bluuebunny/crossref_metadata_2025_split huggingface dataset:
303-OR: ADA Presidents' Select Abstract: Transethnic Association Study of Type 2 Diabetes in More than a Million Individuals
Authors: MAHAJAN ANUBHA , SIM XUELING , ZHANG WEIHUA , BELOW JENNIFER E. , KITAJIMA HIDETOSHI , SPEIDEL LEO , MORRIS ANDREW 
Published: June 2019

We conducted the largest trans-ethnic meta-analysis of genome-wide association studies of type 2 diabetes (T2D) by aggregating 171,262 cases and 1,075,072 controls from diverse populations (doubling the effective sample size from recently published European-specific analysis). We identified 250 loci associated with T2D at genome-wide significance (p&lt;5x10-8; 59 novel), which were delineated to 475 distinct signals (p&lt;10-5). Allelic effects on T2D at index variants for most signals were homogenous across populations: only 17% displayed evidence of effect-size heterogeneity related to ancestry (pHET &lt;1.4x10-4, Bonferroni correction). T2D-associated variants were jointly enriched (p&lt;0.00023, correction for 220 annotations) in coding exons, pancreatic islet enhancers and promoters, adipose enhancers, liver enhancers, and binding sites for transcription factors (NKX2.2, PDX1, FOXA2, EZH2). Compared to previous efforts, increased sample size, population diversity, and annotation-informed fine-mapping appreciably improved localisation of causal variants achieving single variant resolution at 217 signals (&gt;50% of the posterior probability of association assigned to a single variant). Through co-localisation of these high-confidence causal variants with protein and RNA QTLs (in disease-relevant tissues), and chromatin conformation capture data (in human islets), we identified &gt;80 candidate effector genes including several not previously implicated e.g., CARD9, PDE8B, ABCB10, SIX3, INHBB. Genetic risk scores derived from ethnic-specific effect sizes at lead variants were transferrable across populations, with minimum liability-scale variance explained from Europeans into Africans. T2D-associated variants displayed evidence of directional adaptation through time, indicating selection for increased T2D risk in African populations, relative to elsewhere in the world. These analyses represent the most comprehensive view of the genetic contribution to T2D to date.

Disclosure
A. Mahajan: None. X. Sim: None. W. Zhang: None. J.E. Below: None. H. Kitajima: None. L. Speidel: None. A. Morris: None.
304-LB: Connection between Dynamics of Lipid Droplets and Energy Expenditure in Adipose Tissue
Authors: YANG LI , LI XIN , SUN KAI 
Published: June 2019

Lipid droplets (LDs), which largely found in adipocytes, are major organelles for energy storage and burndown. It is well known that ß-adrenergic signaling pathway liberates fatty acid from LDs to activate and fuel the thermogenesis program in mitochondria. However, the detailed events on the surface of LDs remain unclear. Here, we identified a novel hydrolase named Carboxylesterase 3 (Ces3) that targets LDs upon cold exposure by LC-MS/MS. We further found that translocation of Ces3 to LDs is cAMP-PKA pathway dependent. To investigate the function of Ces3 in β-adrenergic-activated adipocytes, we suppressed Ces3 with WWL229 (a specific Ces3 inhibitor) and small interfering RNA (siRNA) in 3T3-L1 and BAC (immortalized brown adipocytes) cells. We found that blockage of Ces3 by both methods dramatically attenuates ISO-induced lipolysis in cells. Furthermore, Ces3 inhibition leads to impaired mitochondrial respiratory function and attenuates ISO-induced thermogenic program in adipocytes by down-regulating Ucp1 and Pgc1α genes via PPARγ. Importantly, we further confirmed our findings by in vivo assays in mice. Specifically, our data showed that the thermogenic program in adipose tissue upon cold exposure is downregulated in WWL229 treated wild type mice and adipose tissue-specific Ces3 knockout mice. As the result, the mice exhibit dramatically impaired cold tolerance. Furthermore, we found that serum collected from cold-exposed wild type mice massively increase UCP1 levels in BAC cells while serum from Ces3 deficient mice fails to do so.
In conclusion, our study highlighted the Ces3-mediated lipolytic signaling as a unique process to regulate thermogenesis in adipose tissue.

