bluuebunny/crossref_metadata_2025 huggingface dataset:
Maternal Exposure to Perfluorooctane Sulfonate (PFOS) Is Associated with Maternal Hyperglycaemia and Adverse Neonatal and Childhood Outcomes
Authors: MA RONALD C.W. , TAM CLAUDIA H.T. , YIU CHEUNG GRACE PUI , LOWE WILLIAM , METZGER BOYD E. , TAM WING H. , WONG CHRIS K.
Published: June 2018

Perfluorooctane sulfonate (PFOS) belongs to a class of endocrine-disrupting chemicals, perfluoroalkyl chemicals (PFCs), implicated in adiposity. Although supposedly phased out since 2002, its use remains widespread in Asia. We aim to examine the relationship between exposure to PFOS and maternal hyperglycaemia and metabolic outcomes in the offspring. We measured blood PFOS and other PFCs in archived samples taken at 24-32weeks gestation from mothers in the Hong Kong centre of the Hyperglycaemia and Adverse Pregnancy Outcome (HAPO) study between 2002-2004. All mothers underwent 75g OGTT and GDM was diagnosed according to the IADPSG/WHO 2013 criteria. Pregnancy outcomes, neonatal anthropometrics and childhood outcomes at 7 years were documented. PFCs were measured using high performance LC-MS-MS. We completed analysis of PFCs in 1,601 maternal samples, a subset of 99 cord blood samples, and 970 offspring at 7 years follow-up. There is strong correlation among PFOS and other PFCs in cord blood (ρ=0.51-0.74, p<0.001), as well as correlation with maternal levels (ρ=0.60, p<0.001). Ratio of cord blood to maternal PFOS was 0.60. Using regression analysis with adjustment for maternal age, BMI, and offspring gender, maternal PFOS showed association with maternal 1 hour glucose, 2 hour glucose, HbA1c, AUC during OGTT and GDM, though only the association with HbA1c remained significant using log-transformed PFOS (beta 0.0746 ± 0.0132, p=1.7x10-8). Log-transformed maternal PFOS was associated with higher birthweight, lower birth length, higher ponderal index and lower neonatal sum of skin fold thickness (beta -0.337 ± 0.085, p=7.1x 10-5), after adjustment for all covariates. Maternal PFOS during pregnancy was associated with lower height in offspring at age 7, but not offspring glycaemic parameters. Exposure to PFOS may be an important contributing factor to the epidemic of hyperglycaemia in pregnancy and childhood metabolic disorders in Asia.

Disclosure

R.C.W. Ma: Research Support; Self; AstraZeneca, Bayer AG, Merck Sharp & Dohme Corp., Pfizer Inc.. Advisory Panel; Self; Boehringer Ingelheim GmbH, Nippon Boehringer Ingelheim Co. Ltd.. C.H.T. Tam: None. G. Cheung: None. W. Lowe: None. B.E. Metzger: None. W.H. Tam: None. C.K. Wong: None.
Prognostic Cut Points for Prediabetes Glucose Levels and HbA1c Values in Predicting All-Cause Mortality—A Systematic Review and Meta-analysis
Authors: HUANG MINXUAN , GUJRAL UNJALI , HE SIRAN , JAGANNATHAN RAM , WEI JINGKAI , STAIMEZ LISA R. , VENKAT NARAYAN K.M.
Published: June 2018

Elevated but nondiabetic glucose levels, including impaired fasting glucose (IFG), 2-hour impaired glucose tolerance (IGT), and glycosylated hemoglobin A1c (HbA1c), are associated with increased risk for all-cause mortality. However, the glucose measures may differ in their abilities to predict adverse outcomes. The World Health Organization (WHO) and the American Diabetes Association (ADA) also differ in their definitions of IFG, and it remains unclear which definition best predicts future adverse outcomes. The objective of this study was to evaluate the prognostic value of different cut-points of prediabetic glucose measures for predicting all-cause mortality in individuals without diabetes. In databases including MEDLINE, PubMed, Embase, Clinicaltrials.gov, WHO International Clinical Trials Registry Platform, and Cochrane, we searched prospective cohort studies with minimum 3 years of follow-up, in adults without diabetes at enrollment. Data from eligible studies were pooled to synthesize results for each glucose measure. Random effect regression model was used to calculate pooled hazard ratio or relative risk for all-cause mortality. We screened over 4,000 abstracts and identified 170 eligible studies with mean follow-up of 10.2 years. Compared with individuals with normal glycemia, those with IFG defined by the WHO criteria had 1.14 times increased risk for all-cause mortality. There was no significant increase in mortality risk among those with IFG diagnosed by the ADA criteria. Individuals with IGT had 1.17 times higher risk for all-cause mortality compared with individuals with normal glycemia. HbA1c as low as 5.5% was associated with an increased risk for mortality. The WHO criteria for IFG seems to have better predictive value for all-cause mortality than the ADA criteria. Furthermore, individuals with HbA1c levels at 5.5% may have higher risk for mortality.

Disclosure

M. Huang: None. U. Gujral: None. S. He: None. R. Jagannathan: None. J. Wei: None. L.R. Staimez: None. K. Narayan: None.
The Maternal Impact of AdGHRKO Mice on Glucose Metabolism
Authors: WANG XIAOSHUANG , MI AI , LIU YANSHUANG , WANG DAN , RAN LIYUAN , WU YINGJIE
Published: June 2018

Genetically modified mouse models provide effective tools to explore the occurrence development and therapy of various diseases, especially the metabolic diseases such as diabetes. In line with the reported maternal inheritance of glucose homeostasis via maternally expressed gene activity, we found that a tissue-specific knockout exhibited maternal influence on insulin sensitivity, resulting in a lower liability for diabetes in the offsprings. Using the Cre/LoxP system, we constructed a mouse model of adipose tissue-specific growth hormone receptor (GHR) knockout (AdGHRKO). Unlike other tissue-specific GHR knockouts, AdGHRKO was found to be maternally inheritable. Compared with female AdGHRKO mice born from Flox mothers, female KO mice born from KO mothers displayed resistance to diet-induced obesity from a high-fat diet for 8 weeks. and the difference in body weight was exclusively due to a significant reduction in white fat tissue, especially from decreased subcutaneous deposition. Moreover, KO mice from KO mothers had better insulin sensitivity and glucose tolerance after high-fat treatment for 8 weeks. Our data suggests that the phenotype of metabolic stability in progeny is strongly affected by the maternal genotype. To support this, we also found that Flox mice born from KO mothers presented similar phenotypic traits (improved glucose tolerance and insulin resistance) of protection from high fat induced diabetes, which was comparable to the KO mice born from KO mothers. Our ongoing studies are to determine if this phenomenon is caused by alterations in maternal hormone levels or some unknown mitochondrial activities. Nonetheless, how adipose tissue specific ablation of GHR exerts its effects on metabolic syndromes through maternal cues is of high interest not only to pure genetic studies but also to medical practice.

