Growth Factor Binding Protein Research Articles

Insulin like growth factor binding protein 2 (IGFBP-2) for risk prediction in patients with severe aortic stenosis undergoing Transcatheter Aortic Valve Implantation (TAVI)

IntroductionSevere aortic stenosis (AS) caused by degenerative calcification is the most frequent acquired valvular heart disease worldwide and mortality rates are considerably high. Transcatheter Aortic Valve Implantation (TAVI) is a well-established method for valve replacement in high risk patients with AS. However, there is a lack of reliable predictors for patients undergoing TAVI since commonly used scores were developed for surgical populations. Materials and methods208 patients subjected to TAVI were included in this study. Plasma samples were obtained before TAVI and were evaluated for IGFBP-2 using commercially available ELISA kits. IGFBP-2 levels were analyzed for their ability for risk prediction after TAVI. ResultsIGFBP-2 levels measured before TAVI correlated significantly with left ventricular ejection fraction, EUROSCORE and other functional and prognostic parameters like the 6-minute walking test. When patients were retrospectively divided in two groups with a cut-off of serum IGFBP-2 levels of 275 ng/ml, IGFBP-2 was a strong predictor for 30-day and one-year mortality (3% vs. 11%, p = 0.05 and 18.2% vs. 46.2%; p < 0.001 respectively). Compared to an EUROSCORE above 20 or an STS score cut-off above 8, IGFBP-2 plasma levels above 275 ng/ml outperformed the established risk score for prediction of one-year mortality as assessed by NRI (0.65 95% CI 0.37–0.94; p < 0.001 and 0.54 95% CI 0.25–0.82; p < 0.001, respectively). ConclusionsOur results indicate that IGFBP-2 could serve as new outcome predictor for patients undergoing TAVI procedure. By providing additional information to the commonly used EUROSCORE, IGFPB-2 analysis could further assist Heart Team decision making.

Association between the methylation of six apoptosis‑associated genes with autism spectrum disorder.

Excessive apoptosis hinders the process of brain maturation and is regarded as one of the principal risk factors for the development of autism spectrum disorder (ASD). The aim of the present study was to investigate the association between the methylation of six apoptosis‑associated genes [transforming growth factor β 1 (TGFB1), BCL2 associated X, apoptosis regulator, insulin like growth factor binding protein 3, protein kinase C β 1, presenilin 2 and C‑C motif chemokine ligand 2] and ASD. Using quantitative methylation‑specific polymerase chain reaction technology, DNA methylation levels were detected in 42 autistic and 26 control subjects. The logistic regression analysis results demonstrated that of the six genes, only TGFB1 was significantly hypomethylated in peripheral blood samples from children with autism compared with control samples (mean percentage of methylated reference, 0.011% vs. 0.019%; age‑adjusted P=0.028). In addition, TGFB1 methylation was identified to be positively associated with the interaction ability score from the Autism Behavior Checklist (r=0.452; P=0.035). These data suggested that decreased TGFB1 methylation may contribute to the development of ASD.

Brain-derived neurotrophic factor, insulin like growth factor-1 and inflammatory cytokine responses to continuous and intermittent exercise in patients with type 1 diabetes

AimsType 1 diabetes mellitus (T1DM) is an important risk factor for cognitive decline and motor dysfunction due to progressive muscular atrophy. Chronic hyperglycemia may be responsible for impaired vascular function, loss of muscle mass, and morphological abnormalities in the myocytes. For the proper functioning of the neuromuscular system, two crucial growth factors are necessary: brain-derived neurotrophic factor (BDNF) and insulin-like growth factor-1 (IGF-1), whose reduced expressions have been implicated in progressive neuropathy and muscle atrophy in patients with T1DM.The aim of the study was to compare the effects of two different exercise regimes (continuous and intermittent) on BDNF, IGF-1, blood glucose and inflammatory cytokine responses in young adults with and without Type 1 diabetes. MethodsFourteen patients (aged: 26.9 years) with T1DM and age-matched adults without diabetes participated in a 40 min continuous exercise (ExC, 50% of lactate threshold) and a high intensity intermittent exercise (ExInt, 120% of lactate threshold). During the study the patients performed self-monitoring of blood glucose levels (SMBG) under glycemic control. The effects of ExC and ExInt on BDNF, IGF-1, insulin like growth factor binding protein (IGFBP-3), insulin (INS), vascular endothelial growth factor (VEGF), transforming growth factor beta (TGF-β) and tumor necrosis factor alpha (TNF-α) were analyzed. ResultsBDNF and IGF-1 baseline serum levels were significantly lower in the T1DM patients compared to the healthy controls, but we found that ExInt and ExC significantly increase the secretion of BDNF and IGF-1 levels. Significant increases in BDNF and TGF-β levels, higher blood glucose decline, and a lower incidence of hypoglycaemia in ExInt compared to ExC were observed. Lower IGFBP-3 concentrations were revealed in T1DM patients in response to ExInt compared to ExC, suggesting a positive effect on IGF-1/IGFBP-3 ratio and the bioavailability of IGF-1. ConclusionsAccording to our results physical exercise has beneficial effects on serum BDNF and IGF-1 levels. A high-intensity intermittent exercise may be more effective at reducing the risk of exercise-induced glycaemic disorders in the T1DM patients as compared to continuous exercise.

