Transcriptomic Analysis from Normal Glucose Tolerance to T2D of Obese Individuals Using Bioinformatic Tools.

Understanding the role of white adipose tissue (WAT) in the occurrence and progression of metabolic syndrome is of considerable interest; among the metabolic syndromes are obesity and type 2 diabetes (T2D). Insulin resistance is a key factor in the development of T2D. When the target cells become resistant to insulin, the pancreas responds by producing more insulin to try to lower blood glucose. Over time, this can lead to a state of hyperinsulinemia (high levels of insulin in the blood), which can further exacerbate insulin resistance and contribute to the development of T2D. In order to understand the difference between healthy and unhealthy obese individuals, we have used published transcriptomic profiling to compare differences between the WAT obtained from obese diabetics and subjects who are obese with normal glucose tolerance and insulin resistance. The identification of aberrantly expressed messenger RNA (mRNA) and the resulting molecular interactions and signaling networks is essential for a better understanding of the progression from normal glucose-tolerant obese individuals to obese diabetics. Computational analyses using Ingenuity Pathway Analysis (IPA) identified multiple activated signaling networks in obesity progression from insulin-resistant and normal glucose-tolerant (IR-NGT) individuals to those with T2D. The pathways affected are: Tumor Necrosis Factor (TNF), Extracellular signal-Regulated protein Kinase 1/2 ERK1/2, Interleukin 1 A (IL1A), Protein kinase C (Pkcs), Convertase C5, Vascular endothelial growth factor (Vegf), REL-associated protein (RELA), Interleukin1/1 B (IL1/1B), Triggering receptor expressed on myeloid cells (TREM1) and Nuclear factor KB1 (NFKB1) networks, while functional annotation highlighted Liver X Receptor (LXR) activation, phagosome formation, tumor microenvironment pathway, LPS/IL-1 mediated inhibition of RXR function, TREM1 signaling and IL-6 signaling. Together, by conducting a thorough bioinformatics study of protein-coding RNAs, prospective targets could be exploited to clarify the molecular pathways underlying the development of obesity-related type 2 diabetes.

Heavy and Chronic Cannabis Addiction does not Impact Motor Function: BOLD-fMRI Study.

OBJECTIVE: The aim of this paper is to demonstrate the impact of heavy and chronic cannabis use on brain potential functional control, reorganization, and plasticity in the cortical area. METHODS: 23 cannabis users were convened in 3 user's groups. The first group included 11 volunteers with an average of 15 joins/day; the second group included 6 volunteers with an average of 1.5 joins/day; the third group included 6 volunteers with an average of 2.8 joins/week. Besides, a 6 healthy volunteers (control group). All healthy and cannabis users underwent identical brain BOLD-fMRI assessment of the motor function. Besides, neuropsychological and full biological assessments were achieved. RESULTS: BOLD-fMRI maps of motor areas were obtained, including quantitative evaluation of the activations in the motor area. Besides, statistical analysis of various groups was achieved. CONCLUSION: Chronic cannabis addiction of varying use strength by groups of heavy, moderate, low dose, and zero doses are shown to have systematically equivalent effects on the control of brain motor function. Indeed, the BOLD-fMRI shows a remarkable sensitivity to minimal brain plasticity and reorganization of the functional motor control of the studied cortical area, and such varionation was not shown. Specific elucidation of the cannabis effect mechanisms in this unique function should clarify further protective pharmacological effects. This might illuminate the use of neuronal resources to prepare processes for pharmacological use and pharmaceutical forms. This suggests exploring any potential cannabis pharmaceutical form in diseases involving motor impairments.

DNAJB3 attenuates metabolic stress and promotes glucose uptake by eliciting Glut4 translocation.

Failure of the heat shock response is a key event that leads to insulin resistance and type 2 diabetes. We recently showed that DNAJB3 co-chaperone is downregulated in obese and diabetic patients and that physical exercise restores its normal expression with a significant improvement of the clinical outcomes. In 3T3-L1 adipocytes, DNAJB3 has a role in improving the sensitivity to insulin and glucose uptake. In co-immunoprecipitation assays, DNAJB3 interacts with both JNK1 and IKKß kinases. However, the functional impact of such interaction on their activities has not been investigated. Here, we assessed the effect of DNAJB3 on the respective activity of JNK1 and IKKß in cell-based assays. Using JNK1- and IKKß-dependent luciferase reporters, we show a marked decrease in luciferase activity by DNAJB3 in response to PMA and TNF-α that was consistent with a decrease in the translocation of p65/NF-κB to the nucleus in response to LPS. Furthermore, TNF-α-mediated IL-6 promoter activation and endogenous mRNA expression are significantly abrogated by DNAJB3 both in 3T3-L1 and C2C12 cells. The ability of DNAJB3 to mitigate ER stress and oxidative stress was also investigated and our data show a significant improvement of both forms of stress. Finally, we examined the effect of overexpressing and knocking down the expression of DNAJB3 on glucose uptake in C2C12 as well as the molecular determinants. Accordingly, we provide evidence for a role of DNAJB3 in promoting both basal and insulin-stimulated glucose uptake. Our finding reveals also a novel role of DNAJB3 in eliciting Glut4 translocation to the plasma membrane. These results suggest a physiological role of DNAJB3 in mitigating metabolic stress and improving glucose homeostasis and could therefore represent a novel therapeutic target for type 2 diabetes.