Disclosure

X. Wang: None. A. Mi: None. Y. Liu: None. D. Wang: None. L. Ran: None. Y. Wu: None.
Characterization of a Type 2 Diabetes–Associated Islet-Specific Enhancer Cluster in STARD10 by Genome Editing of EndoC-ßH1 Cells
Authors: HU MING , GADUE PAUL , RUTTER GUY A.
Published: June 2018

Genome-wide association studies have identified >100 genetic loci associated with type 2 diabetes. We recently analysed one locus on chromosome 11q and mapped 5 causal variants in an islet-specific enhancer cluster within the STARD10 gene (Carrat, et al., Am J Hum Gen, 2017). Here, we aimed to understand the function of this cluster in beta cells and how causal variants influence enhancer activity. Using published ATAC-seq data (Varshney A, et al., PNAS, 2017) we analysed the transcriptional activity of 10 open chromatin regions (0.2-2 kb) using promoter-luciferase assay in EndoC-βH1 cells. Of these regions, HS1b displayed a 6.25-fold increase compared with empty vector [luciferase/renilla ratio: 6.25 ± 0.14; Student’s t-test, p<0.001; n=3]. Deletion of HS1b using CRISPR/Cas9 technology led to a significant reduction of glucose-stimulated insulin secretion (GSIS) (15/0.5 mM glucose, fold change: Control, 4.23 ± 0.41 vs. HS1b-del 2.21 ± 0.06; p<0.05; n=3) and ∼50% reduction of STARD10 gene expression (Fold: control 1 vs. HS1b-del, 0.486 ± 0.04; p<0.001, n=3). Chromosome conformation capture (4C and 3C) identified two enhancer regions, termed HS1 and HS5, as interacting with the causal variants including rs140130268. Both regions contain multiple defined CTCF binding sites. To test whether these CTCF binding sites create a chromatin loop defining an enhancer complex, we mutated four of the CTCF binding sites one by one. This resulted in a significant (p<0.05; n=3) impairment in GSIS assays [Control, 4.± 0.46, vs. HS5-1-Mu, 2.75 ± 0.07; HS5-2-Mu, 2.66 ± 0.13; HS1-1-Mu, 3.49 ± 0.02 and HS1-2-Mu, 3.47 ± 0.57]. Taken together, we show that the modification of enhancer elements at the STARD10 locus affects beta cell function. The causal variants, which are physically associated with both HS5 and HS1, are likely to exert their effects through the formation and activity of an enhancer-cluster complex which affects the expression of STARD10 and possibly other genes.

Disclosure

M. Hu: None. P. Gadue: None. G.A. Rutter: Research Support; Self; Servier.
Ancestral Differences in Hemoglobin A1C (A1C) Associations of Erythrocyte Genes Using NHLBI Trans-Omics for Precision Medicine (TOPMed) Whole Genome Sequence (WGS) Data
Authors: SARNOWSKI CHLOÉ , LEONG AARON , RAFFIELD LAURA ,   TOPMED DIABETES WORKING GROUP
Published: June 2018

The latest transethnic genome wide association study on A1C identified 22 loci that modify A1C independently of glycemia. These loci overlap genes implicated in erythrocyte phenotypes that vary in prevalence across populations. We used TOPMed WGS data to estimate ancestral differences in associations with A1C at these loci and 23 additional erythrocyte genes not previously known to be related to A1C.

We conducted WGS association analyses of A1C in 5224 nondiabetic individuals [2662 European ancestry (EA): Framingham Heart Study and Amish; 2562 African ancestry (AA): Jackson Heart Study] using age and sex-adjusted linear mixed-effect regression, and meta-analyzed cohort-specific results. We used Cochran heterogeneity and Fisher’s exact tests to assess ancestral differences in effect and Minor Allele Frequency (MAF).

We detected single variant associations in 11 genes/loci (P < 0.001; α = 0.05/45; ≥ 1 cohort; Table). Variants had ancestral differences in MAF (P < 1%) and ancestry-specific signals (AA: G6PD, HBB, HBA1; EA: ANK1, PIEZO1, SPTA1, SPTB, HBA2).

Genetic variation at erythrocyte genes exhibit ancestral differences in A1C effect and MAF. These variants may differ in their contributions to inter-individual A1C variation, which may impact glucose estimation and diabetes diagnosis by A1C, particularly in minority populations.

Disclosure

C. Sarnowski: None. A. Leong: None. L. Raffield: None.
Genome-Wide Association Study of Inflammatory Cytokines in Southwestern Native Americans Identifies Novel Association between WWOX and Serum TNF-? Levels
Authors: WEDEKIND LAUREN E. , WALTER MARY , KOBES SAYUKO , CHEN PENG , HSUEH WEN-CHI , NELSON ROBERT , BAIER LESLIE , KNOWLER WILLIAM C. , HANSON ROBERT L.
Published: June 2018

Inflammatory processes may contribute to type 2 diabetes (T2D), but there is limited information about genetic associations with circulating levels of inflammatory cytokines. We performed a genome-wide association study (GWAS) of cytokines in 1061 nondiabetic Southwestern Native Americans (500 women; 561 men).

