bluuebunny/crossref_metadata_2025 huggingface dataset:
251-LB: Identification of Antibodies to Hybrid Insulin Peptides (HIPs) in T1D
Authors: GU YONG , WENZLAU JANET M. , MICHELS AARON W. , REWERS MARIAN , HASKINS KATIE , YU LIPING
Published: June 2019

Hybrid insulin peptides (HIPs) are unique post-translationally modified antigens whereby derivatives of proinsulin (PI) form peptide bonds with endogenous granule peptides. HIPs have been identified in mouse β-cell extracts and human islets by mass spectrometry. Diabetogenic CD4 T cell clones in NOD mice and CD4 T cells from T1D patients are stimulated by HIP ligands and imply that HIPs may also serve as epitopes for humoral immunity. To identify and characterize antibodies (Abs) targeting HIPs among T1D individuals and evaluate their utility as biomarkers, a sensitive and specific ECL-based HIP Ab assay was adapted from the hallmark ECL-IAA protocol. The key determinant for the HIP-Ab assay was design and implementation of a conformational HIP protein probe reflecting derivatives of the HIPs capable of eliciting an antigenic response in CD4 T cell lines from T1D individuals and NOD mice. Sera from a new onset T1D cohort using a PI/IAPP HIP probe showed the presence of anti-PI/IAPP Abs (32.2%, 58/180). However unexpected prevalence was measured among control (18.8%, 18/96) and T2D (25.9%, 29/112) sera samples even when the data was stratified according to HLA genotype. These HIP-Ab epitopes are both conformation and HIP-sequence specific as they are pre-absorbed only with refolded PI/IAPP, but not alternate or random HIPs, linear PI/IAPP HIPs or cyclic PI/IAPP peptides. In sera from prediabetic subjects from the DAISY cohort, 7 of 22 (31.8%) display a peak of HIP-Abs that appears prior to the peak of IAA autoAbs in most cases (87.5%, 7/8). Large number of individuals have PI/IAPP-specific Abs in their circulation, irrespective of T1D disease state or risk. Although HIP-Abs are pervasive and emerge early in the disease process in prospectively followed at risk prediabetic individuals, they are not predictive of disease. As the appearance of PI/IAPP HIP-Abs typically precedes IAAs, it is possible that Abs to other HIPs may serve as earlier biomarkers for ensuing disease.

Disclosure

Y. Gu: None. J.M. Wenzlau: None. A.W. Michels: Stock/Shareholder; Self; IM Therapeutics. M. Rewers: None. K. Haskins: None. L. Yu: None.
252-OR: Cardiorespiratory Fitness Predicts Lower Mortality Rates in Type 2 Diabetes
Authors: WILLS ANDREW C. , KNOWLER WILLIAM C. , CURTIS JEFFREY , OLAIYA MUIDEEN T.
Published: June 2019

Look AHEAD was a randomized multi-center clinical trial comparing effects of an intensive lifestyle intervention (ILI) with diabetes support and education (DSE) on CVD events in 5145 adults aged 45-76 years who were overweight or obese and had type 2 diabetes. After 10 years of follow-up, the ILI had no significant effect on the primary outcome of fatal or non-fatal CVD events or on all-cause or CVD mortality rates. We now present a secondary analysis of the effects of cardiovascular fitness at baseline on mortality in 4865 participants. Fitness was defined as the metabolic equivalence (METs) level achieved during maximal workload on a treadmill. For all participants, the mean (SD) were 7.20 (1.97) METs. The all-cause death rate was 14% lower per 1 MET greater fitness at baseline, adjusted for age, sex, race/ethnicity, CVD history, BMI and treatment group (HR 0.86 95% CI 0.80 -0.92). In this model, CVD history (HR 1.82 95% CI 1.42-2.35) but not ILI (HR 0.87 95% CI 0.71-1.07) significantly predicted mortality. Similarly, CVD mortality was 28% lower per 1 MET (HR 0.72, 95% CI 0.62-0.83). Fitness was similarly protective from the other CVD outcomes. The Figure shows all-cause mortality rates modeled by history of CVD, treatment group, age 60 years, and white men not on beta blockers because they were the largest population in the dataset.

In conclusion, fitness measured by maximal workload on a treadmill was highly predictive of CVD and mortality.