Disclosure
L. Yang: None. X. Li: None. K. Sun: None.

Funding
National Institutes of Health
307-LB: IL1R1 Mediates Celastrol’s Leptin-Sensitization and Antiobesity Effects
Authors: FENG XUDONG 
Published: June 2019

Celastrol, isolated from the root extracts of thunder god vine, is the most potent anti-obesity agent that has been reported to date. Celastrol increases leptin sensitivity in the hyperleptinemic diet-induced obese (DIO) mice and leads to 45-50% weight loss in this model, with either no or only marginal effects in db/db, ob/ob or lean mice. In this work, we first showed that lipocalin 2 (LCN2) is the most upregulated gene in the hypothalamus of DIO mice after celastrol treatment. Celastrol treatment leads to a significant upregulation of LCN2 both in bone marrow, plasma, and the hypothalamus. However, LCN2 does not play a significant role in suppressing appetite and reducing body weight or mediating celastrol’s antiobesity effects. Secondly, by using systems biology approaches for analysis of temporal regulations of the hypothalamic transcriptome of DIO, lean and db/db mice that were treated with celastrol, we have identified interleukin 1 receptor 1 (IL1R1) as a mediator of celastrol action and documented that IL1R1-deficient mice is completely resistant to celastrol’s leptin sensitization, anti-obesity, antidiabetic and anti-NASH effects. Thus, IL1R1 is a gate-keeper for celastrol’s metabolic actions.

Disclosure
X. Feng: None.

Funding
Fidelity Biosciences Research Initiative
308-LB: A Short Period of Exposure to Air Pollution (PM2.5) Is Sufficient to Induce Dysbiosis, Hyperphagia, and Fat Mass Gain
Authors: ZORDÃO OLIVIA P. , SANTOS ANDREY , CAMPOLIM CLARA MACHADO , LIMA RAQUEL ATAÍDE , FERREIRA CLÍLTON KRAüSS , SAAD MARIO , SALDIVA PAULO , VERAS MARIANA , PRADA PATRICIA O. 
Published: June 2019

Fine particulate matter (PM2.5) is the most harmful component of air pollution and is associated with an increased risk of cardiometabolic diseases, such as T2DM. The small diameter of PM2.5 allows it to be inhaled and subsequently invades tissues through the airways. Intake of PM2.5 is also possible even if this is not the primary route of entry of PM2.5 into the body. However, it is still unclear whether ingestion and inhalation of PM2.5 in a short time would be sufficient to alter the gut microbiota and energy metabolism. Thus, we aimed to investigate whether 1 day or 5 days of exposure to PM2.5 would be enough to alter energy metabolism and the composition of gut microbiota at the phylum level. Male C57BL/6J mice on a chow diet were exposed to PM2.5 in a fixed dose (600 μg/m3/day) or filtered air (FA) for 1 day or 5 days using the Harvard Ambient Fine Particle Concentrator. The 1-day exposure to PM2.5 did not change body weight, food intake or energy expenditure, but altered the composition of the gut microbiota compared to the FA group. In the gut, there was a reduction of Bacteroidetes and Verrucomicrobia and an increase of Tenericutes. After 5 days of exposure to PM2.5, there was an increase in food intake, fat mass. In the gut, there was an increase in Firmicutes and in the ratio of Firmicutes/Bacteroidetes after 5 days of exposure to PM2.5 compared to the FA group. Thus, after a short period of exposure to air pollution, there was a significant change in the gut microbiota, beginning with the reduction of the Bacteroidetes and Verrucomicrobia phyla and the increase of the phylum Tenericutes with 1 day of exposure. This result occurred without altering the energy balance. A robust dysbiosis installation happened after 5 days of exposure to PM2.5, Firmicutes/Bacteroidetes ratio was elevated and associated with fat mass gain, and hyperphagia.

Disclosure
O.P. Zordão: None. A. Santos: None. C.M. Campolim: None. R. Ataíde Lima: None. C.K. Ferreira: None. M. Saad: None. P. Saldiva: None. M. Veras: None. P.O. Prada: None.

