bluuebunny/crossref_metadata_2025 huggingface dataset:
Insulin Resistance and Weight Loss in Obese Women with Polycystic Ovary Syndrome Treated with Liraglutide—Single Institution Experience
Authors: CRAWFORD ANDREW , COONEY LAURA G. , JOCHYM NICOLE , DOKRAS ANUJA , AMARO ANASTASSIA
Published: June 2018

Background: Insulin resistance contributes to the pathogenesis of polycystic ovary syndrome (PCOS) in obese women, but its predictive value for weight loss with liraglutide treatment is unknown.

Methods: Retrospective cohort study of 16 obese (BMI ≥30 kg/m2), nondiabetic women aged 18-42 with PCOS who were seen in a weight loss clinic between November 2014 and November 2016 and prescribed liraglutide (either 1.8 mg/day or 3.0 mg/day based on insurance coverage) and metformin. Maximum weight loss achieved from baseline until 2 to 13 months post-liraglutide initiation was compared. Spearman’s rank correlation was used to establish the relationship between pre-treatment Homeostatic Model for Insulin Resistance (HOMA-IR) and % weight loss. The University of Pennsylvania Institutional Review Board approved the study.

Results: Overall median weight loss was 4.4 kg (-3.4% of initial weight). Median weight loss was -4.7 kg (-3.7%) in women treated with 3.0 mg daily (n=6) and -2.5 kg (-2.0%) in women treated with 1.8 mg (n=10). 6/16 (37.5%) of women experienced >5% weight loss: 2/6 women treated with 3.0 mg daily dosing, and 4/10 women treated with 1.8 mg daily. HOMA-IR at baseline was not associated strongly with % weight loss achieved for all subjects (rs= -0.31, p=0.24), or when stratified to examine those subjects receiving 3.0 mg daily (rs= -0.20; p=0.70) or 1.8 mg daily (rs= -0.36, p=0.31).

Conclusions: In this small study, our findings suggest that insulin resistance at baseline does not strongly correlate with maximum weight loss achieved using liraglutide. Furthermore, our study suggests that liraglutide is an effective tool for weight loss. Overall, 37.5% of women achieved >5% weight loss from baseline, meeting the FDA’s categorical efficacy target for weight loss medications.

Disclosure

A. Crawford: None. L.G. Cooney: None. N. Jochym: None. A. Dokras: Consultant; Self; Fractyl Laboratories, Inc. A. Amaro: Consultant; Self; Novo Nordisk Inc., Andrew Technologies, LLC.
Gut Microbiota as a Potential Target for Treatment of Polycystic Ovary Syndrome
Authors: ZHANG BINGJIE , SHEN SHANMEI , BI YAN , ZHU DALONG
Published: June 2018

Objective: Emerging evidence has linked the gut microbiota with human metabolic diseases. This study is aimed at investigating the role of gut microbiota and the potential effect of gut microbiota modulation in polycystic ovary syndrome (PCOS).

Methods: We performed an observational study in a cohort of reproductive women with and without PCOS. Subjects with previous history of gastrointestinal surgery or diseases (such as inflammatory bowel disease), taking antibiotics within 3 months prior to the study were excluded. Diagnosis of PCOS was done according to Rotterdam criteria. A total of 78 women were finally included, information regarding their clinical history, symptoms, anthropometric measurements and laboratory data were collected. Fecal bacteria profiling was conducted using 16S rRNA sequencing method. Individuals with PCOS were then randomized to metformin or probiotics (containing Lactobacillus and Bifidobacterium) treatment for 3 months.

Results: We identified that women with PCOS had lower fecal bacteria gene counts, lower bacterial diversity and higher β diversity, indicating lower gene richness and a more heterogeneous gut microbiota community structure. In addition, serum lipid profiles (including total cholesterol, HDL-cholesterol), androgen levels and indexes of insulin resistance (HOMA-IR, Matsuda insulin sensitivity index) were closely correlated with Prevotellaceae, Ruminococcaceae and Lachnospiraceae, which are dominant bacteria in energy metabolism. Furthermore, treating PCOS patients with metformin or probiotics both improved their menstruation cycles, decreased androgen levels and alleviated insulin resistance. Meanwhile, there were significant correlations between changes in metabolic parameters and shifted bacterial populations.

Conclusion: Our findings identify the gut microbiota dysbiosis in PCOS and suggest that it may be possible to intervene in PCOS by targeting the gut microbiota.

Disclosure

B. Zhang: None. S. Shen: None. Y. Bi: None. D. Zhu: None.
Serum Retinol Binding Protein 4 Was Positively Associated with Insulin Resistance and Decreased after Laparoscopic Sleeve Gastrectomy in Chinese Obese Patients
Authors: WANG XINGCHUN , LI LIANG , ZHU CUILING , GAO JINGYANG , QU SHEN
Published: June 2018

Retinol binding protein 4 (RBP4) is implicated in the regulation of energy metabolism and insulin sensitivity. Laparoscopic sleeve gastrectomy (LSG) has been proven effectively in treating T2DM and other obesity-related comorbidities. We aimed to investigate the association between RBP4 with insulin resistance and the change of RPB4 levels after laparoscopic sleeve gastrectomy (LSG) in Chinese obese patients. Eighty-two obese patients (34 males and 48 females) were enrolled and 30 subjects (10 males and 20 females) underwent LSG in this study. Anthropometric and glucose metabolic parameters were measured. Serum RBP4 levels were detected by enzyme-link immunosorbent assay. In all subjects, serum RBP4 levels were positively associated with BMI, OGTT blood glucose at 0, 30, 120, 180mins, fasting insulin, fasting C-peptide and HOMA-IR at baseline (all P<0.05). Moreover, HOMA-IR was significantly decreased at 3 and 6 months after LSG (all P<0.001). Intriguingly, serum RBP4 levels were also reduced from baseline to 3 and 6 months after LSG (from 43.20±14.27mg/l to 33.00±12.35mg/l to 31.87±9.90mg/l, all P<0.01).

In conclusion, RBP4 is closely associated with glucose metabolism and insulin resistance in Chinese obese patients. The decreased RBP4 levels may partly account for the improved insulin sensitivity in obese subjects after LSG.