Suboptimal Level of Bone-Forming Cells in Advanced Cirrhosis are Associated with Hepatic Osteodystrophy.

Bone loss is common in advanced cirrhosis, although the precise mechanisms underlying bone loss in cirrhosis are unknown. We studied the profile and functionality of bone‐forming cells and bone‐building proteins in bone marrow (BM) of individuals with cirrhosis (n = 61) and individuals without cirrhosis as normal controls (n = 50). We also performed dual energy X‐ray absorptiometry for clinical correlation. BM mesenchymal cells (MSCs) were analyzed for colony‐forming units‐fibroblasts and their osteogenic (fibronectin‐1 [FN1], insulin‐like growth factor binding protein 3 [IGFBP3], collagen type 1 alpha 1 chain [COL1A1], runt‐related transcription factor 2 [RUNX2], and alkaline phosphatase, liver [ALPL]) and adipogenic ( adiponectin, C1Q, and collagen domain containing [ADIPOQ], peroxisome proliferator‐activated receptor gamma [PPARγ], and fatty acid binding protein 4 [FABP4]) potentials. Colony‐forming units‐fibroblasts were lower in patients with cirrhosis (P = 0.002) than in controls. Cirrhotic BM‐MSCs showed >2‐fold decrease in osteogenic markers. Compared to controls, patients with cirrhosis showed fewer osteocytes (P = 0.05), osteoblasts, chondroblasts, osteocalcin‐positive (osteocalcin+) area, clusters of differentiation (CD)169+ macrophages (P < 0.001, each), and nestin+ MSCs (P = 0.001); this was more apparent in Child‐Turcotte‐Pugh (CTP) class C than A (P < 0.001). Multivariate logistic regression showed low nestin+ MSCs (P = 0.004) as a predictor of bone loss. Bone‐resolving osteoclasts were comparable among CTP groups, but >2‐fold decreased anti‐osteoclastic and increased pro‐osteoclastic factors were noted in patients with CTP C compared to CTP A. Bone‐building proteins (osteocalcin [P = 0.008], osteonectin [P < 0.001], and bone morphogenic protein 2 [P = 0.001]) were decreased while anti‐bone repair factors (fibroblast growth factor 23 [P = 0.015] and dipeptidyl peptidase 4 [P < 0.001]) were increased in BM and peripheral blood; this was more apparent in advanced cirrhosis. The dual energy X‐ray absorptiometry scan T score significantly correlated with the population of osteoblasts, osteocytes, MSCs, and CD169+ macrophages. Conclusion: Osteoprogenitor cells are substantially reduced in patients with cirrhosis and more so in advanced disease. Additionally, increased anti‐bone repair proteins enhance the ineffective bone repair and development of osteoporosis in cirrhosis. Hepatology Communications 2018;0:0‐0)

Quantitative screening of serum protein biomarkers by reverse phase protein arrays.

Screening biomarkers in serum samples for different diseases has always been of great interest because it presents an early, reliable, and, most importantly, noninvasive means of diagnosis and prognosis. Reverse phase protein arrays (RPPAs) are a high-throughput platform that can measure single or limited sets of proteins from thousands of patients' samples in parallel. They have been widely used for detection of signaling molecules involved in diseases, especially cancers, and related regulation pathways in cell lysates. However, this approach has been difficult to adapt to serum samples. Previously, we developed a sensitive method called the enhanced protein array to quantitatively measure serum protein levels from large numbers of patient samples. Here, we further refine the technology on several fronts: 1. simplifying the experimental procedure; 2. optimizing multiple parameters to make the assay more robust, including the support matrix, signal reporting method, background control, and antibody validation; and 3. establishing a method for more accurate quantification. Using this technology, we quantitatively measured the expression levels of 10 proteins: alpha-fetoprotein (AFP), beta 2 microglobulin (B2M), Carcinoma Antigen 15-3(CA15-3), Carcinoembryonic antigen (CEA), golgi protein 73 (GP73), Growth differentiation factor 15 (GDF15), Human Epididymis Protein 4 (HE4), Insulin Like Growth Factor Binding Protein 2 (IGFBP2), osteopontin (OPN) and Beta-type platelet-derived growth factor receptor (PDGFRB) from serum samples of 132 hepatocellular carcinoma (HCC) patients and 78 healthy volunteers. We found that 6 protein expression levels are significantly increased in HCC patients. Statistical and bioinformatical analysis has revealed decent accuracy rates of individual proteins, ranging from 0.617 (B2M) to 0.908 (AFP) as diagnostic biomarkers to distinguish HCC from healthy controls. The combination of these 6 proteins as a specific HCC signature yielded a higher accuracy of 0.923 using linear discriminant analysis (LDA), logistic regression (LR), random forest (RF) and support vector machine (SVM) predictive model analyses. Our work reveals promise for using reverse phase protein arrays for biomarker discovery and validation in serum samples.