Disclosure

A.C. Wills: None. W.C. Knowler: None. J. Curtis: None. M.T. Olaiya: None.

Funding

National Institutes of Health
257-LB: Long-Term Effects of Birth Mode on Gut Microbiome Colonization
Authors: NIES LAURA DE , BUSI SUSHEEL BHANU , HABIER JANINE , WAMPACH LINDA , HEINTZ-BUSCHART ANNA , DE BEAUFORT CARINE , WILMES PAUL
Published: June 2019

Background: Caesarean section (CS) disrupts mother-to-neonate transmission of specific microbial strains, linked functional repertoires and immune system priming. We investigated whether the differences in microbial populations and functional potentials persist at 1 year of age.

Methodology: We performed high-resolution, quantitative metagenomic analyses of the gut microbiomes of 8 infants born by vaginal delivery (VD) (n=3) or by CS (n=5) at 1 year. Metagenomic sequencing reads were used to generate functional repertoire profiles based on the homology of the assembled genes/contigs with the KEGG database. Simultaneously, LPS from the infant stool samples was used to stimulate primary immune cells (dendritic cells) obtained from healthy human donors. An enzyme-linked immunosorbent assay for TNFα was used to determine concomitant levels of cytokines. Differential mOTU (metagenomic operational taxonomic unit) and KEGG pathway analyses were performed using the DESeq2 R package.

Results: Seven mOTUs including Anaerotruncus colihominis, [Ruminococcus obeum], and Subdoligranulum variabile demonstrated between 10- to 30- fold enrichment in the VD samples compared to the CS group (p < 0.05). Although ordination analyses did not indicate significant differences between the groups, the functional repertoire analysis found several KEGG pathways differentially enriched when comparing VD and CS samples. The CS samples exhibited enrichments in CD226 antigen, cell adhesion and vancomycin resistance pathways, while the VD group had an increased abundance of carbon metabolism and epithelial cell invasion pathways. Previously increased TNFα levels observed in VD samples right after birth compared to the CS group, were not observed at 1 year.

Conclusion: After 1-year, persistent differences in gut mOTUs were observed between CS and VD born infants. Whether the CD226 pathway upregulation relates to human auto-immune disease, requires further analysis and validation.

Disclosure

L. de Nies: None. S. Busi: None. J. Habier: None. L. Wampach: None. A. Heintz-Buschart: None. C. de Beaufort: Consultant; Self; Medtronic. P. Wilmes: None.

Funding

Fondation Andr? et Henriette Losch; National Research Fund; University of Luxembourg; Integrated BioBank of Luxembourg
257-OR: High Glucose Levels and Risk of Vascular Diseases—Observational Studies and Mendelian Randomization Studies of the General Population
Authors: EMANUELSSON FRIDA , MAROTT SARAH , TYBJAERG-HANSEN ANNE , NORDESTGAARD BØRGE GRØNNE , BENN MARIANNE
Published: June 2019

Aim: To investigate the observational and causal associations between glucose levels in the nondiabetic range and risk of vascular diseases in the general population.

Methods: We included 112,457 nondiabetic individuals from the Copenhagen City Heart Study and the Copenhagen General Population Study and assessed their prospective risk of retinopathy, neuropathy, diabetic nephropathy, peripheral arterial disease (PAD), and myocardial infarction (MI) as a function of increasing glucose levels. We used Mendelian randomization (MR) to assess a potential causal effect of high glucose levels on the vascular endpoints using genetic variants known to be associated with elevated glucose levels. We replicated our analyses in a 2-sample MR design using glucose data from the MAGIC consortium and vascular endpoint data from the UK Biobank.

Results: Observationally, glucose levels in the nondiabetic range and higher were associated with increased risks of retinopathy, neuropathy, diabetic nephropathy, PAD, and MI (p trend for all <0.001). In genetic, causal analyses risk ratios for a 1 mmol/L higher glucose were 2.01 (95% confidence interval: 1.18-3.41) for retinopathy, 2.15 (1.38-3.35) for neuropathy, 1.58 (1.04-2.40) for diabetic nephropathy, 1.19 (0.90-1.58) for PAD, and 1.49 (1.02-2.17) for MI. Summary level data from the MAGIC consortium and the UK Biobank gave similar results.