Disclosure

X. Wang: None. L. Li: None. C. Zhu: None. J. Gao: None. S. Qu: None.
Association between IGF1/IGFBP3 with Lipid Metabolism and Chronic Low-Grade Inflammation and Alteration after Laparoscopic Sleeve Gastrectomy in Chinese Obese Subjects
Authors: LI LIANG , WANG XINGCHUN , GAO JINGYANG , QU SHEN
Published: June 2018

Some studies have demonstrated the disturbance of IGF1/IGFBP3 axis in endocrine-metabolic diseases. We aimed to investigate the association between IGF1/IGFBP3 with glucose-lipid metabolism and chronic low-grade inflammation and the changes after laparoscopic sleeve gastrectomy (LSG) in Chinese obese patients. Thirty-six obese subjects were enrolled. Anthropometric data, routine laboratory biochemical parameters and IGF1/IGFBP3 levels were evaluated at baseline and 3 months after LSG. Serum total IGF1 and IGFBP3 levels were measured by enzyme-link immunosorbent assay and molar ratio of IGF1/IGFBP3 is represented as free IGF1 levels. In all subjects, total IGF1 was positively correlated with HDL-c (P<0.05), negatively with weight, BMI, waistline, hipline, waist/hip ratio, C-reactive protein (CRP) and tumor necrosis factor-α (TNF-α) (P<0.or P<0.01). Free IGF1 was negatively correlated with hipline, triglyceride (TG), CRP and TNF-α (P<0.or P<0.01). IGFBP3 was positively correlated with TG and free fatty acids (FFA) (P<0.or P<0.01). Both total IGF1 and IGFBP3 levels were significantly decreased after LSG (from 166.54±64.44ng/ml to 143.02±69.09ng/ml, P=0.048 and from 2556.54±756.99ng/ml to 2051.89±618.61ng/ml, P<0.001, respectively, and more reduction in IGFBP3 levels of 19.74%), while an increased tendency of free IGF1 levels was observed. In addition, changes of total IGF1/free IGF1 were negatively related to changes of CRP and TNF-α (P<0.or P<0.01), and change of IGFBP3 was positively related to change of FFA (P<0.05).

In conclusion, the IGF1/IGFBP3 axis is closely associated with lipid metabolism and chronic low-grade inflammation in Chinese obese patients. Decreased IGFBP3 and increased free IGF1 may partly account for the improved lipid metabolic and chronic low-grade inflammatory profiles after LSG.

Disclosure

L. Li: None. X. Wang: None. J. Gao: None. S. Qu: None.
Glycemic and Metabolic Effects of Liraglutide as Additive Treatment to Different Therapeutic Regimens in Type 2 Diabetes Patients Followed in Primary Care Clinics
Authors: ALHARBI TURKI , TOURKMANI AYLA , ALTWAIJRI SULAIMAN , KOFI MOSTAFA
Published: June 2018

Background: Liraglutide improved metabolic derangement in type 2 diabetes (T2D). The study aimed to measure changes in glycemic and metabolic parameters after addition of liraglutide in obese insulin-resistant T2D patients

Methods: liraglutide was added to 45 Adult T2D patients, who were on 4 regimens as follows:18 on Sulphonylurea(SU), 9 on SU and Basal insulin, 3 on basal insulin and 15 on multiple daily insulin injections(MDIs). Five patients had history of cardiovascular diseases (CVD).The cohort was followed for 6 months and Liraglutide dose 0.6 escalated to 1.2 mg based on patient tolerance. Glycemic, metabolic and anthropometric parameters were measured every 3 months

Results: Mean age and T2D duration were 51.5±12.9 and 11.6±1.6 years, respectively. Body weight significantly reduced from 98.7kg to 96.0kg, p< 0.001, with a mean difference of 3.39kg ± 5.6kg. Systolic blood pressure reduced from 131.7mmHg to 126.6mmHg (Diff. 6.1mmHg), p = 0.015 while Diastolic blood pressure reduced from 73.3mmHg to 68.6mmHg (Diff. 4.7mmHg), p = 0.026. Most HbA1c reduction observed in the liraglutide 1.2 mg dose group with changes from 9.8% to 8.2% (Diff. 1.6%); and also the most weight reduction observed in the 1.2 mg liraglutide dose group with changes from 103.3kg to 99.7kg (Diff. 3.6kg) compared to 0.6 and 1.8 mg dose groups. HbA1c reduction was largest in MDIs group from 10.2 to 8.6% (Diff. 1.6%). Meanwhile, the most weight reduction noticed in the treatment group on SU and Basal insulin from 97.2kg to 92.8kg (Diff. 4.4kg). Interestingly, patients with CVD had improved HbA1c with changes from 10.62% to 9.77% (Diff. 0.85%) and weight from 86.96kg to 85.72kg (Diff. 1.24kg) without evident hypoglycemia.

Conclusions: Addition of liraglutide to different T2D therapeutic regimen in obese Insulin- resistant patients showed significant improvement in HbA1c, weight, and Blood pressure. These results support the use in primary care clinics.

Disclosure

T. Alharbi: None. A. Tourkmani: None. S. Altwaijri: None. M. Kofi: None.
Obesity and Liver Function in a Group of Children Visiting a Pediatric Clinic in South Texas
Authors: QUINTANA FERNANDO G. , CADENA JULIA A. , CERVANTES-GONZALEZ FRANCISCO J.
Published: June 2018

It has been determined that there is a relationship between obesity and liver function. Purpose: the aim of this study is to analyze the relationship between obesity and liver function in a group of children.

Methods: the study was based on data obtained from 692 boys and 675 girls aged 0 to 15 years that visited the clinic in South Texas from 2000 to 2003. Children were classified according with the CDC BMI-for-age growth charts. Patients were classified also by age as stay home (SH) 0.5 to 3.99 years, elementary school (ES) 4 to to 11.99 years, Middle school (MS) 12 to 14.99 years and High school (HS) more than 15 years. Liver function was measured with the following enzymes alanine amino transferase (ALT), serum aspartate amino transferase (AST), and alkaline phosphatase (ALP). Analysis of variance was used for the analysis.