Evaluation of cationic channel TRPV2 as a novel biomarker and therapeutic target in Leukemia-Implications concerning the resolution of pulmonary inflammation.

Patients treated during leukemia face the risk of complications including pulmonary dysfunction that may result from infiltration of leukemic blast cells (LBCs) into lung parenchyma and interstitium. In LBCs, we demonstrated that transient receptor potential vanilloid type 2 channel (TRPV2), reputed for its role in inflammatory processes, exhibited oncogenic activity associated with alteration of its molecular expression profile. TRPV2 was overexpressed in LBCs compared to normal human peripheral blood mononuclear cells (PBMCs). Additionally, functional full length isoform and nonfunctional short form pore-less variant of TRPV2 protein were up-regulated and down-regulated respectively in LBCs. However, the opposite was found in PBMCs. TRPV2 silencing or pharmacological targeting by Tranilast (TL) or SKF96365 (SKF) triggered caspace-mediated apoptosis and cell cycle arrest. TL and SKF inhibited chemotactic peptide fMLP-induced response linked to TRPV2 Ca2+ activity, and down-regulated expression of surface marker CD38 involved in leukemia and lung airway inflammation. Challenging lung airway epithelial cells (AECs) with LBCs decreased (by more than 50%) transepithelial resistance (TER) denoting barrier function alteration. Importantly, TL prevented such loss in TER. Therefore, TRPV2 merits further exploration as a pharmacodynamic biomarker for leukemia patients (with pulmonary inflammation) who might be suitable for a novel [adjuvant] therapeutic strategy based on TL.

TCF7L2 is reproducibly associated with type 2 diabetes in various ethnic groups: a global meta-analysis.

TCF7L2 variants have been consistently associated with type 2 diabetes (T2D) in populations of different ethnic descent. Among them, the rs7903146 T allele is probably the best proxy to evaluate the effect of this gene on T2D risk in additional ethnic groups. In the present study, we investigated the association between the TCF7L2 rs7903146 polymorphism and T2D in Moroccans (406 normoglycemic individuals and 504 T2D subjects) and in white Austrians (1,075 normoglycemic individuals and 486 T2D subjects). Then, we systematically reviewed the association of this single nucleotide polymorphism (SNP) with T2D risk in a meta-analysis, combining our data with data from previous studies. The allelic odds ratios (ORs) for T2D were 1.56 [1.29-1.89] (p = 2.9 x 10(-6)) and 1.52 [1.29-1.78] (p = 3.0 x 10(-7)) in Moroccans and Austrians, respectively. No heterogeneity was found between these two different populations by Woolf test (chi (2) = 0.04, df = 1, p = 0.84). We found 28 original published association studies dealing with the TCF7L2 rs7903146 polymorphism in T2D. A meta-analysis was then performed on 29,195 control subjects and 17,202 cases. No heterogeneity in genotypic distribution was found (Woolf test: chi (2) = 31.5, df = 26, p = 0.21; Higgins statistic: I2 = 14.1%). A Mantel-Haenszel procedure was then performed to provide a pooled odds ratio (OR) of 1.46 [1.42-1.51] (p = 5.4 x 10(-140)). No publication bias was detected, using the conservative Egger's regression asymmetry test (t = -1.6, df = 25, p = 0.11). Compared to any other gene variants previously confirmed by meta-analysis, TCF7L2 can be distinguished by its tremendous reproducibility of association with T2D and its OR twice as high. In the near future, large-scale genome-wide association studies will fully extend the genome coverage, potentially delivering other common diabetes-susceptibility genes like TCF7L2.

GAD2 on chromosome 10p12 is a candidate gene for human obesity.