Funding
Sao Paulo Research Foundation (2017/18498-62017/19703-22017/11518-1)
310-OR: Integrating Islet and Skeletal Muscle Enhancer RNA Signatures with Diverse Omics Data Enables Fine-Mapping of Diabetes and Related Trait GWAS Loci
Authors: PARKER STEPHEN , VARSHNEY ARUSHI ,   FUSION TISSUE STUDY
Published: June 2019

Type 2 diabetes (T2D) results from interaction between genetic variation and environmental exposures that influence insulin secretion in pancreatic islets and insulin resistance in skeletal muscle and other peripheral tissues. Genome-wide association studies (GWAS) have identified hundreds of loci that modulate risk for T2D and related insulin traits. These loci mostly occur in non protein-coding regions which makes identification of causal regulatory single nucleotide polymorphisms (SNPs) and their associated mechanisms difficult. Previous studies show that many active enhancers are transcribed into enhancer RNA (eRNA). To identify actively transcribed regulatory elements with high genomic resolution and prioritize SNPs, we generated eRNA profiles using cap analysis of gene expression (CAGE) across 57 islet and 146 skeletal muscle samples and 15 replicates of the human beta cell line EndoC-BH1. We computationally integrated these CAGE profiles with ChIP-seq profiles for five histone marks along with open chromatin ATAC-seq profiles to generate high-resolution cross-tissue chromatin states. We identify multiple CAGE-enriched chromatin states that correspond to actively transcribed promoter or enhancer regions, which are supported by diverse patterns of chromatin activity across an orthogonal set of 127 human epigenome maps. We find 11,856 CAGE+ promoter regions across islets and muscle and 54% are shared across the two tissues, whereas we observe 1,712 CAGE+ eRNA regions and only 7% are shared, indicating the cell-identity nature of these eRNA states. We find significant enrichment for GWAS and eQTL SNPs to overlap these CAGE+ chromatin states. Finally, at a subset of loci, we validate these findings using a modified high-throughput STARR-seq assay where we observe allelic activity. These results provide the highest-resolution epigenome maps for islets and muscle to date, and we are using them to fine-map GWAS SNPs.

Disclosure
S. Parker: None. A. Varshney: None.

Funding
American Diabetes Association/Pathway to Stop Diabetes (1-14-INI-07 to S.P.); National Institutes of Health (1R01DK117960)
317-OR: Spatio-temporal Signature of Hypothalamic Transcriptome in Obesity
Authors: TANAKA TOMOHIRO 
Published: June 2019

Background: Hypothalamus is a constellation of nuclei, each of which controls an aspect of “internal milieu,” including satiety and energy homeostasis. Here we address molecular signature of each energy-regulating hypothalamic nucleus and its dynamism in diet-induced obesity (DIO).
Methods: We micro-dissected arcuate (ARC), paraventricular (PVN) and lateral (LH) hypothalamic nuclei from mice and verified by the expression of nucleus-specific genes. High fat diet (60%kcal)-fed and control B6 mice (3days, 2, 6 and 16weeks after the start of diets) and 9-week-old ob/ob mice were used. RNA was extracted for RNA-Seq analysis. GO analysis by David.
Results: ARC-specific genes were associated with GO terms: “neuropeptide,” “cilia,” while PVN with “neurogenesis” and LH with “ion channel,” “cell junction.” Among ∼6000 genes whose expression is altered in one or more of 3 nucleus x 4 timepoint = 12 settings in DIO were clustered into 13 groups with one big group of genes suppressed throughout (from 3d to 16w) and 12 groups of upregulated genes, each corresponding to 3 nucleus x 4 timepoint conditions. Genes in each group were specifically upregulated only in one condition and significantly associated with distinct GO terms, suggesting a nucleus x timepoint-specific pathology during the development of DIO. Among altered genes, we selected those with expression highly associated with leptin resistance in ARC, id est, upregulated at 6weeks and thereafter when leptin resistance is fully developed, and downregulated in ob/ob mice, a leptin-sensitive model. Among these was X that encodes a secreted protein whose expression and induction in DIO was confined to ARC. Recombinant X protein induced leptin resistance both in in vitro and in vivo leptin sensitivity assays.
Conclusion: By a comparative transcriptomic approach, “Stages” of DIO can now be defined by nucleus and timepoint-specific gene clusters with specific functions. We further identified a gene with a potential pathogenetic role in leptin resistance in obesity.