Fasting serum concentrations of interleukin-6 (IL-6), plasminogen activator inhibitor-1 (PAI-1) and tumor necrosis factor-α (TNF-α) were measured using EMD Millipore assays. Genotypic data on 496,190 single-nucleotide polymorphisms (SNPs) with minor allele frequency >1% were derived from an Affymetrix Axiom array designed to capture common variation in Pima Indians. Data were normalized and analyzed for associations using a mixed model accounting for familial relationships, adjusted for age, sex and genetic principal components 1-5.

The strongest associations were on chromosome 3 near A4GNT with IL-6 levels (rs2724711 p=4.2×10-6), and on chromosome 7 with PAI-1 levels (rs1608488 (p=8.3×10-7). An association of genome-wide statistical significance (rs1862840; p=1.2×10-8) was observed on chromosome 16 near WWOX with TNF-α levels. Each copy of the G allele (frequency=0.12) confers a 0.41-SD increase in TNF-α levels. In a larger cohort of Southwestern Native Americans, rs1862840 was not associated with T2D (N=7,659; p=0.17). Among 3,983 nondiabetic subjects it did associate with 2 hour corrected insulin response (β=0.06-SD decrease per G allele copy, p=0.011). Among 300 normal glucose tolerant subjects it associated with the acute insulin response to intravenous glucose (β=20% decrease per G allele copy, p=0.045).

This study identified a novel association for TNF-α levels at WWOX (a gene previously implicated in T2D and insulin secretion in Han Chinese study populations) and suggests potential genetic links between inflammation and insulin secretion.

Disclosure

L.E. Wedekind: None. M. Walter: None. S. Kobes: None. P. Chen: None. W. Hsueh: None. R. Nelson: None. L. Baier: None. W.C. Knowler: None. R.L. Hanson: None.
Sleep Disturbances and Daytime Sleepiness in Pregnant Women with Diabetes
Authors: FACANHA CRISTINA , SOUSA FILIPE L. , FACANHA ARTHUR S. , FORTI ADRIANA C. , ROCHA HELLEN CRISTINA L. , PINHEIRO DE ARAUJO MARIANA SILTON , BRUIN VERALICE M.S.
Published: June 2018

Sleep disturbances in pregnancy are associated with depression, preterm delivery, hypertension, hyperglycemic disorders, and higher rate of cesarean as well as adverse perinatal outcomes. Diabetes is other prevalent condition in pregnancy related to these adverse outcomes, and insulin resistance is a common disruptor. As such, it is important to evaluate the quality of sleep and daytime somnolence in diabetic pregnancy.

Aim: Evaluate sleep quality and excessive daytime sleepiness in pregnancy complicated by diabetes mellitus. This is a cross-sectional study involving 115 pregnant women with diabetes (PWD), gestational age from 10 to 39 weeks. Sleep quality was assessed by the Pittsburgh Sleep Quality Index (PSQI-BR) and daytime sleepiness by the Epworth Sleepiness Scale (ESS). The study was approved by IPADE ethic board (1.801.860). Statistical analysis was performed using the software IBM SPSS.

Results: Poor sleep quality (PSQI>5) was observed in60,4% of PWD, which is higher them most reported rates for normal pregnancy (60,4% vs. 45% p: 0.01, CI99%. SEDOV, 2017) but similar to reported data on Brazilian overweight pregnant women (65,9%; RIBEIRO 2015). The mean PSQI score was 6.91. Short sleep duration was present in 35,8% (mean sleep duration: 7,2 hours/night). Sleep was disturbed by frequent urination in 66,1% of PWD. Daytime somnolence reported in 20.8% of PWD, and was not related with the quality of sleep. The higher number of parity was independently associated with poor sleep quality [p=0.03; 0R=1.74; CI=1.03-2.97]. The presence of obesity, hypertension, educational level, type of diabetes or treatment were non-influential to sleep quality or daytime sleepiness.

Conclusion: Poor sleep quality and excessive daytime sleepiness are frequent in PWD and a higher number of parity is independently associated with poor sleep quality. Given that sleep disturbances are related to adverse outcomes in PWD, we suggest that this group of patients should be the focus of therapeutic measures to improve sleep.

Disclosure

C. Facanha: None. F.L. Sousa: None. A.S. Facanha: None. A.C. Forti: None. H.L. Rocha: None. M. de Araujo: None. V.M.S. Bruin: None.
Serum Resistin Was Inversely Associated with Physical Activity in the C/C Genotype of SNP-420 in the General Japanese Population—The Toon Genome Study
Authors: KAWAMURA RYOICHI , TABARA YASUHARU , TAKATA YASUNORI , TAKAKADO MISAKI , MATSUSHITA YUMI , HADATE TOSHIMI , ONUMA HIROSHI , MARUYAMA KOUTATSU , TANIGAWA TAKESHI , SAITO ISAO , OSAWA HARUHIKO
Published: June 2018

In humans, resistin is predominantly expressed in monocytes and macrophages, and involved in insulin resistance, atherosclerosis, and cardiovascular diseases. We previously reported that the G/G genotype of single nucleotide polymorphism (SNP) at -420 (rs1862513) in the human resistin gene increased susceptibility to type 2 diabetes. Genetically, serum resistin was positively associated with the number of the G-allele at SNP-420 in the general Japanese population. Epigenetically, serum resistin was inversely associated with methylation of the C-allele at SNP-420. However, interaction between SNP-420 and environmental factors remains to be elucidated. The aim of this study was to investigate the association between serum resistin and physical activity, and the effect of SNP-420 on this association. The Toon Genome Study is a cohort study of Japanese community-dwelling subjects. A total of 1981 participants were cross-sectionally analyzed. Serum resistin was measured by ELISA. Physical activity was measured for 7 days using a waist-worn accelerometer (Lifecorder, Suzuken Co). Physical activity was also assessed using a questionnaire (the Japan Arteriosclerosis Longitudinal Study Physical Activity Questionnaire, JALSPAQ). Serum resistin was inversely associated with the quartile of moderate-intensity (3Mets or more) physical activity time (Q1 12.8, Q2 12.2, Q3 12.3, Q4 11.6 ng/ml; P for trend = 0.005; after adjustment for age, gender, and BMI). This inverse association was strongest in the C/C genotype of SNP-420. HOMA-IR was positively associated with serum resistin, and inversely associated with moderate-intensity physical activity time in the C/C genotype (Q1 1.51, Q2 1.52, Q3 1.36, Q4 1.32; P for trend = 0.01; after adjustment for age, gender, and BMI). Therefore, serum resistin and insulin resistance were inversely associated with physical activity in the C/C genotype of SNP-420 in the general Japanese population.