Results: there was a difference between obese and normal children for ALT (n = 45.18±0.83 IU/L vs. 38.87±1.09 IU/L, p<0.05). There were no differences for ALP or AST between obese and normal children. AST levels were higher in boys when compared with girls (n=691, 30.84±0.479 IU/L vs. n=675, 27.91±0.49 IU/L, p<0.05). ALT levels were higher in boys when compared with girls (n=691, 45.29± IU/L, n= 675, 39.30±0.66 IU/L, P<0.05). There was no difference for levels of ALP between boys and girls. There was a decrease of ALP levels with age when children are classified as SH, ES, MS, and HS (n = 288, 327.29±27.16 IU/L; n=791, 291.36±2.69 IU/L; n=151, 261.27±9.17 IU/L; n=134, 139.73±6.30 IU/L; p<0.05). There was a decrease of AST levels with age when children are classified as SH, ES, MS, and HS (n=288, 35.66±0.53 IU/L; n=791, 29.25±0.45 IU/L; n=151, 25.40±1.23 IU/L, n=134, 29.91±0.87, p<0.05).

Conclusions: obesity represents an important factor that affects ALT levels. ALT is directly related with BMI percentile. Boys have higher levels of liver enzymes than girls. As children attend school and leave home ALP and AST levels decrease.

The research was approved by TAMIU IRB with ID number #2012-01-13.

Disclosure

F.G. Quintana: None. J.A. Cadena: None. F.J. Cervantes-Gonzalez: None.
Assessment and Expansion of Models for 13C-Metabolic Flux Analysis (MFA) In Vivo
Authors: HASENOUR CLINTON , RAHIM MOHSIN , YOUNG JAMEY
Published: June 2018

Isotope-based MFA is a valuable tool to determine rates of biochemical reactions in vivo, which cannot be assessed by other approaches. Its application in biomedicine may help to uncover metabolic disease determinants in diabetes and obesity. However, various simplifying assumptions have been applied to constrain models used for in vivo MFA studies. These constraints are approximate, and their validity may change with varying experimental conditions. Here we used our flexible flux modeling platform (INCA) to test and eliminate several assumptions imposed on prior estimates of liver gluconeogenesis, anaplerosis, and citric acid cycle (CAC) fluxes in overnight fasted, conscious mice. Male C57Bl/6J mice were given a primed, continuous infusion (160µmol/kg and 40µmol/kg/minutes) of [13C3]lactate intravenously for ∼2 hour. 13C-enrichment of plasma and liver metabolites obtained at the end of the study were measured by GC-MS and used to regress liver metabolic fluxes using several different models of in vivo metabolism. In particular, we developed models to assess (1) the extent of reversibility of 4C reactions of the CAC, (2) the impact of pyruvate-decarboxylation, and (3) the influence of secondary isotope recycling on liver flux estimates. Confidence intervals were computed a posteriori to assess uncertainties of individual flux estimates. Glucose production originated largely from PEP. Cataplerosis (PEPCK flux) was ∼3.9-4.5 fold higher than citrate synthase, with ∼56-67% recycling back to the CAC through pyruvate. The modeling of pyruvate decarboxylation increased the relative ratio of gluconeogenesis to citrate synthase flux by ∼18-32% but also reduced the precision of flux estimates. Interestingly, Cori cycling reduced pyruvate kinase flux ∼16% relative to cataplerosis. Thus, interorgan crosstalk with the liver can be estimated with fewer assumptions in vivo, providing a potential path for understanding metabolic disease with greater consideration of whole-body physiology.

Disclosure

C. Hasenour: None. M. Rahim: None. J. Young: None.
Changes in Carbohydrate Metabolism are Related to Gut Microbiota Modification after H. Pylori Eradication
Authors: CORNEJO-PAREJA ISABEL , MARTIN-NUÑEZ GRACIA MARÍA , MAR ROCA-RODRIGUEZ M. , MORENO-INDIAS ISABEL , VINUELA LAURA , COIN-ARANGÜEZ LETICIA , MANCHA-DOBLAS ISABEL , TINAHONES FRANCISCO
Published: June 2018

Introduction: Antibiotic treatments cause changes in gut microbiota. Dysbiosis of gut microbiota has been linked to metabolic diseases such as type 2 diabetes mellitus (T2DM). Objective: To evaluate whether changes in gut microbiota due to antibiotic treatment in patients colonized by H. pylori, could be related to improvements in carbohydrate metabolism.

Material and Method: A prospective case-control study were performed. Clinical data, carbohydrate metabolism, and microbiota composition in feces (determined by 16S rRNA gene (V3-V4) sequencing using the Illumina Miseq) before and 2 months after eradication treatment were analyzed.

Results: We studied 40 cases and 20 controls (60% women, respectively). Average age was 47.0 ± 2 vs. 43.6 ± 2.7 years old. 70% vs. 60% had family history of digestive disorders and 42.5% vs. 40% clinical history of gastrointestinal disease. After antibiotics, we found significant changes in gut microbiota profile at phylum, family, genus and species levels. The Chao and Shannon indices showed a decreased in bacterial richness and diversity in patients (pre and post H. pylori eradication) compared to controls. We observed an improvement in glucose homeostasis in cases 2 month after H. pylori eradication treatment, with decrease in HbA1c (p=0.014), 60’ (p=0.018) and 120’ (p=0.019) glucose post OGTT. Changes in Rikenellaceae, Butyricimonas, E. biforme, B. fragilis, and Megamonas were inversely associated with changes in glucose level and Hb1Ac in patients which got H. pylori eradication.

Conclusion: H. pylori eradication treatment causes alteration in human gut microbiota. The glucose homeostasis improvement found after antibiotic treatment was related to changes in microbial community, particularly in Megamonas and Butyricimonas genus, Rikenellaceae family and E. biforme and B. fragilis species. SCFA-producing and glucose remover bacterias could play an important role in these associations.

