant and replicated SNP associations at 3q25 and 4p16.

<p>Statistically significant and replicated SNP associations at 3q25 and 4p16.</p