Disclosure

R. Kawamura: None. Y. Tabara: None. Y. Takata: None. M. Takakado: None. Y. Matsushita: None. T. Hadate: None. H. Onuma: None. K. Maruyama: None. T. Tanigawa: None. I. Saito: None. H. Osawa: None.
A Missense (c.T1424C:p.L475P) Mutation of ZYG11A Causes Maturity-Onset Diabetes of the Young
Authors: CHAROENSUK CHUTIMA , JUNGTRAKOON PRAPAPORN , TANGJITTIPOKIN WATIP , SUJJITJOON JATUPORN , PLENGVIDHYA NATTACHET , YENCHITSOMANUS PA-THAI
Published: June 2018

Maturity-onset diabetes of the young (MODY) is a monogenic form of diabetes inherited in an autosomal dominant mode. Approximately, 80% of Thai MODY families did not carry mutations of known MODY genes. This study aims, therefore, to identify an unknown causative gene of MODY in Thais. Exome sequencing was performed in DNA samples of two diabetic and one nondiabetic members of a MODY family with unknown genetic etiology. Only heterozygous non-synonymous variants with minor allele frequencies <0.01%, which were shared between the two diabetic but not existed in nondiabetic family members, were selected for segregation analysis. Among these, a missense mutation (NM_001004339, c.T1424C:p.L475P) in a gene encoding zyg-11 family member A (ZYG11A) showed complete segregation with diabetes in three generations. This mutation is not present in any database and not detected in nondiabetic controls (n = 400) as well as other MODY-X probands (n = 90). The amino acid residue L475 of ZYG11A is evolutionally conserved among many species. Several prediction software tools suggested a deleterious effect of the L475P mutation. ZYG11A acts as a target recruitment subunit of an E3 ubiquitin ligase complex, which plays an important role in degradation of cyclinB1- a regulatory protein involved in mitosis. Suppression of ZYG11A in 1.1B4 β-cell line by specific small interfering RNA (siRNA) significantly reduced cell proliferation (p = 0.001) while increased cyclinB1 protein levels (p = 0.001). Moreover, 1.1B4 cells overexpressing ZYG11A showed a significant increase in proliferation as compared to the control cells (p = 0.019) whereas this effect was not observed in the cells overexpressing the mutant ZYG11A (p = 0.659). Taken together, this finding demonstrates the crucial role of ZYG11A in β-cell cycle regulation. It also suggests heterozygous ZYG11A-L475P mutation as a cause of MODY in the Thai family studied.

Disclosure

C. Charoensuk: None. P. Jungtrakoon: None. W. Tangjittipokin: None. J. Sujjitjoon: None. N. Plengvidhya: None. P. Yenchitsomanus: None.
TNF-α in Combination with Palmitate Enhances IL-8 Production via TLR4-Dependent Signaling Pathway—Potential Relevance to Metabolic Inflammation
Authors: AHMAD RASHEED , AKHTER NADEEM , KOCHUMON SHIHAB P. , ALROUB AREEJ ABU , THOMAS REEBY S. , SINDHU SARDAR T.
Published: June 2018

Several chemokines produced by the adipose tissue in obesity are involved in metabolic inflammation. IL-8 or CXCL-8, a chemokine, is produced by immune cells and adipocytes and the increased levels of IL-8 are associated with several inflammatory disorders. Elevated IL-8 plasma levels have also been linked to insulin resistance (IR) and associated complications. It is still an open debate how does the obesity lead to an overwhelming production of IL-8. Not only the saturated free fatty acids and TNF-α are found to be elevated in obese humans but both have also been implicated with IR. Therefore, we sought to determine how these two obesity-associated factors could influence the production of IL-8 in human monocytic cells. Our data show that human monocytic cells/macrophages that were co-stimulated with palmitate and TNF-α synergistically increased IL-8 production. Of note, the synergistic production of IL-8 by TNF-α/palmitate was suppressed either by neutralizing TLR4 or by genetic silencing of TLR4. Moreover, inhibiting the clathrin-dependent endocytosis of TLR4 significantly suppressed the synergistic production of IL-8. This synergy between TNF-α and palmitate for IL-8 production was partly blocked by IRF3 knock down or TRIF inhibition. Both MyD88-deficient and MyD88-competent cells responded comparably to TNF-α/Palmitate co-stimulation. Interestingly, increased NF-κB/AP-1 activity was detected in the TNF-α/palmitate co-stimulated cells. Our human data show that increased adipose tissue TNF-α expression correlated positively with IL-8 expression (r=0.44, p=0.003). IL-8 also correlated positively with the fasting blood glucose (r=0.32, p=0.012), HBA1C (r=0.36, p=0.01 and HOMA-IR (r=0.30, p=0.05). These findings suggest that the cross-talk between saturated fatty acid palmitate and TNF-α signaling pathways is a key driver of the inflammatory response in obesity via an excessive production of IL-8.

Disclosure

R. Ahmad: None. N. Akhter: None. S.P. Kochumon: None. A. Abu Alroub: None. R.S. Thomas: None. S.T. Sindhu: None.
Pharmacological Inhibition of Neutrophil Elastase Attenuates Insulitis and Autoimmune Diabetes in Mice
Authors: LINGLING SHU , ZHONG LING , HOO RLC , XU AIMIN
Published: June 2018

Background: Type 1 diabetes is an autoimmune disease resulted from self-destruction of insulin-producing pancreatic beta-cells. However, the pathological pathways that trigger the autoimmune destruction remain poorly understood. Our previous studies demonstrated that increased circulating neutrophil elastase (NE) is closely associated with beta-cell autoimmunity in patients with T1D. Here we investigate the role and mechanism whereby NE participates in the pathogenesis of T1D, and to explore the therapeutic potential of the pharmacological NE inhibitors for autoimmune diabetes.