Disclosure
T. Tanaka: Research Support; Self; Japan Society for the Promotion of Science. Speaker's Bureau; Self; AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Daiichi Sankyo Company, Limited, Kissei Pharmaceutical Co., Ltd., Kowa Pharmaceutical Europe Co. Ltd., Mitsubishi Tanabe Pharma Corporation, Novartis Pharmaceuticals Corporation, Taisho Toyama Pharmaceutical Co. Ltd., Takeda Pharmaceutical Company Limited.

Funding
Japan Society for the Promotion of Science, Takeda Pharmaceutical Co. Ltd.
322-OR: Risk Factors for Peripheral and Cardiovascular Autonomic Neuropathy in Type 1 Diabetes: Thirty Years of Follow-Up in DCCT/EDIC
Authors: BRAFFETT BARBARA , GUBITOSI-KLUG ROSE , ALBERS JAMES W. , FELDMAN EVA L. , MARTIN CATHERINE , WHITE NEIL H. , ORCHARD TREVOR J. , LOPES-VIRELLA MARIA F. , RASKIN PHILIP , POP-BUSUI RODICA ,   THE DCCT/EDIC STUDY GROUP
Published: June 2019

The Diabetes Control and Complications Trial/Epidemiology of Diabetes Interventions and Complications (DCCT/EDIC) study reported that intensive glucose control as assessed by hemoglobin A1c (HbA1c) reduced the risk of diabetic peripheral neuropathy (DPN) and cardiovascular autonomic neuropathy (CAN) in type 1 diabetes (T1D), with long lasting benefits. We evaluated additional clinical parameters and risk factors that could contribute to the development of DPN and CAN in this cohort. DPN was assessed at three different times and defined as a composite of symptoms, signs, and abnormal nerve conduction studies in ≥2 nerves; CAN was assessed seven different times and defined based on R-R variation, Valsalva maneuver, and postural changes in blood pressure. Generalized estimating equation models were used to evaluate the association of DPN and CAN with individual risk factors over repeated time points spanning 30 years. Among 1,441 DCCT/EDIC participants, 32% developed DPN and 44% developed CAN. Higher mean HbA1c was the most significant risk factor for DPN (OR=1.56 per 1%, 95% CI 1.41,1.72). DPN was also associated with older age (1.43 per 5 years, 95% CI 1.31,1.55), longer duration of T1D, higher albuminuria, β-blocker use, higher mean diastolic blood pressure, and higher HbA1c at DCCT eligibility. CAN was associated (in order) with older age (1.51 per 5 years, 95% CI 1.40,1.63), longer duration of T1D (1.07 per 1 year, 95% CI 1.05,1.10), any sustained microalbuminuria (AER ≥30 mg/24 hour on two consecutive visits), higher mean HbA1c, higher mean and current pulse rate (likely as an indirect measure of CAN), higher mean systolic blood pressure, β-blocker use, any eGFR &lt;60 mL/min/1.73 m2, higher HbA1c at DCCT eligibility, and current cigarette smoking. Higher mean HbA1c was the strongest risk factor for DPN, but not of CAN. These findings help identify patient phenotypes and modifiable risk factors for personalized approaches to neuropathy prevention.

Disclosure
B. Braffett: None. R. Gubitosi-Klug: None. J.W. Albers: Consultant; Self; Eli Lilly and Company. E.L. Feldman: None. C. Martin: None. N.H. White: None. T.J. Orchard: Consultant; Self; Boehringer Ingelheim International GmbH. M.F. Lopes-Virella: None. P. Raskin: Research Support; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Gan & Lee Pharmaceuticals, Novo Nordisk A/S. R. Pop-Busui: Consultant; Self; Novo Nordisk A/S. Research Support; Self; AstraZeneca. Other Relationship; Self; American Diabetes Association.