Conclusion: In this general population, glucose levels in the nondiabetic range and higher were prospectively associated with a high risk of retinopathy, neuropathy, diabetic nephropathy, PAD, and MI. These associations were causal for retinopathy, neuropathy, diabetic nephropathy, and MI, but causality could not be confirmed for PAD. These findings suggest that elevated glucose levels in the nondiabetic range is an important risk factor for microvascular diseases.

Disclosure

F. Emanuelsson: None. S. Marott: None. A. Tybjaerg-Hansen: None. B.G. Nordestgaard: None. M. Benn: None.

Funding

Danish Council for Independent Research
26-OR: Digenic FGFR1/KLB Variants Associated with Endocrine Specific FGF-21 Signaling Defects and Extreme Insulin Resistance
Authors: STONE STEPHEN I. , WEGNER DANIEL J. , WAMBACH JENNIFER A. , COLE F. SESSIONS , ORNITZ DAVID M. , URANO FUMIHIKO
Published: June 2019

Fibroblast growth factor 21 (FGF-21) is a hepatokine with multiple important metabolic functions. However, the role of FGF-21 in human insulin sensitivity remains unclear. We report an adolescent female with severe acanthosis nigricans and extremely elevated insulin levels (2446 uIU/mL). Due to her severe insulin resistance, we looked into the possibility that she might have a genetic syndrome of insulin resistance. Whole exome sequencing revealed that she carries 2 variants of unknown significance in genes critical for FGF-21 signaling, Fibroblast Growth Factor Receptor 1 (FGFR1) and beta-Klotho (KLB). FGFR1 and KLB are transmembrane co-factors that bind FGF-21 leading to downstream signaling. A bicistronic expression plasmid was created recapitulating the above variants found in our subject. These plasmids were transfected into L6 myoblasts, a cell line known to express low levels of endogenous FGF receptor. ERK phosphorylation (pERK), a major downstream signal of FGF-21, was measured after treatment with FGF-21. pERK was strongly upregulated in cells transfected with wild type FGFR1 and KLB (500,710 AU). However, variant FGFR1 and KLB greatly attenuated pERK (107,068 AU). Canonical FGF-2 treatment led to robust upregulation of pERK in both wild type and variant cell lines (374,968 AU and 441,654 AU respectively). Expression of Slc2a1 (Glut1) and Slc2a4 (Glut4) was also measured in response to FGF-21. Cells transfected with wild type FGFR1 and KLB led to a 1.7-fold increase in both Glut1 and Glut4 expression, whereas Glut1 and Glut4 expression was attenuated in cells transfected with the FGFR1 and KLB variants (1.2 and 0.7-fold respectively). These results suggest that the digenic FGFR1 and KLB variants found in our subject lead to an endocrine specific FGF-21 resistance with decreased GLUT1- and GLUT4-mediated glucose uptake. This helps explain our subject’s severe insulin resistance, and may represent a novel disorder related to FGF-21.

Disclosure

S.I. Stone: Research Support; Self; National Institute of Diabetes and Digestive and Kidney Diseases. Speaker's Bureau; Self; AbbVie Inc. D.J. Wegner: None. J.A. Wambach: None. F.S. Cole: None. D.M. Ornitz: None. F. Urano: Board Member; Self; Healthbeat. Research Support; Self; Aetas, Amylyx, JDRF, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health. Stock/Shareholder; Self; CytRx. Other Relationship; Self; Novus Biologicals.

Funding

National Institutes of Health (HL120002); Institute of Clinical and Translational Sciences/Children's Discovery Institute (UL1TR00448)
261-LB: Robotic Pancreas Kidney Transplantation for Type 1 Diabetic Patient with End-Stage Renal Disease: First Year Experience in Single Institute
Authors: DUK LEE SEUNG , CANTRELL LEIGH A. , OBERHOLZER JOSE
Published: June 2019

Background: Simultaneous pancreas kidney transplantation for type 1 diabetic patients with ESRD is a highly successful, but also very invasive surgery. Robotic assistance facilitates a minimally invasiveness approach to this technically intricate surgery. The present study aims to evaluate the feasibility and outcomes of the first year`s experience in robotic pancreas kidney transplantation (RPKT) for type 1 diabetic patients with ESRD.