Method: Biochemical and immunological analysis were adopted to determine the dynamic change in the infiltration and activation of neutrophils in pancreas of NOD mice. NE-specific endogenous (pre-elafin/elafin) or pharmaceutical (sivelestat) inhibitors were supplemented into NOD mice to evaluate their effects on diabetes incidence and insultitis. Flow cytometry was employed to explore the effects of NE in mediating the crosstalk of immune cells.

Results: A dynamic change in the infiltration of neutrophils was observed in pancreatic islets of NOD mice, leading to a transient elevation of local NE activity. Supplementation of NE-specific inhibitors rectified the elevated pancreatic NE activity, alleviated insulitis and reduced the development of T1D in NOD mice. Inhibition of NE activity significantly attenuated the infiltration of pro-inflammatory macrophages into pancreas, reduced the production of inflammatory cytokines. In vitro studies showed that NE directly induced inflammatory responses in both min6 beta-cells and RAW264.7 macrophages through the activation of TLR4-IκB-NFκB pathway.

Discussion: NE-mediated crosstalk between beta-cell and macrophages forms a vicious feedback loop, resulting in the exaggerated beta-cell damage in NOD mice. Pharmacological inhibition of NE may represent a promising therapeutic strategy for treatment of autoimmune diabetes.

Disclosure

S. Lingling: None. L. Zhong: None. R. Hoo: None. A. Xu: None.
The MultiPepT1De Study—Examining the Safety of Peptide Immunotherapy Using Multiple Islet Antigens in Recent-Onset Type 1 Diabetes
Authors: LIU YUK-FUN , POWRIE JAKE K. , ARIF SEFINA , FOUNTOULAKIS NIKOLAOS , JOSHI MAMTA , SMITH EMMA L. , STRIMENOPOULOU FOTEINI , THOMSON MOIRA , PEAKMAN MARK
Published: June 2018

Background: Peptide immunotherapy (PIT) aims to modulate immune responses to specific antigens in order to restore immune tolerance. Recently, we reported outcomes of a phase 1b study of PIT at type 1 diabetes onset, showing safety, tolerability and changes in immune regulation pathways, following up to 12 doses of 10μg of a single proinsulin peptide that is naturally processed and presented by HLA-DR4 (DRB1*0401). Preclinical PIT studies show that multiple peptides in combination enhance tolerance induction.

Aims: To evaluate the safety and clinical effects of PIT using a mix of multiple islet autoantigen peptides in adults with recent-onset type 1 diabetes.

Methods: The MultiPepT1De study was a single centre, double-blind placebo-controlled, ascending dose, randomized trial recruiting adults aged 18-45 years within 4 years of diagnosis with HLA-DRB1*0401 genotype, positive islet autoantibodies and stimulated C-peptide >0.2pmol/mL. Study drug comprised of 6 naturally processed and presented, islet-derived peptides (MultiPepT1De). Participants were enrolled into 3 cohorts for monthly intradermal injections for 6 months. In each cohort, subjects were randomized to receive placebo (n=2) or MultiPepT1De (n=6) at 10µg (Cohort 1), 100µg (Cohort 2) or 500µg (Cohort 3) doses.

Results: Twenty-seven participants were randomized; 3 participants withdrew for non-clinical reasons and were replaced. There were no episodes of systemic hypersensitivity, anaphylaxis or serious adverse reactions linked to MultiPepT1De administration. Analysis of changes in stimulated C-peptide, HbA1c and insulin dose per kg showed no evidence of disease acceleration following dosing compared with placebo. Mild transient injection site erythema was observed in all cohorts, with no increase on repeated dosing.

Conclusions: Intradermal administration of multiple islet-derived peptides up to a cumulative dose of 3000µg, is well tolerated with no safety concerns.

Disclosure

Y. Liu: Other Relationship; Self; Novo Nordisk Limited. Research Support; Self; Bristol-Myers Squibb Company. J.K. Powrie: None. S. Arif: None. N. Fountoulakis: None. M. Joshi: None. E.L. Smith: Employee; Self; UCB, Inc. F. Strimenopoulou: Employee; Self; UCB, Inc. M. Thomson: Employee; Self; UCB, Inc. M. Peakman: Research Support; Self; Bristol-Myers Squibb Company. Advisory Panel; Self; Bristol-Myers Squibb Company. Consultant; Self; UCB, Inc.. Research Support; Self; UCB, Inc.. Advisory Panel; Self; T1D Exchange.
A Cut-Off Value for Anti-GAD Ab ELISA Test for Predicting Future Insulin Dependency in SPIDDM May Not Exist
Authors: OIKAWA YOICHI , KATSUKI TAKESHI , KAWAI TOSHIHIDE , SHIMADA AKIRA
Published: June 2018

We previously demonstrated that patients initially diagnosed with slowly progressive insulin-dependent (type 1) diabetes mellitus (SPIDDM) using a radioimmunoassay (RIA) test for anti-glutamic acid decarboxylase antibody (GADA) (GADA-RIA), who were also positive on a GADA enzyme-linked immunosorbent assay (ELISA) test (GADA-ELISA), showed a reduced insulin secretory capacity early in the SPIDDM disease process [Endocr J. 2017;64:163]. When we used GADA-RIA test, in SPIDDM with non-insulin dependent state, a GADA-RIA value ≥10 U/mL(GADA-RIA-High) was believed to indicate a higher risk for future insulin dependency than a value <10 U/mL(GADA-RIA-Low). In our previous study also, the insulin secretory capacity may be lower in “GADA-RIA-High and GADA-ELISA-positive SPIDDM patients” (A group) than those with “GADA-RIA-Low and GADA-ELISA-positive SPIDDM patients” (B group) early in the disease process, but this point was not yet confirmed. Therefore, we have now performed additional analysis to compare serum C-peptide levels in these two patient groups (group A vs. B). When restricted to a shorter disease duration, <11 years based on our previous study, serum C-peptide levels were 0.73 ± 0.61 ng/mL in group B (n = 9) and 0.68 ± 0.65 ng/mL in group A(n = 15). There was no significant difference between the two measures, but each of these two measurements was significantly lower than those for patients with type 2 diabetes (2.20 ± 0.85 ng/mL; n = 15) and those for “GADA-RIA-Low and GADA-ELISA-NEGATIVE SPIDDM patients” (2.60 ± 1.66 ng/mL; n = 6). These findings suggest that, regardless of their GADA-RIA titer, GADA-ELISA-positive patients with SPIDDM may show a reduced insulin secretory capacity early in the disease process.