The gene GAD2 encoding the glutamic acid decarboxylase enzyme (GAD65) is a positional candidate gene for obesity on Chromosome 10p11-12, a susceptibility locus for morbid obesity in four independent ethnic populations. GAD65 catalyzes the formation of gamma-aminobutyric acid (GABA), which interacts with neuropeptide Y in the paraventricular nucleus to contribute to stimulate food intake. A case-control study (575 morbidly obese and 646 control subjects) analyzing GAD2 variants identified both a protective haplotype, including the most frequent alleles of single nucleotide polymorphisms (SNPs) +61450 C>A and +83897 T>A (OR = 0.81, 95% CI [0.681-0.972], p = 0.0049) and an at-risk SNP (-243 A>G) for morbid obesity (OR = 1.3, 95% CI [1.053-1.585], p = 0.014). Furthermore, familial-based analyses confirmed the association with the obesity of SNP +61450 C>A and +83897 T>A haplotype (chi(2) = 7.637, p = 0.02). In the murine insulinoma cell line betaTC3, the G at-risk allele of SNP -243 A>G increased six times GAD2 promoter activity (p < 0.0001) and induced a 6-fold higher affinity for nuclear extracts. The -243 A>G SNP was associated with higher hunger scores (p = 0.007) and disinhibition scores (p = 0.028), as assessed by the Stunkard Three-Factor Eating Questionnaire. As GAD2 is highly expressed in pancreatic beta cells, we analyzed GAD65 antibody level as a marker of beta-cell activity and of insulin secretion. In the control group, -243 A>G, +61450 C>A, and +83897 T>A SNPs were associated with lower GAD65 autoantibody levels (p values of 0.003, 0.047, and 0.006, respectively). SNP +83897 T>A was associated with lower fasting insulin and insulin secretion, as assessed by the HOMA-B% homeostasis model of beta-cell function (p = 0.009 and 0.01, respectively). These data support the hypothesis of the orexigenic effect of GABA in humans and of a contribution of genes involved in GABA metabolism in the modulation of food intake and in the development of morbid obesity.

Impaired histone deacetylases 5 and 6 expression mimics the effects of obesity and hypoxia on adipocyte function.

OBJECTIVE: The goal of the study was to investigate the role of histone deacetylases (HDACs) in adipocyte function associated with obesity and hypoxia. METHODS: Total proteins and RNA were prepared from human visceral adipose tissues (VAT) of human obese and normal weight subjects and from white adipose tissue (WAT) of C57Bl6-Rj mice fed a normal or high fat diet (HFD) for 16 weeks. HDAC activity was measured by colorimetric assay whereas the gene and protein expression were monitored by real-time PCR and by western blotting, respectively. RNA interference (RNAi) was used to silence the expression of genes in 3T3-L1 adipocytes. RESULTS: Total HDAC activity was decreased in VAT and WAT from obese individuals and from mice fed a HFD, respectively. The HDAC activity reduction was associated with decreased HDAC5/Hdac5 and HDAC6/Hdac6 expression in human and mice adipocyte fraction. Similarly, hypoxia hampered total Hdac activity and reduced the expression of Hdac5 and Hdac6 in 3T3-L1 adipocytes. The decrease of both Hdac5 and Hdac6 by hypoxia was associated with altered expression of adipokines and of the inducible cAMP early repressor (Icer), a key repressor that is defective in human and mice obesity. Silencing of Icer in adipocytes reproduced the changes in adipokine levels under hypoxia and obesity, suggesting a causative effect. Finally, modeling the defect of the two Hdacs in adipocytes by RNAi or selective inhibitors mimicked the effects of hypoxia on the expression of Icer, leading to impairment of insulin-induced glucose uptake. CONCLUSION: Hdac5 and Hdac6 expression are required for the adequate expression of Icer and adipocyte function. Altered adipose expression of the two Hdacs in obesity by hypoxia may contribute to the development of metabolic abnormalities.

Hypertension and type 2 diabetes: a cross-sectional study in Morocco (EPIDIAM Study).

BACKGROUND: In Morocco, there are no studies that focused on the hypertension and its associated risk factors through patients with type 2 diabetes. Different findings show that the frequency of type 2 diabetes has risen rapidly in Morocco. The main objective of this study was to assess the prevalence of hypertension and its associated risk factors among a group of patients with type 2 diabetes and to examine the level of control of hypertension among type 2 diabetic patients with hypertension. METHODS: A cross-sectional study was carried out on 525 type 2 diabetics in three Moroccan regions. The structured questionnaire was used to gather information on sociodemographic variables, history of hypertension, use of anti-hypertensive medications and duration of diabetes. Anthropometric measurements including weight and height were measured by trained staff. Blood pressure was measured using standardized sphygmomanometers. RESULTS: The prevalence of hypertension was 70.4%. The logistic regression indicated that hypertension was positively associated with age (p<10-4), BMI (p<0.0002) and duration of diabetes (p) CONCLUSION: Hypertension is a common co-morbidity among Moroccan diabetic patients with high rate of ignorance of hypertension among study subjects. The focus must be on patients and family education, counseling and behavioral interventions designed to modify lifestyle such as increasing physical activity and adopting recommended dietary changes, as well as compliance with medications.