Patients and Methods: From October 2017 to October 2018, we enrolled consecutive cases of RPKT at the University of Virginia. Operative and clinical features including graft and patient survivals were retrospectively analyzed. The organs were introduced via 6 cm Pfannenstil incision.

Results: Six patients (3 male/3 female) successfully underwent RPKT. Median age was 32 years old (range: 30-46years). Median pre-transplant body mass index (BMI) was 25.5 mg/kg2 (Range: 20.6-33.4 mg/kg2). Operative time and estimated blood loss were 593 min (473-733) and 300cc (150-400), respectively. Total cold and warm ischemic time were 436 min (242-585) for pancreas/725min (403-835) for kidney and 43 min (34-60) for pancreas/56min (44-69) for kidney, respectively. We created 5 enteric and 1 bladder drainage for transplant pancreas. There were no conversions to open. No major complications during surgery were observed. We reported one case of delayed graft function of kidney and two cases of rejection with complete recovery. We found one patient with a minor trocar wound dehiscence and revision of ureterocystostomy for stricture. There was one pancreas graft loss at postoperative 7 days due to graft vein thrombosis. No patient death was noted.

Conclusion: RPKT for type 1 diabetic patients with ESRD appears to be a safe and effective surgical procedure with potential advantages of wound complication, quality of vascular anastomosis, and shorter recovery with minimal incision compared to conventional open surgery.

Disclosure

S. Lee: None. L.A. Cantrell: None. J. Oberholzer: Employee; Self; CellTrans. Research Support; Self; JDRF. Stock/Shareholder; Self; Sigilon Therapeutics.
262-LB: Human Pancreas-Derived Multipotent Stromal Cells Secrete Islet Regenerative Stimuli
Authors: COOPER TYLER , MA JUN , BELL GILLIAN I. , LAJOIE GILLES , HESS DAVID A.
Published: June 2019

We recently demonstrated that intra-pancreatic (iPan) delivery of conditioned media (CM) generated by human bone marrow-derived multipotent stromal cells (hBM-MSC) can act as a biotherapeutic to reduce hyperglycemia in STZ-treated mice by stimulating endogenous islet regeneration. We have established analogous stromal cell lines derived from human islet cultures that grow adherent to plastic and possess multipotent mesodermal differentiation in vitro. Furthermore, CM generated by these human pancreas-derived MSC (hPanc-MSC) contained proangiogenic stimuli within extracellular vesicles (EVs) that accelerated blood vessel regeneration after injection into ischemic hindlimbs. Herein, we investigated whether iPan-injection of CM generated by hPanc-MSC could stimulate islet regeneration in STZ-treated mice. We hypothesized that islet regenerative stimuli would be harboured within EVs secreted by hPanc-MSC. iPan-injection of unfractioned CM (6µg) generated by hBM-MSC and hPanc-MSC equally reduced hyperglycemia in STZ-treated mice, compared to mice injected with media control (*p<0.05). To determine whether islet regenerative stimuli was harboured within EVs, ultrafiltration was used to separate CM into EV enriched (EV+) or depleted (EV-) subfractions. Enrichment of EVs was validated by nanoscale flow cytometry and atomic force microscopy, and the EV+ and EV- hPANC-MSC subfractions were compared for protein content by label-free mass spectrometry. Expression of typical EV-associated proteins (CD9, CD81, CD63) were exclusive to the EV+ fraction and putative islet regenerative factors were detected in both the EV- and EV+ subfractions C. Mice iPan-injected with either EV+ or EV- CM sub-fractions showed significantly reduced hyperglycemia compared to unconditioned media controls (*p<0.05). These initial studies provide a foundation to further explore the use the hPanc-MSC secretome or related small-molecule pharmaceuticals to enhance islet-regeneration in situ.