Funding
Division of Diabetes Endocrinology & Metabolic Diseases (U01DK094176, U01DK094157); National Eye Institute; National Institute of Neurologic Disorders and Stroke; General Clinical Research Centers Program; Clinical Translational Science Center
325-OR: Early Parallel Progression of Peripheral and Cardiac Autonomic Nerve Dysfunction in Recent-Onset Type 1 Diabetes Patients
Authors: BÖNHOF GIDON J. , STROM ALEXANDER , ZAHARIA OANA P. , SZENDROEDI JULIA , MUESSIG KARSTEN , RODEN MICHAEL , ZIEGLER DAN 
Published: June 2019

We previously demonstrated an early parallel involvement of small and large fibers in recent-onset type 2 diabetes. Here we hypothesized that this pattern may also be pertinent to type 1 diabetes (T1D). Therefore, motor and sensory nerve conduction velocity (MNCV, SNCV), vibration perception thresholds (VPT), thermal detection thresholds (TDT), intraepidermal nerve fiber density (IENFD), and heart rate variability (HRV) were assessed in participants with T1D from the German Diabetes Study (GDS) at baseline (diabetes duration ≤1 year) and in glucose-tolerant controls: CON/T1D-B: n=96/360; age [median (1st; 3rd quartile)]: 34.5 (26.0; 46.8)/34.6 (26.5; 45.3) years; male: 72/58%; BMI: 25.0 (22.9; 28.3)/24.0 (22.0 (22.0; 27.0) kg/m²; diabetes duration: -/173 (114; 173) days; HbA1c: 5.1 (5.0; 5.3)/6.4 (5.9; 7.2)%; M-value (hyperinsulinemic-euglycemic clamp): 11.6 (9.1; 13.6)/8.2 (6.5; 10.4) mg*kg-1*min-1. T1D-B showed lower peroneal MNCV ([mean±SEM]: 45.8±0.2 vs. 47.0±0.3 m/s), median MNCV (55.1±0.2 vs. 56.0±0.7 m/s), and IENFD (10.0±0.5 vs. 11.2±0.5 fibers/mm) than CON (P&lt;0.05). In T1D, a deterioration from baseline to 5 years (n=151) was noted for ulnar and median MNCVs and SNCVs (e.g., ulnar MNCV: 57.4±0.4 vs. 56.3±0.3 m/s), malleolar VPT (0.76±0.07 vs. 1.12±0.12 µm), and HRV indices (e.g., standard deviation of normal RR intervals (SDNN): 69.3±2.4 vs. 62.0±2.1 ms; root mean square of successive differences (RMSSD): 42.8±2.3 vs. 34.7±2.0 ms) (all P&lt;0.05). Peroneal MNCV, sural SNCV, and TDT remained unchanged. The decline in MNCV was associated with an increase in HbA1c (e.g., median nerve: β=-0.316, P=0.004) and the deterioration in HRV with decreasing M-value (e.g., SDNN: β=0.246, P=0.041).
In conclusion, within the first 5-6 years of type 1 diabetes despite good glycemic control, the deterioration in median and ulnar MNCV was related to worsening HbA1c levels, while cardiac autonomic dysfunction progressed in relation to increasing insulin resistance.

Disclosure
G.J. Bönhof: None. A. Strom: None. O.P. Zaharia: None. J. Szendroedi: None. K. Muessig: None. M. Roden: Advisory Panel; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Poxel SA, Servier. Board Member; Self; Eli Lilly and Company. Research Support; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Sanofi. Speaker's Bureau; Self; Novo Nordisk Inc. D. Ziegler: Advisory Panel; Self; Allergan, TrigoCare. Consultant; Self; Mitsubishi Tanabe Pharma Corporation, Mylan, Novartis Pharmaceuticals Corporation, Wörwag. Speaker's Bureau; Self; Berlin-Chemie AG.

Funding
German Ministry of Culture and Science of the State of North Rhine-Westphalia; German Federal Ministry of Health; German Federal Ministry of Education and Research
33-OR: Intact Muscle Insulin Signaling in Patients with a Gain-of-Function LRP5 Mutation Despite Decreased Insulin Sensitivity
Authors: SABARATNAM RUGIVAN , LAUTERLEIN JENS-JACOB LINDEGAARD , NIELSEN MORTEN FROST , HOJLUND KURT 
Published: June 2019