Disclosure

I. Cornejo-Pareja: None. G. Martin-Nuñez: None. M. Roca-Rodriguez: None. I. Moreno-Indias: None. L. Vinuela: None. L. Coin-Arangüez: None. I. Mancha-Doblas: None. F. Tinahones: None.
Clinical Utility of the T1D Genetic Risk Score—Examples from the U.S. Monogenic Diabetes Registry
Authors: SANYOURA MAY , LETOURNEAU LISA R. , NAYLOR ROCHELLE N. , PHILIPSON LOUIS H. , WEEDON MICHAEL N. , ORAM RICHARD A. , GREELEY SIRI ATMA W.
Published: June 2018

Monogenic forms of diabetes are often difficult to distinguish from type 1 (T1D) and type 2 diabetes due to overlapping clinical features. As a result, a growing majority of patients remain misdiagnosed and often inappropriately treated. We used a T1D genetic risk score (T1D-GRS), generated from 10 T1D-associated common genetic variants, to help identify patients with a low T1D-GRS, and thus a potential monogenic etiology. We genotyped 798 probands from the U.S. Monogenic Diabetes Registry: 37% (n=293) had a known monogenic cause and 63% (n=505) had an unknown cause or were not exhaustively sequenced. To improve the discriminatory power of the T1D-GRS, participants were categorized using a T1D-GRS equivalent to the 5th, 25th, 50th, 75th, and 95th centile of the UK T1D cohort. 45% (132) of the registry participants with a confirmed monogenic cause had a T1D-GRS below the 5th percentile, 35% (103) had a T1D-GRS between the 5-25th percentile, 12% (35) between the 25-50th percentile, 5% (15) between the 50-75th percentile, and 2% (7) between the 75-95th percentile. Of the participants with an unknown cause, 24% (n=108) had a T1D-GRS below the 5th percentile, 30% (n=137) had a T1D-GRS between the 5-25th percentile, 16% (72) between the 25-50th percentile, 15% (67) between the 50-75th percentile, and 15% (69) between the 75-95th percentile. We sequenced a total of 124 participants with a T1D-GRS below the 25th percentile using a targeted next-generation sequencing library that includes more than 140 genes known to be associated with diabetes. 21% (n=24) were found to have a pathogenic variant in a known diabetes gene. Further tests are ongoing. Our results show that the T1D-GRS is a useful discriminatory tool between diabetes types and a strong predictor of whether a monogenic cause is likely. More importantly, patients with a low T1D-GRS and negative for all known monogenic subtypes should be prioritized for exome or whole genome sequencing to maximize the chance of novel gene discoveries.

Disclosure

M. Sanyoura: None. L.R. Letourneau: Other Relationship; Self; Novo Nordisk Inc.. R.N. Naylor: None. L.H. Philipson: Research Support; Self; Novo Nordisk Inc.. Speaker's Bureau; Self; Merck & Co., Inc.. Research Support; Self; JAEB Center For Health Research, JDRF, Janssen Pharmaceuticals, Inc., Medtronic MiniMed, Inc.. M.N. Weedon: None. R.A. Oram: Other Relationship; Self; Randox. S.W. Greeley: Research Support; Self; Novo Nordisk Inc..
A Nonsynonymous Variant in GLP-1R Is Associated with Decreased Risk of Type 2 Diabetes in Koreans
Authors: KWAK SOO HEON , CHAE JEESOO , YOON JIWON , CHOI SUNGHEE , LIM SOO , MOON MIN KYONG , CHO YOUNG MIN , JANG HAK CHUL , JANG YOON SHIN , KANG HYUN MIN , KIM JONG-IL , PARK KYONG SOO
Published: June 2018

Type 2 diabetes (T2D) is a common complex disorder with strong genetic predisposition. Although different ethnic groups share most of the common variants of T2D, there might be nonsynonymous variants that are specific to an ethnic group. In this study, we used whole exome sequencing (N=917) and exome genotyping array (N=3,026) to identify nonsynonymous variants in Korean T2D cases and controls (stage 1) and validated the findings in an independent set of 15,095 T2D cases and controls (stage 2). Whole exome capture was prepared using Agilent SureSelect version 4+UTR and sequencing was performed by Illumina HiSeq 2000. Nonsynonymous variants were incorporated into customized Axiom Biobank Plus Genotyping array. Genotype imputation was performed with 1,000 Genomes Project phase 3 reference. In silico data look-up was done for stage 2 analyses. EPACTS software was used for association testing and METAL was used for meta-analysis of stage 1 and 2 results. We identified 728,838 variants using whole exome sequencing, which include 258,463 novel variants. A nonsynonymous variant in GLP-1R (rs3765467, R131Q) was associated with decreased risk of T2D in genome-wide significance (OR = 0.85, P = 3.01x10-8). This variant was associated with lower fasting glucose concentration (P = 0.00020) and lower HbA1c level (P = 0.0085) in nondiabetic controls. The minor allele frequency of this variant was 21.1% in Koreans but it was monomorphic in Europeans. We also validated the previous association between PAX4 nonsynonymous variant (rs2233580, R192H) and increased risk of T2D (OR = 1.52, P = 6.41x10-15). This variant was associated with lower age at diagnosis and decreased C-peptide level in T2D cases (P < 0.05). Gene-wise analysis revealed that SLC30A8 was most significantly associated with decreased risk of T2D (P = 0.00010) by Madsen and Browning method.

In summary, we have identified East Asian specific nonsynonymous variants in GLP-1R and PAX4 to be associated with T2D in genome-wide significance.

Disclosure

S. Kwak: None. J. Chae: None. J. Yoon: None. S. Choi: None. S. Lim: None. M. Moon: None. Y. Cho: None. H. Jang: None. Y. Cho: None. H. Kang: None. J. Kim: None. K. Park: None.
The Induction of TRIB3 Causing Glucose Toxicity in Muscle Requires the O-GlcNAcylation of SP1
Authors: LIU XIAOBING , KANG MINSUNG , TIMOTHY GARVEY W.
Published: June 2018

Background: Glucose-induced insulin resistance is mediated by induction of the pseudokinase TRIB3, which requires glucose metabolism via the hexosamine biosynthetic pathway (HBP). We have also shown that the SP1 binding site in the TRIB3 promoter region is essential for glucose’s ability to induce TRIB3 in skeletal muscle. However, the mechanism by which glucose regulates TRIB3 gene expression remains unknown.

Hypothesis: Since the HBP provides the substrate for O-GlcNAcylation, we hypothesize that glucose results in O-GlcNAcylation of SP1 and this modulates induction of TRIB3 causing insulin resistance.

Results: When treated with 5mM or 25mM glucose, the induction of TRIB3 gene expression (P<0.01) in L6 myotubes is accompanied by upregulation of SP1 expression (P<0.01). When fully differentiated L6 myotubes are transiently transfected with SP1 siRNA, knockdown of SP1 produced a corresponding suppression of TRIB3 both in the high and low glucose (P<0.05). To identify whether O-GlcNAcylation is involved, we cultured L6 myotubes in either 5mM or 25mM glucose medium with or without OSMI-1, an inhibitor of rate-limiting O-GlcNAc transferase (OGT). OSMI-1 did not affect glucose’s ability to augment SP1 2-fold (p<0.01), but effectively blocked induction of TRIB3. At the same time, OSMI-1 downregulated O-GlcNAcylation of SP1 and other O-GlcNAc modified proteins.