Disclosure

T. Cooper: None. J. Ma: None. G.I. Bell: None. G. Lajoie: None. D.A. Hess: None.
264-LB: Evaluation of Awareness about Insulin Injection Technique and Prevalence of Lipohypertrophy amongst Insulin-Taking Diabetics in Central India
Authors: SUNIL GUPTA SHLOK , CHAOJI SUNANDA , GUPTA SUNIL , GUPTA KAVITA , GATHE SACHIN
Published: June 2019

Background: Lipohypertrophy (LH) at insulin injection sites is a preventable complication in patients of diabetes mellitus taking insulin injection therapy. We evaluated the prevalence, contributing risk factors, and consequences of LH along with awareness regarding correct insulin injection technique amongst these diabetic patients.

Methodology: It is a cross sectional observational questionnaire-based study conducted at a tertiary health centre in Nagpur, India. 31 type 1 diabetics (T1DM) and 43 type 2 diabetics (T2DM) who are on insulin injection therapy for over one year, were randomly selected and presence of LH was evaluated through visual and palpation techniques. Demography, anthropometry, awareness of correct insulin injection technique were noted using a validated questionnaire. Their effect on LH was determined along with its correlation with complains of unexplained hypoglycaemia (UH).

Results: The prevalence of LH amongst T1DM is 48.39% and amongst T2DM is 18.60%. Prevalence was significantly higher in T1DM than in T2DM (p=0.0133). Young age (p=0.0042), Male gender (p=0.0011), decreased duration of diabetes (p=0.0281), improper rotation of the correct sites (p=0.0032) significantly increased the prevalence of LH. Occurrence of UH were significantly higher in patients with LH (p=0.0078). Independent risk factors for LH amongst T2DM include less age (p=0.0358), Male gender (p=0.0048) and improper rotation of sites (p=0.0071). No independent risk factors were found significant for LH amongst T1DM as well as no significance was observed for prevalence of LH with respect to needle reuse, number of injections taken per day and total daily dosage of insulin, which may be regarded to low sample size.

Conclusion: Improper rotation of injection sites, Male gender, younger age, lesser duration of diabetes and T1DM contribute to a higher prevalence of LH, which in turn will increase the risk of UH.

Disclosure

S.S. Gupta: None. S. Chaoji: None. S. Gupta: None. K. Gupta: None. S. Gathe: None.
27-LB: Alteration of the Gut Microbiome by Resveratrol Is Associated with Improvement in Function of Kidney in db/db Mice
Authors: DAFA DING , YIBING LU , TINGTING CAI , XIAOLONG YE , JIANHUA MAO
Published: June 2019

Oral administration of resveratrol is able to amelioration the progression of diabetic nephropathy, but the underlying cause is unknown. In this report, we demonstrated that resveratrol relieved a series of indicators of diabetic nephropathy. Oral administration of resveratrol was able to sequester negatively charged bacterial endotoxin in the gut, leading to 1) reduced plasma endotoxin levels, 2) resolved systemic inflammation and the Intestinal environment, and 3) lowered renal inflammation and improved renal function. Gut dysbiosis, characterized as an increment of Bacteroides, Alistipes, Turicibacter and Parabacteroides, and reduction of Enterococcus , was fully modulated by resveratrol, indicating that inflammatory-associated microbiome in the gut are critical for the metabolic endotoxemia. Furthermore, fecal bacteria transplant, derived from heathy resveratrol-treated mices, was sufficient to antagonize the renal dsyfunction and rebalance of gut microbiome in the recipient mice. These results indicate that resveratrol mediated changes in the gut microbiome may play an important role in the mechanism of action of resveratrol, which provide supporting evidence for the gut-kidney axis in DN.

Disclosure

D. Dafa: None. L. Yibing: None. C. Tingting: None. Y. Xiaolong: None. M. Jianhua: None.

Funding

Major Research and Development Project of Jiangsu (BE2016800); Nanjing Science and Technology Plan Project (BE201715015)
271-OR: Whole-Exome Sequence Identifies Potentially Functional Rare Variants in Bardet-Biedl Syndrome (BBS) Genes that Associate with Obesity in American Indians
Authors: DAY SAMANTHA E. , MULLER YUNHUA L. , KOBES SAYUKO , KIM HYE IN , VAN HOUT CRISTOPHER V. , GOSALIA NEHAL , YE BIN , SHULDINER ALAN R. , HANSON ROBERT L. , ALTOK CIGDEM KOROGLU , BOGARDUS CLIFTON , BAIER LESLIE J.
Published: June 2019