Type 2 diabetes (T2D) is characterized by insulin resistance in skeletal muscle. GWAS studies have shown a link between T2D and Wnt signalling, and there is evidence that mutations in LRP5, a co-receptor in the canonical Wnt signaling pathway, may alter insulin sensitivity. Here, we investigated whether a gain-of-function (GoF) LRP5 mutation causing high bone mass is accompanied by changes in insulin sensitivity and abnormalities in the insulin signalling pathway in skeletal muscle. Skeletal muscle biopsies were obtained before and after a euglycemic-hyperinsulinemic clamp (3-h, 40 mU/min/m2) from 13 patients with GoF LRP5 mutation (T253I) and 14 age, and weight-matched controls. Protein abundance/phosphorylation of key mediators of insulin signalling to glucose transport and glycogen synthesis, such as Akt, TBC1D4, glycogen synthase kinase 3 (GSK3α and ß) and Erk1/2 were investigated. Insulin sensitivity measured as insulin-stimulated glucose disposal (Rd) was reduced (25%) in GoF LRP5 mutation carriers (p&lt;0.05). However, the insulin-stimulated phosphorylation of Akt at Ser473 and Thr308, TBC1D4 at Thr642 and Erk1/2 was fully intact in skeletal muscle of GoF LRP5 mutation carriers. Moreover, both the basal and the insulin-mediated inhibitory phosphorylation of GSK3 (α and ß) were unaltered in muscle of LRP5 mutation carriers compared with controls. To further investigate the molecular consequences of this GoF LRP5 mutation, we are currently investigating key regulators of Wnt signalling (activity of ß-catenin and CaMKII) and performing RNA-sequencing in muscle.
In conclusion, our data demonstrate that reduced insulin sensitivity in GoF LRP5 mutation carriers is not explained by abnormalities in the classical components of the insulin signalling cascade in skeletal muscle. This suggests that the GOF LRP5 mutation negatively affects insulin sensitivity by other mechanisms possible involving over-activation in Wnt signalling.

Disclosure
R. Sabaratnam: None. J. Lauterlein: None. M.F. Nielsen: None. K. Hojlund: None.
336-OR: An Online Insulin Pump and CGM Curriculum for Trainees: TeKnO T1D
Authors: MARKS BRYNN , WALDMAN GRETCHEN , STAFFORD DIANE E. , GARVEY KATHARINE , NAGLER JOSHUA , WOLFSDORF JOSEPH I. 
Published: June 2019

Background: A nationwide needs assessment survey identified suboptimal knowledge of insulin pumps and continuous glucose monitors (CGM) among pediatric endocrinology fellows.
Objectives: We developed and assessed the impact of an online, spaced education curriculum- Technology Knowledge Optimization in type 1 diabetes (TeKnO T1D)- on fellows’ knowledge of these technologies.
Methods: Endocrine fellows nationwide were invited to participate in this ongoing prospective, randomized cross over study. The pretest assessment included 20 multiple choice questions. A commercially available electronic spaced education platform (QStream) was used to deliver the material. The posttest assessment was identical and also included two questions assessing reactions to the curriculum. A linear multivariable regression was used to compare the deltas (pre-post) between the two groups adjusted for fellowship year and other covariates.
Results: Sixty-one fellows completed the pretest assessment. To date, 21 (68%) and 18 (60%) fellows have completed the pump and CGM curricula, respectively. Fellows who received the pump curriculum had a 41% greater mean improvement on the posttest questions focused on insulin pumps as compared to the fellow who completed the CGM curriculum (95% CI, 1.05, 1.88). Fellows who completed the CGM curriculum had a 70% greater mean improvement on the post-test questions focused on CGMs as compared to fellows who completed the pump curriculum (95% CI, 1.22, 1.37). The greatest improvements were seen in the following topics: interpretation of CGM trend arrows, pump management during exercise, managing pump disconnections, and unique features of different CGM systems. 97% of fellows agreed or strongly agreed that they enjoyed TeKnO T1D.
Conclusion: TeKnO T1D is an effective, enjoyable way to improve T1D technology knowledge across a large group of geographically dispersed fellows. This learning modality may also have applications for other healthcare professionals as well as patients and families.

Disclosure
B. Marks: None. G. Waldman: None. D.E. Stafford: None. K. Garvey: None. J. Nagler: None. J.I. Wolfsdorf: Other Relationship; Self; UpToDate, Xeris Pharmaceuticals, Inc.

Funding
Pediatric Endocrine Society; Boston Children's Hospital