Conclusion: (1) Increased TRIB3 mRNA expression parallels with upregulated SP1 expression; (2) Glucose induction of TRIB3 mRNA and protein is eliminated when HBP flux and O-GlcNAcylation is disrupted by the inhibition of OGT; (3) SP1 sites are critical in the regulatory action of glucose on the TRIB3 promoter and can be modulated by SP1 O-GlcNAcylation. While further studies are underway, these data provide new insights regarding mechanisms causing glucose-induced insulin resistance, and TRIB3 and TRIB3 promoter regulation are attractive targets for prevention of glucose toxicity in diabetes.

Disclosure

X. Liu: None. M. Kang: None. W. Garvey: Advisory Panel; Self; Novo Nordisk Inc., Merck & Co., Inc.. Research Support; Self; Sanofi, Pfizer Inc., Novo Nordisk Inc., AstraZeneca, Merck & Co., Inc., Elcelyx Therapeutics, Inc., Lexicon Pharmaceuticals, Inc., Eisai Inc..
Serum Anti-Phospholipidase A2 Receptor Antibody Differentiating Membranous Nephropathy from Diabetes Kidney Disease in a Large Population of Type 2 Diabetes Mellitus Patients
Authors: WANG QIAN , DONG ZHEYI , CHEN XIANGMEI
Published: June 2018

Background: With the increase of diabetes, diabetes kidney disease (DKD) was increasing year by year. The pathological types of DKD were various, among which, membranous nephropathy (MN) accounts for a large proportion of nondiabetic renal diseases (NDRD). Recently, serum anti-phospholipidase A2 receptor antibody(sPLA2R-Ab) was used for the diagnosis of MN, the diagnostic efficacy of sPLA2R-Ab for MN in DKD population need to be further verified.

Methods: Screened on patients with type 2 diabetes mellitus (T2DM) who underwent a renal biopsy and have a valid sPLA2R-Ab test results, hospitalized in the general hospital of the people’s liberation army of China between May 1st, 2016 and January 30th, 2018. Eligible patients were divided into two groups according to the pathological results: MN group and non-MN group. Patients’ clinical and laboratory datas were collected. Statistical analysis was performed with SPSS20.0, and P<0.05 was considered statistically significant.

Results: The study included 252 patients: 59 (23.4%) in the MN group and 193 (76.6%) in the non-MN group. MN accounts for 38% of NDRD. The sPLA2R-Ab test results of 35 patients were positive, all of them belonged to MN group. The positive rate of sPLA2R-Ab was 59.3% in MN group and 0 in non-MN group. Patients in the MN group had a higher fasting plasma glucose, age and lower estimated glomerular filtration rate, creatinine and blood pressure as well as a shorter duration of DM than patients in the non-MN group.sPLA2R-Ab achieved a good diagnostic efficiency with a sensitivity of 59.3%, specificity of 100%, negative predictive value of 88.9% and accuracy of 90.5% for MN in DKD.

Conclusion: If the sPLA2R-Ab result of DKD patients is positive, the patient must be MN, and if the result is negative, further renal biopsy should be done. The high accuracy of sPLA2R-Ab may be substituted for renal biopsy partially in the diagnosis of MN in DKD population.

Disclosure

Q. Wang: None. Z. Dong: None. X. Chen: None.
NFIA Colocalizes with PPARgamma and Activates the Cell-Type–Specific Enhancers to Control the Brown Fat Gene Program
Authors: HIRAIKE YUTA , WAKI HIRONORI , TSUTSUMI SHUICHI , ABURATANI HIROYUKI , YAMAUCHI TOSHIMASA , KADOWAKI TAKASHI
Published: June 2018

Brown fat dissipates energy as heat by means of the uncoupling protein-1 (UCP1) on the mitochondrial inner membrane. Brown fat activity is inversely correlated with body mass index in humans, and pilot studies have shown that interventions such as chronic cold exposure successfully recruit human brown fat and increase systemic energy expenditure. Therefore, stimulating development and/or function of brown fat would be a novel strategy for the treatment of obesity and its complications. Here, we identified nuclear factor I-A (NFIA) as a transcriptional regulator of brown fat by a genome-wide open chromatin analysis of murine brown and white fat followed by motif analysis of the brown-fat-specific open chromatin regions. Introduction of NFIA into myoblasts results in brown adipocyte differentiation. Conversely, the brown fat of NFIA knockout mice displays impaired expression of the brown-fat-specific genes and reciprocal elevation of muscle genes. In human brown fat, expression of NFIA and the brown-fat-specific genes including UCP1 is positively correlated. Mechanistically, NFIA co-localizes with the master transcriptional regulator of adipogenesis, PPARgamma, selectively at the brown-fat-specific enhancers. Moreover, the binding of NFIA precedes and facilitates the binding of PPARgamma, leading to increased chromatin accessibility and active transcription. Most recently, we performed mass spectrometry analysis of NFIA protein complex to identify and characterize the crucial partner of NFIA-dependent brown-fat-specific chromatin regulation. Collectively, these results indicate that NFIA is a key transcription factor that activates the cell-type-specific enhancers and facilitates the binding of PPARgamma for controlling the brown fat gene program.