Loss of high-mobility group N5 contributes to the promotion of human endometrial stromal cell decidualization.

PurposeHigh‐mobility group N (HMGN) proteins are the only non‐histone proteins that specifically bind within the nucleosome between core histones and DNA. Among them, HMGN5 is one of the candidates that could participate in mouse endometrial decidualization; however, the specific role of HMGN5 remains to be clarified in human endometrial stromal cells (HESCs).MethodsPrimary HESCs were isolated from hysterectomy specimens and incubated with or without 8‐bromo‐cyclic adenosine monophosphate (8‐br‐cAMP) and medroxyprogesterone acetate (MPA).ResultsWe demonstrated that HMGN5 expression in decidualized HESCs stimulated by 8‐br‐cAMP and MPA decreased significantly. The inhibition of HMGN5 expression by small interfering RNA (siRNA) induced the major decidual marker genes expression, including IGFBP1 (insulin‐like growth factor binding protein 1) and PRL (prolactin). In addition, microRNA‐542‐3p (miR‐542‐3p), which was identified as a regulatory miRNA of IGFBP1 during decidualization, was significantly suppressed by HMGN5 siRNA. However, the expression of HMGN5 was not alternated by miR‐542‐3p overexpression.ConclusionsThese findings suggest that the down‐regulation of HMGN5 plays a role in the promotion of human endometrial stromal decidualization and acts upstream of miR‐542‐3p.

Changes of the growth plate in children: 3-dimensional magnetic resonance imaging analysis.

Purpose This pilot study assessed changes in the growth plate and growth rates in children during a 6-month period.Methods The study included 31 healthy children (17 boys, 14 girls) under evaluation for growth retardation. Height, weight, bone age, insulin like growth factor-1 (IGF-1), and insulin like growth factor binding protein 3 (IGF-BP3) were measured at baseline and after 6 months. In addition, the diameter, thickness, and volume of the femoral and tibial growth plates were measured using magnetic resonance imaging.Results The mean bone age in boys and girls was 11.7 and 10.7 years, respectively. In boys, height (z score) (-0.2 vs. 0.0), weight (z score) (0.8 vs. 1.1), body mass index (BMI) (z score) (1.27 vs. 1.5), IGF-1 (ng/mL) (343.6 vs. 501.8), and IGF-BP3 (ng/mL) (5,088.5 vs. 5,620.0) were significantly higher after 6 months. In girls, height (z score) (-1.0 vs. -0.7), weight (z score) (-0.5 vs. 0.1), BMI (z score) (-0.02 vs. 0.3), IGF-1 (ng/mL) (329.3 vs. 524.6), and IGF-BP3 (ng/mL) (4,644.4 vs. 5,593.6) were also significantly higher after 6 months. In both sexes, the mean diameter and volume of the femoral and tibial growth plates were significantly increased 6 months later.Conclusion No significant correlation was found between changes in the growth plate and clinical parameters in children with growth retardation in this study, other than correlations of change in femoral diameter with weight and BMI. A larger, long-term study is needed to precisely evaluate the correlation between change in the growth plate and growth.

Protein Biomarkers of Cardiovascular Disease and Mortality in the Community.