In conclusion, unlike GADA-RIA test, a cut-off value for GADA-ELISA test for predicting future insulin dependency in SPIDDM may not exist.

Disclosure

Y. Oikawa: None. T. Katsuki: None. T. Kawai: None. A. Shimada: Speaker's Bureau; Self; Eli Lilly and Company, Sanofi-Aventis, Novo Nordisk A/S.
Alpha-1 Antitrypsin (AAT) Promotes Islets Graft Survival by Modulating Macrophage Activation
Authors: GOU WENYU , WANG JINGJING , KIM DO-SUNG , SONG LILI , STRANGE CHARLIE , ADAMS DAVID , MORGAN KATHERINE , WANG HONGJUN
Published: June 2018

Infiltrating macrophages can induce inflammation by secretion of proinflammatory cytokines and contribute to islet graft loss shortly after islet transplantation. In this study, we tested the mechanistic effects of AAT in mitigating macrophage mediated inflammation in an intrahepatic islet transplantation model in which human islets (2500 islet equivalent numbers) were transplanted into NOD-SCID mice that had been rendered diabetic (blood glucose >350mg/dl) by streptozotocin treatment. AAT (Prolastin-C, Grifols) was given to recipients intraperitoneally at 4 mg/mouse, every two days for 14 days, beginning before transplantation. In those recipients receiving AAT (n=29), 69.0% reached normoglycemia, compared to 35.7% in controls (n=28, p=0.002, logrank test) at 30 days post transplantation. Compared to mice treated with vehicle, AAT-treated mice had lower serum levels of interleukin 1 β (IL-1β), interferon- γ (IFN-γ) and interleukin 6 (IL-6), at day 1 post transplantation. To explore potential cell target(s) of AAT action, we analyzed activation of macrophages infiltrated into the liver after transplantation. Fewer M1 macrophages were observed in AAT-treated mice than in controls. To measure the effects of AAT on macrophage activation, we added AAT to Raw264.7 cells stimulated with IFN-γ in vitro. Expressions of M1 markers (iNOS, CD11c, TNFα, and IL-6) were dramatically decreased in AAT-treated cells compared to vehicle-treated controls. Co-culture of IFN-γ activated macrophages with islets led to islet cell apoptosis. Addition of AAT in the co-culture system reduced macrophage activation and protected islet cells from apoptosis.

In summary, AAT treatment significantly enhances human islet graft survival and function after transplantation into the liver of NOD-SCID mice, in part by suppression of macrophage activation and pro-inflammatory cytokine production.

Disclosure

W. Gou: None. J. Wang: None. D. Kim: None. L. Song: None. C. Strange: Research Support; Self; Grifols, CSL Behring, Shire. Consultant; Self; Shire. Stock/Shareholder; Self; Abeona. Consultant; Self; CSL Behring. Research Support; Self; Adverum. Consultant; Self; Adverum. Research Support; Self; MatRx. D. Adams: None. K. Morgan: None. H. Wang: None.
Protective Effect of NecroX-7 on Islets against Oxidative Stress and Inflammation
Authors: KWON YOUNGSANG , LEE HAN SIN , KIM HYUNJIN , KIM GYURI , JIN SANG-MAN , LEE MOON-KYU , KIM JAE HYEON
Published: June 2018

Oxidative stress generated during islet isolation, culture, and transplantation causes islet cell damage mediated by hypoxia, reactive oxygen species (ROS) and inflammatory responses. We investigated the protective effects of NecroX-7, a novel ROS scavenger, during isolation and/or culture of islets. For ex vivo studies, islets from heterozygote human islet amyloid polypeptide (hIAPP+/-) mice and C57BL/6J mice were isolated by collagenase with and without supplementation with 20 uM of NecroX-7. Supplementation with NecroX-7 provided markedly increased islet viability and ATP contents, and attenuated ROS, transcription of c-Jun N-terminal kinases, high mobility group box-1 (HMGB1) and proinflammatory cytokines including interleukin-1beta (IL-1b), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-a). Transplantation of islets, which is derived from islet isolation method using supplementation with NecroX-7, into syngeneic subrenal subcapsular of hIAPP+/- mice also improved post-transplant blood glucose levels. Also, NecroX-7 protected RINm5F cells from t-BHP-induced and serum deprivation induced toxicity in in vitro culture by dose-dependent manner from 0.1 uM to 20 uM. Supplementation of medium with NecroX-7 during serum-deprived culture condition also improved impaired viability and serum deprivation-induced ROS in islets from hIAPP+/- mice by dose-dependent manner from 0.1 uM to 20 uM. These findings suggest that NecroX-7 supplementation during the islet isolation and/or culture process suppressed inflammatory responses and ROS induced by serum deprivation and provides a potential benefit to improve post-transplantation outcomes.

Disclosure

Y. Kwon: None. H. Lee: None. H. Kim: None. G. Kim: None. S. Jin: None. M. Lee: None. J. Kim: None.
Synergistic Effect of Islet Magnetic Resonance Imaging Using Direct Linking of Ferumoxytol to Surface of PEGylated Islet
Authors: LEE HAN SIN , KWON YOUNGSANG , KIM HYUNJIN , HAQUE MUHAMMAD R. , KIM GYURI , JIN SANG-MAN , LEE MOON-KYU , BYUN YOUNGRO , KIM JAE HYEON
Published: June 2018

Purpose: We previously reported the feasibility of islet magnetic resonance imaging (MRI) using ferumoxytol, which is the only clinically-available ultrasmall superparamagnetic iron oxide. However, the labeling efficacy was not sufficient for its clinical use in islet transplantation. We aimed to evaluate the feasibility of islet MRI using direct linking of ferumoxytol to islet surface through PEGylation.