Disclosure

Y. Hiraike: Speaker's Bureau; Self; Novo Nordisk A/S. Research Support; Self; Novo Nordisk A/S, Mitsubishi Tanabe Pharma Corporation. H. Waki: Research Support; Self; MSD K.K., Novartis Pharma K.K., Boehringer Ingelheim Pharmaceuticals, Inc., Sanofi K.K., Ono Pharmaceutical Co., Ltd., Kyowa Hakko Kirin Co., Ltd., Takeda PHarmaceutical Co., Ltd.. S. Tsutsumi: None. H. Aburatani: None. T. Yamauchi: Research Support; Self; Novo Nordisk Inc., Daiichi Sankyo Company, Limited, Sanofi, Takeda, Tanabe-Mitsubishi. T. Kadowaki: Consultant; Self; Novo Nordisk A/S, AstraZeneca, Merck Sharp & Dohme Corp.. Research Support; Self; Kissei Pharmaceutical Co., Ltd., Taisho Pharmaceutical Co., Ltd., Sanofi, Kyowa Hakko Kirin Co., Ltd., Novo Nordisk A/S, Astellas Pharma, Daiichi Sankyo Company, Limited, Takeda, Mitsubishi Tanabe Pharma Corporation, Ono Pharmaceutical Co., Ltd., Merck Sharp & Dohme Corp., Nippon Boehringer Ingelheim Co. Ltd.. Speaker's Bureau; Self; Astellas Pharma, AstraZeneca, Merck Sharp & Dohme Corp., Ono Pharmaceutical Co., Ltd., Takeda, Eli Lilly and Company, Nippon Boehringer Ingelheim Co. Ltd., Novo Nordisk A/S.
Fasting-Induced Changes in Glucagon Secretion Are Dysregulated in Obesity
Authors: STERN JENNIFER H. , SMITH GORDON I. , UNGER ROGER H. , KLEIN SAMUEL , SCHERER PHILIPP E.
Published: June 2018

Hyperglucagonemia is a hallmark in obesity and type 2 diabetes (T2DM). Suppression of glucagon signaling improves glycemic control in T2DM. We evaluated glucagon homeostasis in lean and obese mice and people. Discordant with the canonical rise in glucagon with fasting, our studies show that fasting (4, 8, 16 and 24 h) caused a progressive decrease in serum glucagon in diet-induced obese, hyperglucagonemic mice (P<0.01), yet a progressive increase in serum glucagon in lean mice (P<0.01). Serum insulin decreased with fasting in both lean and obese mice (P<0.01). Accordingly, fasting increased the glucagon:insulin ratio in the lean mouse (P<0.01), but did not affect the glucagon:insulin ratio in the obese mouse. Two hours of re-feeding restored hyperglucagonemia in obese mice (P<0.01). Pancreatic perfusion studies in obese, hyperglucagonemic mice confirm that 16 h of fasting decreases pancreatic glucagon secretion (P<0.01). Consistent with our findings in the mouse, fasting decreased (P<0.05) serum glucagon in obese participants. In contrast, fasting increased serum glucagon concentrations in lean participants (P<0.05). As expected, fasting decreased serum insulin in both lean and obese participants (P<0.05 for both). As a result, fasting induced a more robust rise in the glucagon:insulin ratio in lean compared to obese participants (P<0.01). In addition, mixed meal feeding increased serum glucagon in people with obesity. These findings suggest that the metabolic pathophysiology of obesity may be driven by inappropriate meal-induced regulation of glucagon, resulting in a relatively static glucagon:insulin ratio.

Disclosure

J.H. Stern: None. G.I. Smith: None. R.H. Unger: None. S. Klein: Stock/Shareholder; Self; Aspire Bariatrics. Consultant; Self; Pfizer Inc.. Research Support; Self; Merck & Co., Inc., Johnson & Johnson Services, Inc., REMD Biotherapeutics. P.E. Scherer: None.
CRISPR-Engineered Human Brown-Like Adipocytes Promotes Metabolic Homeostasis through Activation of Endogenous Brown Fat
Authors: WANG CHIH-HAO , LUNDH MORTEN , KRISZT ROKUS , HUANG TIAN LIAN , LEIRIA LUIZ O. , SHAMSI FARNAZ , LYNES MATT , DARCY JUSTIN , EMANUELLI BRICE , KIEBISH MICHAEL , TSENG YU-HUA
Published: June 2018

Brown and brown-like beige adipocytes dissipate energy for thermogenesis and have been proposed to combat obesity and metabolic disorders. Uncoupling protein 1 (UCP1) is uniquely expressed in brown and beige adipocytes, and facilitates fuel utilization and energy expenditure. Ectopic UCP1 overexpression improves obesity and insulin resistance in mice. Recently, we utilized the CRISPR/Cas9 synergistic activation mediator and specific gRNAs to boost endogenous UCP1 expression in human white adipocytes. The CRISPR/Cas9 engineered human brown fat-like (referred to as HUMBLE, hereafter) cells had acquired human brown fat features when comparing bona fide human brown adipocytes derived from the same individual. Obese mice receiving a transplantation of HUMBLE cells showed improved glucose tolerance and insulin sensitivity, as well as increased energy expenditure. This was mediated, at least in part, via activation of endogenous brown fat, which showed an increase in heat production and glucose uptake in mice receiving HUMBLE cells. Co-culture with HUMBLE cells or treatment of conditional medium from HUMBLE cells promoted glucose uptake and thermogenesis in brown adipocytes. Metabolomic analysis identified an increased ratio of arginine and asymmetrical dimethylarginine (ADMA) in the sera of mice receiving HUMBLE cell transplantation. Arginine is known to promote nitric oxide biosynthesis, which can activate thermogenic function of brown fat. Consistently, the level of nitric oxide was increased in the endogenous brown fat of mice receiving HUMBLE cells. Inhibition of nitric oxide synthase abolished the HUMBLE cells-mediated activation of brown adipocytes in vitro and in vivo.

In summary, CRISPR/Cas9-mediated activation of endogenous UCP1 expression in human white adipocytes creates brown fat-like features, and activates endogenous brown fat, potentially through arginine-related metabolites to improve the systemic glucose utilization.

Disclosure

C. Wang: None. M. Lundh: None. R. Kriszt: Employee; Self; QuintilesIMS. T. Huang: None. L.O. Leiria: None. F. Shamsi: None. M. Lynes: None. J. Darcy: None. B. Emanuelli: None. M. Kiebish: None. Y. Tseng: Other Relationship; Self; Chugai Pharmaceutical Co., Ltd.. Research Support; Self; MedImmune.
Targeting Intestinal Macrophages as a Potential Therapeutic Option in Obesity
Authors: ROHM THERESA V. , ALASFOOR SHEFAA , BOSCH ANGELA J. , CAVELTI-WEDER CLAUDIA
Published: June 2018

Background: Chronic inflammation such as systemic or tissue inflammation has been well established in metabolic disease. Although macrophages thereby play a key role, not much is known about the initiation of inflammation. As the gut makes up the largest macrophage reservoir of the body and gastrointestinal changes occur in metabolic disease (altered gut microbiota, increased endotoxin and cytokines), the aim of our study was to assess the role of intestinal macrophages (iMϕs) in obesity.