BackgroundThe discovery of novel and highly predictive biomarkers of cardiovascular disease (CVD) has the potential to improve risk‐stratification methods and may be informative regarding biological pathways contributing to disease.Methods and ResultsWe used a discovery proteomic platform that targeted high‐value proteins for CVD to ascertain 85 circulating protein biomarkers in 3523 Framingham Heart Study participants (mean age, 62 years; 53% women). Using multivariable‐adjusted Cox models to account for clinical variables, we found 8 biomarkers associated with incident atherosclerotic CVD, 18 with incident heart failure, 38 with all‐cause mortality, and 35 with CVD death (false discovery rate, q<0.05 for all; P‐value ranges, 9.8×10−34 to 3.6×10−2). Notably, a number of regulators of metabolic and adipocyte homeostasis were associated with cardiovascular events, including insulin‐like growth factor 1 (IGF1), insulin‐like growth factor binding protein 1 (IGFBP1), insulin‐like growth factor binding protein 2 (IGFBP2), leptin, and adipsin. In a multimarker approach that accounted for clinical factors, growth differentiation factor 15 (GDF15) was associated with all outcomes. In addition, N‐terminal pro‐b‐type natriuretic peptide, C‐reactive protein, and leptin were associated with incident heart failure, and C‐type lectin domain family 3 member B (CLEC3B; tetranectin), N‐terminal pro‐b‐type natriuretic peptide, arabinogalactan protein 1 (AGP1), soluble receptor for advanced glycation end products (sRAGE), peripheral myelin protein 2 (PMP2), uncarboxylated matrix Gla protein (UCMGP), kallikrein B1 (KLKB1), IGFBP2, IGF1, leptin receptor, and cystatin‐C were associated with all‐cause mortality in a multimarker model.ConclusionsWe identified numerous protein biomarkers that predicted cardiovascular outcomes and all‐cause mortality, including biomarkers representing regulators of metabolic homeostasis and inflammatory pathways. Further studies are needed to validate our findings and define clinical utility, with the ultimate goal of improving strategies for CVD prevention.

Abstract 189: Examining how the extracellular environment alters gene expression in breast cancer

Abstract Breast cancer represents 25% of all cancers diagnosed in women globally with almost 1.7 million new cases annually. The average survival rate for patients with early stage breast cancer has increased, however the survival rate for those with metastatic breast cancer has not improved. Metastasis is the number one cause of death in breast cancer patients; it remains a scientific enigma, with no available early detection screen or targeted therapies available to patients. Therefore, understanding how and why cancer cells migrate is of significant biological and clinical interest. Accumulating evidence suggests the metastatic potential of breast cancer is influenced by the composition of the extracellular matrix (ECM). The ECM is composed of fibrous proteins and other non-cellular biomolecules and has a well-documented role in influencing tumour structure and behaviour. We are examining genes that code for several categories of proteins involved in regulating the composition of the ECM. These genes code for proteins that are involved in signal transduction from the cell membrane or for proteins that are secreted into the ECM. We are also examining genes that code for fibrous proteins, glycoproteins, transmembrane proteins, growth factors and proteases. To refine our panel, we used STRING [1], an online database, to examine the relationships between the proteins coded by our genes based on the experimental determination of protein co-expression and protein homology. Using qRT-PCR, we are examining the expression of our refined gene set in three different cell lines, (representing the major sub-categories of breast cancer) which have been cultured on collagen, fibronectin, laminin and stimulated by various growth factors. We extended our study to examine the expression pattern in cells that were maintained in 3-Dimensional culture over a period of 10 days in an attempt to mimic more closely the physiological process. We have determined that there is significant differential gene expression of integrins and growth factor binding proteins as we change the composition of the cells culture environment. This work is helping us to refine a signature set by identifying genes whose expression is dysregulated in response to changes in the stromal environment. We are moving now to determining whether dysregulation of the expression of our gene panel correlates with disease progression by examining matched normal and diseased patient tissue. This could provide the opportunity to generate a novel gene signature that may be used as a prognostic tool to predict invasive cancers. It may also provide information on the design of novel treatment therapies for patients with metastatic cancers. 1. Szklarczyk, D., et al., The STRING database in 2017: quality-controlled protein-protein association networks, made broadly accessible. Nucleic Acids Res, 2017. 45(D1): p. D362-d368. Citation Format: Joanne Nolan, Maeve Kiely, Colum P. Dunne, Aoife J. Lowery, Patrick A. Kiely. Examining how the extracellular environment alters gene expression in breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 189.

Decreased IL-1ra and NCAM-1/CD56 Serum Levels in Unmedicated Patients with Schizophrenia Before and After Antipsychotic Treatment.