Disease-specific health-related quality of life instruments among adults diabetic: A systematic review.

This paper provides a systematic review on health-related quality of life (HRQoL) measures in diabetic patients. For each included study, a description of the measure and its psychometric findings is provided. To evaluate these measures, a databases search (Medline, Scopus and Proqolid) was undertaken to identify relevant publications. Instruments were assessed according to predefined inclusion and exclusion criteria. Sixteen instruments met the inclusion criteria among 1049 references produced: appraisal of diabetes scale (ADS), audit of diabetes-dependent quality of life (ADDQoL), diabetes-39 (D-39), diabetes care profile (DCP), diabetes distress scale (DDS), diabetes health profile (DHP-1, DHP-18), diabetes impact measurement scales (DIMS), diabetes quality of life measure (DQOL), diabetes quality of life clinical trial questionnaire-revised (DQLCTQ-R), diabetes-specific quality of life scale (DSQOLS), elderly diabetes burden scale (EDBS), insulin delivery system rating questionnaire (IDSRQ), quality of life with diabetes questionnaire (LQD), problem areas in diabetes scale (PAID), questionnaire on stress in diabetic patients-revised (QSD-R) and well-being enquiry for diabetics (WED). All those instruments have been developed in northern countries. The shortest instrument (ADS) has seven items and the longest (IDSRQ) has 67 items. ADDQoL was widely translated followed by DHP and PAID. Only authors of ADS and DIMS have not involved patients in the construction of instruments. The authors of instruments: ADS, ADDQoL, DHP, D-39, and PAID reported the item-total correlation which is ranged from 0.28 to 0.84. The ADS, DQOL, EDBS, IDSRQ, LQD, PAID, QSD-R, and WED have been assessed for test-retest reliability which varies between 0.27 and 0.99. The DQLCTQ-R, DQOL and IDSRQ were not subjected to factor analysis. Responsiveness was assessed in PAID with effect sizes and ranged from 0.32 to 0.65 for interventions. Four domains were responsive to clinical change in metabolic control in DQLCTQ-R. The other instruments were not been formally assessed for responsiveness. This review found evidence that the instruments: ADDQoL, D-39, DDS, DHP1/18, DSQOLS, EDBS and QSD-R had adequate psychometric properties. For future research, responsiveness should be a priority and further study is also required to examine the effect of ethnicity and to determine the validity of these scales in developing countries.

Post genome-wide association studies of novel genes associated with type 2 diabetes show gene-gene interaction and high predictive value.

BACKGROUND: Recently, several Genome Wide Association (GWA) studies in populations of European descent have identified and validated novel single nucleotide polymorphisms (SNPs), highly associated with type 2 diabetes (T2D). Our aims were to validate these markers in other European and non-European populations, then to assess their combined effect in a large French study comparing T2D and normal glucose tolerant (NGT) individuals. METHODOLOGY/PRINCIPAL FINDINGS: In the same French population analyzed in our previous GWA study (3,295 T2D and 3,595 NGT), strong associations with T2D were found for CDKAL1 (OR(rs7756992) = 1.30[1.19-1.42], P = 2.3x10(-9)), CDKN2A/2B (OR(rs10811661) = 0.74[0.66-0.82], P = 3.5x10(-8)) and more modestly for IGFBP2 (OR(rs1470579) = 1.17[1.07-1.27], P = 0.0003) SNPs. These results were replicated in both Israeli Ashkenazi (577 T2D and 552 NGT) and Austrian (504 T2D and 753 NGT) populations (except for CDKAL1) but not in the Moroccan population (521 T2D and 423 NGT). In the overall group of French subjects (4,232 T2D and 4,595 NGT), IGFBP2 and CXCR4 synergistically interacted with (LOC38776, SLC30A8, HHEX) and (NGN3, CDKN2A/2B), respectively, encoding for proteins presumably regulating pancreatic endocrine cell development and function. The T2D risk increased strongly when risk alleles, including the previously discovered T2D-associated TCF7L2 rs7903146 SNP, were combined (8.68-fold for the 14% of French individuals carrying 18 to 30 risk alleles with an allelic OR of 1.24). With an area under the ROC curve of 0.86, only 15 novel loci were necessary to discriminate French individuals susceptible to develop T2D. CONCLUSIONS/SIGNIFICANCE: In addition to TCF7L2, SLC30A8 and HHEX, initially identified by the French GWA scan, CDKAL1, IGFBP2 and CDKN2A/2B strongly associate with T2D in French individuals, and mostly in populations of Central European descent but not in Moroccan subjects. Genes expressed in the pancreas interact together and their combined effect dramatically increases the risk for T2D, opening avenues for the development of genetic prediction tests.