Research Design and Method: IMϕs were isolated from the colon of C57BL/6, germ-free or CCR2-/- mice fed a high fat diet (HFD) or control diet up to three months and characterized by flow cytometry as CCR2+ (“inflammatory”) or CCR2- (resident, anti-inflammatory) subpopulations. For dose-dependent iMϕ depletion, HFD-fed mice were orally treated with 50, 100 or 200 µg/g CSF-1R inhibitor (BLZ945) or its vehicle for up to ten weeks.

Results: We found that within one week and up to three months of HFD, inflammatory iMϕs increase, preceding adipose tissue inflammation. Two mouse models protected from metabolic disease - germ-free and CCR2-/- mice - exhibited ≥10-fold lower inflammatory iMϕ numbers when compared to WT mice, suggesting that a threshold of iMϕs is required to elicit metabolic disease. Based on this finding, we depleted macrophages by increasing doses of CSF-1R inhibitor and observed a gradual iMϕ-reduction, which correlated with a dose-dependent improvement in fasting glucose, glucose tolerance and insulin levels.

Conclusion: HFD-induced obesity is associated with an increase in inflammatory iMϕs, while mouse models protected from metabolic disease have lower numbers. Accordingly, dose-dependent depletion of iMϕs is accompanied by gradual improvements in glucose metabolism. This suggests that the number of iMϕs is involved in initiation of metabolic disease and thus could serve as a potential therapeutic target in obesity.

Disclosure

T.V. Rohm: None. S. AlAsfoor: None. A.J. Bosch: None. C. Cavelti-Weder: None.
Liraglutide Suppresses Atherosclerosis via AMP-Activated Protein Kinase Dependent and Independent Mechanisms in Diabetic Apolipoprotein Enull Mice
Authors: KOSHIBU MASAKAZU , MORI YUSAKU , KUSHIMA HIDEKI , HIROMURA MUNENORI , KOHASHI KYOKO , TERASAKI MICHISHIGE , HIRANO TSUTOMU
Published: June 2018

Background: Glucagon like peptide 1 receptor agonists (GLP-1RAs) have anti-atherosclerotic properties, possibly through activation of AMP-activated protein kinase (AMPK) in the vasculature. However, their relationship remains largely unknown.

Methods: Streptozotocin-induced diabetic apolipoproteinE-null mice (male, 20 w) were randomly assigned to six treatment groups: saline (control, C), liraglutide at low or high doses (L- and H-Lira at 17 and 107 nmol/kg/d, respectively), AMPK inhibitor dorsomorphin (AMPKI, 25 mg/kg/d), AMPKI+L-Lira, or AMPKI+H-Lira.

Results: First, we confirmed that AMPKI inhibited H-Lira-induced AMPK phosphorylation in the aorta (1.5-fold). After 4 w, biological parameters were similar, except for HbA1c levels (C 9.8, L-Lira 8.6, H-Lira 8.4%, p<0.05). Both L- and H-Lira suppressed intraplaque macrophage accumulation at the aortic sinus by 60%. H-Lira also suppressed aortic surface plaque area by 40%, which was not significant in L-Lira (25%). In AMPKI-treated mice, biological parameters were similar. AMPKI completely eliminated L-Lira-induced anti-atherogenic effects but did not attenuate H-Lira-induced suppression of atherosclerotic area (38%) and intraplaque macrophage accumulation (63%), suggesting that AMPK mechanisms are L-Lira-dependent and H-Lira-independent. In cultured human vascular endothelial cells, Lira (100 nM) reduced TNF-induced inflammatory cytokine and adhesion molecule expression by 60 to 80%, which was retained at more than 50% after siRNA-knockdown of AMPK (p<0.05). In cultured human monocytic cells (U937), Lira suppressed LPS-induced inflammatory cytokine expression by 30% (p<0.05), which was completely reversed by AMPKI.

Conclusions: Lira suppressed atherosclerosis via AMPK-dependent and -independent mechanisms; AMPK dependency is altered by GLP-1RA dose and cell type.

Disclosure

M. Koshibu: None. Y. Mori: None. H. Kushima: None. M. Hiromura: None. K. Kohashi: None. M. Terasaki: None. T. Hirano: Speaker's Bureau; Self; Novo Nordisk Inc., AstraZeneca.
Differential Effects of the Insulin-Only and Bihormonal Configurations of the Bionic Pancreas on Mean Glucose and Hypoglycemia during the Daytime and Nighttime
Authors: SHERWOOD JORDAN , BALLIRO COURTNEY A. , JAFRI RABAB Z. , EL-KHATIB FIRAS , MAHENO MICHELE , HILLARD MALLORY A. , O'DONOVAN ALEXANDER J. , SELAGAMSETTY RAJENDRANATH , ZHENG HUI , DAMIANO EDWARD , RUSSELL STEVEN J.
Published: June 2018

The bihormonal bionic pancreas (BP) reduced both mean CGM glucose (CGMG) and time in the hypoglycemic range relative to usual care in outpatient studies. We recently performed an outpatient study in which we compared insulin-only (IOBP) and bihormonal (BHBP) configurations of the BP with usual care (UC; conventional or sensor-augmented insulin pump therapy) both with and without remote monitoring. We previously reported that monitoring did not affect the mean CGMG or hypoglycemia with either configuration of the BP. The current analysis focuses on the data collected from arms without remote monitoring (how the BP is eventually meant to be used) to determine how the effects of the BP differ between the day and night. As shown in the Table, both the IOBP (with glucose target of 110 mg/dl) and BHBP (with glucose target of 100 mg/dl) significantly reduced mean CGMG vs. UC during both the daytime and nighttime. In contrast, only the BHBP significantly reduced the percent of time spent with CGMG <60 mg/dl vs. UC. The BHBP similarly reduced percent of time <60 mg/dl vs. IOBP. The significant reductions in hypogycemia associated with the BHBP were driven primarily by reductions in hypoglycemia at night.