ObjectiveSchizophrenia (SZ) has been associated with the inflammatory-related and immunological pathogenesis. This study investigates the aberration of cytokines in patients with SZ. MethodsThirty patients with SZ without antipsychotic treatment for at least two weeks participated. We measured the serum levels of fourteen cytokines at hospital admission and after 8-week antipsychotic treatment. Severity was measured by expanded version of 24-items brief psychiatric rating scale (BPRS-E). Repeated measure analyses of variance were conducted. ResultsThe interleukin-1 receptor antagonist (IL-1ra) was significantly decreased after 8-week antipsychotic treatment than those of before antipsychotic treatment (F=12.15, df=1/30, p=0.002). Neural cell adhesion molecule 1/CD56 (NCAM-1/CD56) was significantly decreased (F=6.61, df=1/30, p=0.016) among those with second-generation antipsychotics but not first-generation antipsychotics treatment. The changes of BPRS-E-manic and BPRS-E-anxiety scores correlated with the baseline IL-1ra (r=-0.393), IL-6 (r=-0.407), and insulin like growth factor binding protein 3 (r=-0.446). Additionally, the changes of BPRS-E and BPRS-E-negative scores correlated with the changes of brain-derived neurotrophic factor (r=0.372) and interferon-gamma (r=0.375). ConclusionOur study supports that IL-1ra and NCAM-1/CD56 may be considered as markers of developing SZ.

Cardiac and Vascular Surgery-Associated Acute Kidney Injury: The 20th International Consensus Conference of the ADQI (Acute Disease Quality Initiative) Group.

Acute kidney injury (AKI) occurs in 7% to 18% of hospitalized patients and complicates the course of 50% to 60% of those admitted to the intensive care unit, carrying both significant mortality and morbidity.[1][1] Even though many cases of AKI are reversible within days to weeks of occurrence, data

IGF-1 (Insulin like growth factor 1) and IGFBP2 (Insulin like growth factor binding protein 2) as potential biomarkers in differentiation between chronic pancreatitis (CP) and pancreatic adenocarcinoma (PDAC) in reference to pancreatic diabetes

IGF-1 (Insulin like growth factor 1) and IGFBP2 (Insulin like growth factor binding protein 2) as potential biomarkers in differentiation between chronic pancreatitis (CP) and pancreatic adenocarcinoma (PDAC) in reference to pancreatic diabetes

Lactobacillus plantarum Strain Ln4 Attenuates Diet-Induced Obesity, Insulin Resistance, and Changes in Hepatic mRNA Levels Associated with Glucose and Lipid Metabolism.

The prevalence of obesity and associated metabolic disorders, including diabetes and cardiovascular disease, is rapidly becoming a severe global health problem. Recent reports have suggested that the alteration of the gut ecosystem through the consumption of probiotics and fermented foods, such as yogurt and Kimchi, can significantly impact obesity and Type 2 diabetes (T2D)-related biomarkers. In this study, we screened over 400 strains of lactic acid bacteria (LAB) that were isolated from fermented foods to identify potent anti-obesogenic and diabetic probiotics in vitro. Of the strains tested, Lactobacillus plantarum Ln4 (Ln4), which was obtained from napa cabbage kimchi, significantly reduced lipid accumulation and stimulated glucose uptake in 3T3-L1 adipocytes. Oral administration of Ln4 reduced weight gain and epididymal fat mass in mice fed on a high-fat diet (HFD). Total plasma triglyceride level was significantly lower in mice that were treated Ln4 as compared with mice fed HFD. The protein levels of adipokines such as C-reactive protein (CRP), insulin-like growth factor binding proteins-3 (IGFBP-3), and monocyte chemoattractant protein-1 (MCP-1) decreased in white adipose tissues of Ln4-treated mice. Furthermore, these mice exhibited a significant reduction of insulin resistance index (HOMA-IR) and the improvement of glucose tolerance (OGTT) and insulin response (ITT) following Ln4 administration. This was associated with changes in several hepatic gene expressions (increased mRNA levels of IRS2, Akt2, AMPK, LPL, and reduced CD36) that regulate glucose and lipid metabolism. Taken together, these results indicate that in vitro and in vivo Ln4 treatment attenuates diet-induced obesity and T2D biomarkers, highlighting the potential of Ln4 as a therapeutic probiotic agent for metabolic disorders.

Effects of the Insulin‐like Growth Factor Axis and its Relationship in Nonsurgical Treatments in Patients with Lumbar Spinal Stenosis