Method: Islets were labeled by surface modification with up to 4% PEG-heparin-ferumoxytol. We compared islet function and viability of control islets and ferumoxytol-heparin-PEGylated islets. Efficacy of labeling with ferumoxytol-heparin-PEGylation and ferumoxytol alone was assessed in both ex vivo and in vivo models.

Results: Labeling of islets with up to 2% PEG-heparin-ferumoxytol did not derange ex vivo islet viability and function. The T2* relaxation time was optimal when islets were labeled with 1 to 4% PEG-heparin-ferumoxytol. The labeling intensity in the ex vivo MRI of ferumoxytol-heparin-PEGylated islets was stronger than islets labeled with ferumoxytol alone. In syngeneic renal subcapsular islet transplantation, labeling with ferumoxytol-heparin-PEGylation showed better in vivo labeling efficacy than that of islets labeled with ferumoxytol alone. After labeling with ferumoxytol-heparin-PEGylation, there was a correlation between the total area of visualized islets and the transplanted islet mass in syngeneic mouse intraportal islet transplantation, and the visibility was also confirmed in the preliminary analysis of non-human primate intraportal islet transplantation model.

Conclusion: Direct linking of ferumoxytol to islet surface through PEGylation improved the labeling efficacy and it could be used for the islet MRI in clinical islet transplantation.

Disclosure

H. Lee: None. Y. Kwon: None. H. Kim: None. M.R. Haque: None. G. Kim: None. S. Jin: None. M. Lee: None. Y. Byun: None. J. Kim: None.
Intraperitoneally Administered Neo-islets, Aggregates of Mesenchymal Stem Cells, and Cultured Islet Cells Improve Glycemic Control of Spontaneously Occurring Insulin-Dependent Diabetes Mellitus in Pet Dogs—A Pilot Study, INAD 012-776
Authors: WESTENFELDER CHRISTOF , GOOCH ANNA , LOY SON NATASHA K. , AVILA NICOLE , FISCHER JULIE R.
Published: June 2018

The clinical need for novel technologies that effectively treat patients with T1DM and render them insulin-independent is great. Endogenous insulin replacement by pancreas or islet transplants is currently the only treatment that can achieve insulin-independence and provide significant end organ protection in patients with autoimmune-mediated T1DM. However, the shortage of pancreas donors and the need for repeated islet transplants, requiring up to five donors each, continue to limit the availability of these expensive therapies. Both transplant modalities depend on the permanent use of potentially toxic antirejection drugs. We previously reported that allogeneic, i.p. administered “Neo-Islets” (NIs), composed of cultured islet cells coaggregated with high numbers of immunoprotective and cytoprotective Adipose Derived Stem Cells (ASCs) reestablished normoglycemia in autoimmune T1DM NOD mice without the use of encapsulation devices or immunosuppressive agents. With FDA guidance, we are currently testing this NI technology in a pilot study of insulin-dependent, spontaneously diabetic pet dogs by the i.p. administration of 2x10e5 NIs/kg bw to metabolically controlled (blood glucose, triglycerides) dogs under light anesthesia and ultrasound guidance. Dogs are continued on insulin as needed and are to be followed for 3 years for adverse events, glycemic control, insulin need. Two dogs have been treated. One has been followed for 6 months, the other for 3 months. While decreasingly insulin dependent with significantly improved glycemic control, and no IgG response to the allogeneic NIs, no adverse events have been observed.

In conclusion, these data demonstrate that this novel technology is safe, feasible and holds promise for the treatment of dogs with insulin dependent DM, but that further protocol/dosing optimization is required.

Disclosure

C. Westenfelder: Consultant; Self; SymbioCellTech, LLC. A. Gooch: Other Relationship; Self; SymbioCellTech, LLC. N.K. Loy Son: None. N. Avila: None. J.R. Fischer: None.
Adipocyte EGFL6 Expression from Subcutaneous Adipose Tissue Alters Glucose Homeostasis and Affects Human Obesity
Authors: BRADLEY DAVID , YIN ZHENG , LIU JOEY Z. , BLASZCZAK ALECIA M. , WONG STEPHEN T. , HSUEH WILLA
Published: June 2018

Recent data suggests that the adipose tissue (AT) extracellular matrix (ECM) may be a key player in the pathogenesis of obesity-related T2D. HFD feeding to collagen VI knockout mice reduces AT fibrosis (a hallmark of abnormal ECM) and improves insulin sensitivity despite an expansion of adipocyte volume. Epidermal growth factor-like-domain, multiple 6 (EGFL6) is a secreted protein best characterized for its role in tumor endothelial cell angiogenesis. Although previous studies have shown increased EGFL6 expression in whole fat of obese and insulin resistant subjects, its precise role in metabolic disease, the subtypes of cells within AT that overexpress EGFL6, and the mechanism(s) by which AT EGFL6 could modulate insulin sensitivity are entirely unknown. We obtained SAT from twenty-five lean (BMI 22.8±0.3 kg/m2) and twenty-nine obese (BMI 50.2±1.6 kg/m2) human subjects. Adipocytes and stromal vascular fraction were isolated for gene expression and co-culture experiments. We found that both adipocyte (37.21±7.55 vs. 7.44±3.67,p=0.001) and macrophage (4.69±0.79 vs. 1.15±0.45,p=0.024) EGFL6 expression was highly upregulated in human obesity; yet, only adipocyte-derived EGFL6 related significantly to insulin sensitivity (r=+0.775,p<0.001), even independent of BMI (Beta=+0.676,p<0.001). We also noted a marked difference in adipocyte EGFL6 (13.0±4.0 vs. 45.1±17.3,p=0.043) between those subjects who were insulin sensitive (HOMA-IR<3) compared to those who were insulin resistant (HOMA-IR≥3). Mechanistically, we further show that recombinant EGFL6 can directly disrupt insulin signaling in cultured adipocytes, hepatocytes, and skeletal muscle cells. Our findings linking adipocyte EGFL6 with systemic IR may lead to novel pathways involving adipocyte-derived EGFL6 and additional therapeutic targets for human T2D.