Disclosure

J. Sherwood: None. C.A. Balliro: None. R.Z. Jafri: None. F. El-Khatib: Stock/Shareholder; Self; Beta Bionics. Employee; Self; Beta Bionics. M. Maheno: None. M.A. Hillard: None. A.J. O'Donovan: None. R. Selagamsetty: None. H. Zheng: None. E. Damiano: Other Relationship; Self; Beta Bionics. S.J. Russell: Other Relationship; Self; Beta Bionics, Novo Nordisk Inc.. Advisory Panel; Self; Companion Medical, Tandem Diabetes Care, Inc., Unomedical a/s. Research Support; Self; Beta Bionics, Zealand Pharma A/S, MITRE Corporation.
NOX, NOX, Are You Here? The Emerging Role of NOX5 in Diabetic Neuropathy
Authors: EID STEPHANIE , HAYES JOHN M. , GUO KAI , PACUT CRYSTAL , MENDELSON FAYE , HUR JUNGUK , FELDMAN EVA L.
Published: June 2018

Diabetic neuropathy (DN) is one of the most common complications of diabetes worldwide, affecting up to 60% of all diabetic patients. Although the pathogenesis of DN is poorly understood, it is widely accepted that reactive oxygen species (ROS) mediate in part the cellular and molecular injury observed in DN. NADPH oxidase (Nox) enzymes generate ROS and of the 7 isoforms, NOX5 is present only in man. In this study, we aimed to investigate a role for NOX5 in DN by identifying the cellular localization, methylation status and expression levels of NOX5 in cutaneous nerve fibers and sural nerve biopsies of diabetic patients. Cellular localization of NOX5, myelin basic protein (MBP) and protein gene product (PGP) 9.5 were determined in cutaneous nerve fibers of controls and subjects with DN. NOX5 methylation status and protein levels were assessed in subjects with DN that were divided into two groups based on changes in sural nerve myelinated fiber density: regenerators (showing significant nerve regeneration) and degenerators (showing significant nerve degeneration). NOX5 expression was present in deep myelinated cutaneous fibers. Bisulfite sequencing revealed that the NOX5 promoter, enriched with CpG sites, is hypomethylated in sural nerve biopsies of the degenerator cohort compared to the regenerator. The increase in NOX5 expression in degenerator sural nerves was accompanied with a decrease in MBP levels relative to the regenerator group. Our results highlight that in subjects with DN with sural nerve degeneration, there is hypomethylation of the NOX5 promoter associated with an increase in its protein expression, suggesting a potential epigenetic and mechanistic role of NOX5 in DN.

Disclosure

S. Eid: None. J.M. Hayes: None. K. Guo: None. C. Pacut: None. F. Mendelson: None. J. Hur: None. E.L. Feldman: None.
IMG-1 Activates and Mobilizes Insulin-Producing Islet Progenitor Cells (CD133+/Insulin+) In Vitro
Authors: BOTTINO RITA , POLLETT JONATHAN B. , THAI NGOC , TRUCCO MASSIMO
Published: June 2018

IMG-1 is a novel therapeutic for the treatment of types 1 and 2 diabetes that may have islet regenerative properties. 300 islets isolated from mouse pancreases were cultured in standard media with IMG-1 (0.5µg/mL) and compared with control non-treatment. Within 24 hours, the IMG-1 treated islets became smaller (less clustered) and islet cells appeared to actively migrate away from the clusters. Control islets maintained their clustered morphology and did not show any evidence of cell migration. Three days after IMG-1 treatment, there were both free-floating and attached cells in the culture. The free-floating cells were 30% viable and the attached cells and leftover islets were 85% viable. In contrast, control islet cultures had neither free-floating nor attached cells although islet clusters looked healthy (cell viability >90%). By day 10 of IMG-1 treatment, islet clusters were few in numbers (<10%), no free-floating cells remained, and the attached cells appeared to form colonies. Total cell viability was >90%. Control cells still had no migration or attachment, although within the islet clusters cell viability remained around >90%. At 25 days, islet cells from both cultures were fixed and stained for the progenitor cell marker, CD133, and insulin. IMG-1 treated colonies were 75% insulin-positive by immunohistostaining. Furthermore, these cells were 70% CD133-positive with 60% of the culture being CD133/Insulin double-positive. In contrast, non-treated islets were 90% insulin+ cells, but <5% CD133+ and <5% double positive (CD133/insulin). IMG-1 has a novel effect on islets and induces the activation and migration of islet cells out of the islet. These cells form colonies of CD133/insulin double-positive cells that putatively represent activated beta-cell progenitor cells that can be used to regenerate damaged islets.

Disclosure

R. Bottino: None. J.B. Pollett: Employee; Self; Imagine Pharma. N. Thai: Board Member; Self; Imagine Pharma. M. Trucco: None.
Thymic Development of Beta-Cell Specific Regulatory T Cells in Neonatal NOD Mice
Authors: LIN YONG , LEE THOMAS , BETTINI MARIA , BETTINI MATTHEW
Published: June 2018

Type 1 diabetes (T1D) is a T cell-mediated autoimmune disease that involves the destruction of insulin producing β cells found within the pancreatic islets of Langerhans. Recent attention to neonatal thymocyte development has revealed a window during which central tolerance is established in an Autoimmune Regulator (Aire) dependent manner. However, expression of Chromogranin A (ChgA) within the thymus has not been detected; therefore, tolerance to ChgA may be due in part to peripheral dendritic cells (DCs) presenting ChgA to developing thymocytes. The relative contribution of thymically and peripherally derived antigens in the generation of neonatal β cell antigen specific CD4+ regulatory T cells (Tregs) is not well understood. For example, post-translational modification (PTM) of ChgA by fusion with an insulin peptide increases epitope immunogenicity by forming neo-antigens. Pathogenic T cells that are specific for neo-antigens uniquely expressed in the pancreas may escape thymic selection. Therefore, we utilized two experimental designs to test the role of peripheral antigen exposure on neonatal development of β cell specific Tregs. Firstly, we adoptively transferred dendritic cells pulsed with ChgA peptides into BDC2.5 TCR Tg mice and WT NOD mice to study the role of migratory DCs on neonatal development of ChgA Tregs. Secondly, we targeted thymic Langerin+ dendritic cells with anti-Langerin linked to ChgA peptides. In both experimental approaches, we found a robust increase in the ratio and number of ChgA specific Tregs in the presence of the hybrid insulin/chromogranin A peptide (2.5HIP) while also observing an increase in negative selection. The expansion of thymic Tregs was observed in both adult and neonatal mice. Together our data suggests neonatal exposure to PTM peptides enhances thymic development of β cell specific Tregs and may alter T1D pathogenesis.

Disclosure

Y. Lin: None. T. Lee: None. M. Bettini: None. M. Bettini: None.