BackgroundLumbar spinal stenosis (LSS) is commonly caused by degenerative arthritis of the spine and is characterized by narrowing of the spinal canal and/or neural foraminae. It is prevalent in older adults and leads to limitations in physical function and activities of daily living. Insulin‐like growth factor I (IGF‐I) and insulin‐like growth factor binding proteins (IGFBPs) have been associated with osteoporosis, chronic musculoskeletal diseases and metabolic disorders. A potential relationship between LSS and the IGF axis is currently unknown.PurposeThe purpose of this study was to examine the effects of three nonsurgical treatment programs in patients with LSS to determine potential relationships with the IGF axis and outcome measures of pain and physical function.MethodsThe study was a secondary analysis of stored plasma from a previously conducted randomized controlled trial. The final data set consisted of 102 participants (39 medical care (MC), 26 community‐based group exercise (GE), and 37 clinic‐based manual therapy and individual exercise (MT). The primary outcome measures were IGF‐I and IGFBP 1–7. Secondary outcome measures consisted of the Swiss Spinal Stenosis (SSS) Questionnaire, physical activity (defined as amount of time spent in light/moderate/vigorous activity (&gt; 1.5 METSs), and distance walked (meters) on a self‐paced walking test (SPWT). These outcome measures were collected at baseline, 2‐month, and 6‐month follow‐ups. IGF‐I and IGFBPs 1–7 were analyzed for within and between group differences across time using a two‐way repeated measures ANOVA. Correlation coefficients between the primary and secondary outcome measures were analyzed using Spearmans's rho. All data are means ± SD.ResultsThe two‐way ANOVA revealed a small group difference with IGFBP‐3 between GE and MT (6.81 ± 4.34 vs. 4.96 ± 1.79 ng/mL, respectively, p = 0.03), but not between GE and MC (6.81 ± 3.07 vs 5.45 ± 2.15 ng/mL, respectively, p = 0.18). No other differences between groups or other variables across time were observed (p &gt; 0.05). Using Spearman's rho we observed a small but significant positive correlation between baseline IGF‐I and IGFBP‐7 with baseline SSS (rs = 0.20, p = 0.04 and rs = 0.21, p = 0.03) but neither correlation was apparent at the 2‐ or 6‐month follow‐up. Lastly, a small positive correlation was also observed between baseline IGFBP‐1 and physical activity time at the 2‐month follow‐up (rs = 0.21, p = 0.04).ConclusionThe present study examined the systemic effects of three different nonsurgical treatment modalities on concentrations of IGF‐I and IGFBPs 1–7 in patients with LSS. Both physical activity and muscle stretch have been shown to favorably alter the IGF axis, potentially improving muscle function and reducing risk of degenerative diseases. However, the present study did not observe any effects suggesting improvement in the IGF axis from any of the three interventions.Support or Funding InformationFunding provided by a pilot grant from Wabtec Corporation, through the Center for Military Medicine ResearchThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Discovery and validation of a new biomarker for heart failure diagnostic

Diagnostic biomarkers for heart failure (HF) such as the natriuretic peptides (NPs) are widely used but have limitations. Innovative biomarkers could provide improved diagnostic performance. We launched a prospective case-control proteomic study and investigated for polypeptides specific to HF through a capillary electrophoresis-mass spectrometry (CE-MS) proteomic analysis. The putative biomarker was identified by Orbitrap liquid chromatography-MS, validated by western blot, then by ELISA using plasmas from multicentric international cohorts. A rat model of HF was tested for biomarker expression levels. We identified insulin like growth factor binding protein 2 (IGFBP2) as a new diagnostic biomarker for HF with a high sensitivity and specificity (AUC = 0.93; 95% CI, 0.89–0.96; P b 0.0001) in the local cohort and IGFBP2 levels provided an AUC of 0.943 (95% CI, 0.860–1.026) which gave a 87% sensitivity in AHF and 90% specificity at the cut off value previously determined in the discovery cohort, i.e. 556 ng/ml. ROC curve analysis of IGFBP2 and NTproBNP showed an AUC of 0.784 (95% CI, 0.744–0.820) for IGFBP2 and a significantly higher AUC of 0.927 (95% CI, 0.900–0.949) for NT-proBNP, P < 0.0001 in a Dutch cohort. In this cohort, the optimal cut off value for IGFBP2 gave a sensibility of 71% (95% CI, 66–76) and a specificity of 75% (95% CI, 65–83). IGFBP2 is a new biomarker to diagnose HF, which could be used to provide additional information to the NPs. Animals models, will help in the evaluation of the putative IGFBP2 regulated mechanisms in HF.