Disclosure

D. Bradley: None. Z. Yin: None. J.Z. Liu: None. A.M. Blaszczak: None. S.T. Wong: None. W. Hsueh: Advisory Panel; Self; Merck & Co., Inc., AstraZeneca, Novo Nordisk Inc..
FSP27-Rab18-Vimentin Regulates Lipid Droplet Trafficking and Insulin Signaling in Adipocytes
Authors: PURI VISHWAJEET , BANERJEE SAYANI , JASH SUKANTA
Published: June 2018

Increased free fatty acid flux from white adipocytes exacerbates insulin resistance and increases the risk of its progression to type 2 diabetes. Distinct intracellular organelles called lipid droplets (LDs) in adipocytes play a key role in storing fatty acids (FAs) as triglycerides (TGs), hence protecting against FA-induced lipotoxicity. A highly coordinated cascade of LD-associated proteins and lipases is known to regulate the catabolism of stored TGs in LDs, but the molecular components of LDs that determine their dynamics and TG storing capacity in adipocytes remains understudied. Previously, we showed that Fat specific protein 27 (FSP27) is a lipid droplet associated protein that is positively associated with insulin sensitivity in humans, and plays a crucial role in lipid droplet dynamics by clustering and then fusion of LDs. The present investigation was designed to identify the molecular mechanism through which FSP27 mediates LD trafficking to regulate the initial step of LD clustering—a prerequisite of LD fusion. We found that a direct interaction between FSP27, Vimentin and Rab18, and FSP27-mediated activation of Rab18 GTPase activity is important for lipid droplet trafficking. Our results reveal a complex bi-directional cross-talk that takes place between LD-associated proteins and cytoskeletal elements that maintain lipid droplet dynamics in a highly organized manner. Our study provides a new insight in the regulation of LD dynamics and insulin signaling in adipocytes.

Disclosure

V. Puri: None. S. Banerjee: None. S. Jash: None.
Silencing Mediator of Retinoid and Thyroid Hormone Receptors (SMRT)—A Link between Adipocytes, Inflammation, and Glucose Intolerance
Authors: KAHN JONATHAN , GODDI ANNA A. , COHEN RONALD N.
Published: June 2018

Silencing Mediator of Retinoid and Thyroid Hormone Receptors (SMRT) is a nuclear corepressor, which regulates the transcriptional activity of metabolically essential transcription factors. While SMRT has been shown to modulate adipocyte function, in vivo experiments utilizing knock-out (KO) models have led to conflicting results. To more rigorously define the role of SMRT in the adipocyte, we generated adipocyte-specific SMRT KO (adSMRT KO) mice by crossing adiponectin-Cre mice with floxed SMRT mice on a C57/BL6 background. When adSMRT KO mice are challenged with a 45% high-fat diet, we observe 20% increase in glucose intolerance (p=0.006) compared to wild type (WT) counterparts. Additionally, RNA-Seq data of adipose tissue from these mice indicate a dramatic up-regulation of inflammatory gene expression. To further characterize the pro-inflammatory phenotype, we utilized flow cytometry to identify infiltration of specific populations of adipose tissue inflammatory cells. We found that overall macrophage infiltration in the adipose tissue of KO mice increased two-fold (p=0.026), with anti-inflammatory M2 macrophages infiltrating in significantly lower proportions (p=0.017), indicating a higher ratio of M1:M2 cells in the KO mice compared to WT. In contrast, body weight and total fat mass were not altered. These data suggest a role for SMRT in the cross-talk between adipocytes and pro-inflammatory macrophages for the regulation of systemic glucose tolerance, distinct from the development of obesity. SMRT therefore integrates metabolic and inflammatory signals to maintain physiological homeostasis.

Disclosure

J. Kahn: None. A.A. Goddi: None. R.N. Cohen: None.
Remodeling of Myocardial Metabolism by Cardiac Mitochondrial Akt Signaling through Modulation of Pyruvate Dehydrogenase Complex
Authors: CHEN YU-HAN , CHEN YUMAY , TA ALBERT , LEE HSIAOCHEN , LIN HUGO YOU-HSIEN , WANG PING H.
Published: June 2018

Mitochondria play a critical role in the regulation of myocardial metabolism and function. Diminished insulin-activated Akt translocation into mitochondria matrix contributed to the development of diabetic cardiomyopathy. However, its mechanisms of actions are not fully understood. Using an inducible heart specific transgenic mice harboring a mitochondria-targeting dominant Akt (CAMDAKT), we have observed 50% attenuation of pyruvate dehydrogenase activity in mitochondria. Proteomic study revealed insulin injection promoted formation of pyruvate dehydrogenase supercomplex in the control mice, however, insulin-stimulated formation of pyruvate dehydrogenase supercomplex was inhibited in the CAMDAKT mice. We next used a novel small molecular inhibitor (SMI) to disrupt binding of Akt to the E3 subunit of pyruvate dehydrogenase to dissect how active Akt interacted and modulated pyruvate dehydrogenase complex (PDC). Insulin stimulation of PDC activity and complex formation were inhibited by the SMI. Acetyl-CoA, which bridges PDC to the TCA cycle, was significantly reduced by 40% in the cardiac mitochondria of CAMDAKT mice. Interestingly, this was accompanied by reduced NAD+ and NADH levels (40-50% reduction) and NAD+/NADH ratio (30-40% reduction). Therefore the redox state was compromised. In CAMDAKT mice, cardiac mitochondria underwent adaptive changes once mitochondrial Akt became inhibited, with biphasic alteration of mitochondria abundance (up and down). The expression of PPARγ and PGC1α, showed a similar pattern of changes, suggesting compensatory mitochondria biogenesis.

In summary, mitochondrial Akt action in the heart is at least partially mediated through its direct interaction with the E3 subunit of PDC, and inhibition of this pathway led to metabolic remodeling and compensatory mitochondria biogenesis. These data shed new light into the mechanism of mitochondrial Akt actions and how it modulated cardiac metabolism.

Disclosure

Y. Chen: None. Y. Chen: None. A. Ta: None. H. Lee: None. H. Lin: None. P.H. Wang: None.