Abstract P327: Stress Overload and Methylation of Hypertension Related Genes

Background: Nearly 30% of U.S. adults are hypertensive with a high prevalence existing among Black Americans (42%). Exposure to psychosocial stress has been identified as a risk factor for hypertension and may explain part of the observed racial disparities. One pathway through which stress exposure may mechanistically alter biological functioning in a manner that affects risk of hypertension is via epigenetic modifications in hypertension-related genes, but this area remains understudied. Aims and Hypothesis: In the present study, we investigate the relationship between stress exposure and methylation of 25 genes that have been associated with blood pressure in Blacks in previous genetic research. We hypothesize that stress exposure is associated with DNA methylation of blood pressure related genes. Methods: We conducted an epigenetic association study using data from the ongoing InterGEN Study, a longitudinal investigation of the psychological, environmental, and genetic factors that contribute to hypertension risk in African American mothers (n=74) and their young children. Stress exposure was measured using the 24-item Stress Overload Scale (α=0.95), and a summary score was calculated by summing the Likert scale responses. Methylation was measured using the Illumina Infinium MethylationEPIC Beadchip, and M-values (i.e. log2 ratio of the intensities of methylated probe versus unmethylated probe) were calculated for association analysis. Linear regression was employed to assess the association between stress exposure and methylation in the a priori defined genes. Results: At p&lt;0.05, we identified 45 methylation sites associated with stress overload. These sites represented 17 of the 25 investigated genes. The most statistically significant sites were related to the genes: insulin like growth factor binding protein 3 ( IGFBP3 ), calcium voltage-gated channel subunit alpha1 H ( CACNA1H ), and solute carrier family 4 member 5 ( SLC4A5 ) (p=0.001, p=0.002, and p=0.006, respectively). Conclusion: This study provides biological insight into DNA methylation as a mechanism whereby exposure to psychosocial stress affects risk of hypertension in Black women. Additional replication studies with a larger sample size and includes men are needed to validate and generalize the findings. Future research should further investigate the biological mediators between DNA methylation and hypertension manifestation.

MiRNA‑27a regulates arthritis via PPARγ invivo and invitro.

The present study investigated the role of microRNA (miR)‑27a in the development of arthritis and its mechanism of action. Initially, collagen was used to develop an invivo rat model of arthritis. Changes in the miRs in the rats were analyzed. It was subsequently observed that miR‑27a expression was reduced in patients with arthritis, compared with the control group. In the present study an invitro miR‑27a overexpression model of arthritis was established and it was observed that miR‑27a increased the proliferation of osteoblast‑like cells invitro. miR‑27a overexpression promoted osteogenic differentiation, increased alkaline phosphatase (ALP) and osteoporosis (OST) content, induced insulin‑like growth factor binding protein-5 (IGFBP‑5) protein expression, reduced inflammation and suppressed peroxisome proliferator‑activated receptorγ (PPARγ) and matrix metalloproteinase-17 (MMP‑17) protein expression in arthritis. However, miR‑27a downregulation inhibited osteogenic differentiation, increased inflammation and PPARγ and MMP‑17 protein expression and suppressed ALP and OST content in an invitro model of arthritis. The PPARγ inhibitor reduced the function of miR‑27a downregulation on arthritis. Therefore the results of the present study revealed that miR‑27a regulates arthritis via PPARγ.

The clinical significance and expression of serum IGF-I and IGFBP-3 in the colorectal cancer

Objective To study the expression of insulin-like growth factor I(IGF-I )and insulin like growth factor binding protein -3(IGFBP-3)in the colorectal cancer and the relations between them. Methods A case-control study was conducted. We selected 113 patients with a pathologically conformed diagnostic colorectal cancer as the colorectal cancer (CRC) group, 37 patients with colorectal benign diseases as benign control (N-CRC) group, and other 76 healthy subjects as normal control (NC) group. CRC was divided into lymphatic metastasis (M-CRC) subgroup and no lymphatic metastasis (NM-CRC) subgroup and was also categorized by TNM staging. Concentrations of serum IGF-I and IGFBP-3 were measured by chemiluminescence method. SNK test and one-way ANOVA were used for comparison between the two groups and among multi-groups, respectively. Results A slightly decreasing trend of the concentrations of serum IGF-I was observed following the sequences of the NC, N-CRC and CRC groups. Moreover, the concentrations of serum IGF-I in T4 were significantly lower than that in(T1+ T2) and T3(F=6.279, P=0.003). The concentration of serum IGFBP-3 in the CRC group (3.13±1.57)μg/ml was significantly lower than that both in the N-CRC (3.42±1.32)μg/ml (F=3.04, P=0.09) and the Normal group (4.62± 1.10)μg/ml (F=10.88, P=0.001). The concentration of serum IGFBP-3 in the M-CRC was higher than that in NM-CRC(F=4.44, P<0.05). The ratio of IGF-I/IGFBP-3 in the CRC group (48.85±29.14) was remarkably higher than that both in the N-CRC group (42.38±12.58) (F=5.05, P=0.02) and the Normal group (42.46±16.24)(F=5.68, P=0.02). The value of diagnosis is general. The area under the ROC curve (AUC) of IGF-I was 0.63(95% CI 0.56-0.70), the sensitivity and specificity of IGF-I were 85.8% and 40.7% respectively when the critical value was 102.5 μg/ml.The area under the ROC curve (AUC) of IGFBP-3 was 0.72(95% CI 0.64-0.78), the sensitivity and specificity of IGFBP-3 were 78.8% and 62.8% respectively when the critical value was 3.32 μg/ml. Conclusions The concentrations of IGF-I and IGFBP-3 were low expressed in serum of patients with colorectal cancer. Meanwhile, they are correlated with the lymphatic metastasis and TNM staging.(Chin J Lab Med, 2017, 40: 871-875) Key words: Colorectal cancer; Insulin like growth factor I; Insulin like growth factor binding protein 3

Insulin‑like growth factor system in remission and flare of inflammatory bowel diseases.

INTRODUCTION Insulin‑like growth factor 1 (IGF‑1) is involved in the modulation of immunity and inflammation. It also plays a role in regulating the migration of endothelial cells and production of vasoactive agents. OBJECTIVES This study assessed the concentrations of IGF‑1 and insulin‑like growth factor-binding protein 3 (IGFBP‑3) and their relationships to disease activity in patients with inflammatory bowel disease (IBD). PATIENTS AND METHODS A total of 129 adult patients with IBD (69 with Crohn disease [CD] and 60 with ulcerative colitis [UC]) were involved in the study. The control group consisted of 31 healthy volunteers. Biochemical serum analyses were performed and the associations of IGF‑1 and IGFBP‑3 with inflammatory markers and disease activity were assessed. RESULTS IGF‑1 levels were decreased in patients with active UC compared with those with nonactive UC (mean [SD], 78.3 [22.7] ng/ml and 96.2 [24.5] ng/ml, respectively; P = 0.02) and controls (94.5 [26.5] ng/ml; P = 0.03). The IGF‑1 level was lower in patients with active CD compared with those with nonactive CD (mean [SD], 79.2 [24.9] ng/ml and 110.1 [43.4] ng/ml, respectively; P <0.001). The IGFBP‑3 level was lower in patients with active UC compared with those with nonactive UC (P = 0.04) and controls (P = 0.04). IGF‑1 correlated negatively with C‑reactive protein (CRP) levels (P <0.01), disease activity (P <0.05), and disease duration (P <0.05). IGFBP‑3 levels correlated negatively with CRP levels (P <0.05). CONCLUSIONS The IGF system is disrupted in patients with IBD. Systemic levels of the IGF axis components are related to disease activity and duration.

Estrogen-responsive gene networks in the teleost liver: What are the key molecular indicators?

An overarching goal of environmental genomics is to leverage sensitive suites of markers that are robust and reliable to assess biological responses in a range of species inhabiting variable environments. The objective of this study was to identify core groups of transcripts and molecular signaling pathways that respond to 17alpha-ethylinestadiol (EE2), a ubiquitous estrogenic contaminant, using transcriptome datasets generated from six independent laboratories. We sought to determine which biomarkers and gene networks were those most robust and reliably detected in multiple laboratories. Six laboratories conducted microarray analysis in pieces of the same liver from male fathead minnows exposed to ∼15ng/L EE2 for 96h. There were common transcriptional networks identified in every dataset. These included down-regulation of gene networks associated with blood clotting, complement activation, triglyceride storage, and xenobiotic metabolism. Noteworthy was that more than ∼85% of the gene networks were suppressed by EE2. Leveraging both these data and those mined from the Comparative Toxicogenomics Database (CTD), we narrowed in on an EE2-responsive transcriptional network. All transcripts in this network responded ∼±5-fold or more to EE2, increasing reliability of detection. This network included estrogen receptor alpha, transferrin, myeloid cell leukemia 1, insulin like growth factor 1, insulin like growth factor binding protein 2, and methionine adenosyltransferase 2A. This estrogen-responsive interactome has the advantage over single markers (e.g. vitellogenin) in that these entities are directly connected to each other based upon evidence of expression regulation and protein binding. Thus, it represents an interacting functional suite of estrogenic markers. Vitellogenin, the gold standard for estrogenic exposures, can show high individual variability in its response to estrogens, and the use of a multi-gene approach for estrogenic chemicals is expected to improve sensitivity. In our case, the coefficient of variation was significantly lowered by the gene network (∼67%) compared to Vtg alone, supporting the use of this transcriptional network as a sensitive alternative for detecting estrogenic effluents and chemicals. We propose that screening chemicals for estrogenicity using interacting genes within a defined expression network will improve sensitivity, accuracy, and reduce the number of animals required for endocrine disruption assessments.