American Indians suffer from a high prevalence of obesity. Although their obesity is typically polygenic in nature, we are systematically analyzing known monogenic obesity genes in a population-based sample of Pima Indians. Previously, we have reported that 3% of this population carries functional mutations in MC4R. The goal of the current study is to determine whether variation in any of the 21 known BBS genes also contribute to obesity in this population. Analysis of whole exome sequence data from 6,809 Pima Indians (44.8% male; mean±SD age 34.8 ± 16.0 years; mean±SD maximum body mass index (BMI) 36.2 ± 8.8 kg/m2; 31.9% had T2D) identified 1,726 variants in the 21 BBS genes, of which 273 variants had a Combined Annotation Dependent Depletion (CADD) score >20 (top 1% most likely to be deleterious). Among these potentially deleterious variants, 32 were exclusive to, or enriched in, obese subjects (mean BMI for carriers >40). Five of the 32 variants were in the BBS9 gene and these were prioritized for functional studies. BBS mutations may impact obesity via leptin signaling, where BBS gene dysfunction has led to increased food intake. Therefore, in vitro luciferase reporter assays targeting STAT3 transcriptional activity were used to assess alterations in leptin signaling by the BBS variants. Analysis of a Thr549Ile (rs59252892) in BBS9 (mean BMI of Ile allele carriers was 41.3 ± 12.4 kg/m2 vs. 36.4 ± 8.8 kg/m2 for non-carriers, P=0.0008) showed that the Ile allele had a 28% increase in leptin signaling activity compared to the Thr allele (P=0.006, 5 experiments). Moreover, western blot analysis showed that the Ile allele had a 19% increase in STAT3 phosphorylation compared to the Thr allele. Comparable in vitro results were observed with a S545G, but not a K810E or L665F in BBS9.

In summary, whole exome sequencing in Pima Indians has identified potentially functional variants in BBS9 that may increase risk for obesity through leptin signaling.

Disclosure

S.E. Day: None. Y.L. Muller: None. S. Kobes: None. H. Kim: Employee; Self; Regeneron Pharmaceuticals. C.V. Van Hout: Employee; Self; Regeneron Pharmaceuticals. N. Gosalia: Employee; Self; Regeneron Pharmaceuticals. B. Ye: Employee; Self; Regeneron Pharmaceuticals. A.R. Shuldiner: Employee; Self; Regeneron Pharmaceuticals. Stock/Shareholder; Self; Rhythm Pharmaceuticals, Inc. R.L. Hanson: None. C. Koroglu Altok: None. C. Bogardus: None. L.J. Baier: None.
272-LB: The Micropeptide ORF60 Modulates Insulin Sensitivity
Authors: GUPTA NIKHIL , SHI YU , LI PING , CAO HAIMING
Published: June 2019

Although long non-coding RNAs (lncRNAs) are non-protein-coding transcripts by definition, recent studies have shown that a number of putative small open reading frames within lncRNAs are translated. The biological significance of these hidden polypeptides is still unclear, although studies have demonstrated functional significance for some polypeptides.

Recently, our lab discovered a novel micropeptide ORF60 encoded by a lncRNA. The micropeptide is conserved across species. Topology prediction indicates that the micropeptide is likely transmembrane. RNA transcript expression is nearly ubiquitous across tissues. In mice, ORF60 expression was found to be increased in the setting of high fat diet (HFD). Using CRISPR/Cas9 technology we developed ORF60 KO mice. In both mice feeding on normal chow diet and HFD, we observed impaired glucose tolerance in KO mice compared to wild type mice. ORF60 KO mice also demonstrated impaired response to insulin compared to WT mice on insulin tolerance testing (ITT), suggesting insulin resistance. Mouse tissue harvested under the condition of insulin infusion demonstrated insulin signaling attenuation in white adipose tissue (WAT) and brown adipose tissue (BAT). We are currently dissecting the molecular mechanism underlying ORF60’s influence on insulin signaling pathways.

In summary, our results demonstrate that the conserved micropeptide ORF60 regulates insulin sensitivity in WAT and BAT in mice. Micropeptides involved in diabetes may serve as novel drug targets.

Disclosure

N. Gupta: None. Y. Shi: None. P. Li: None. H. Cao: None.
272-OR: Metabolically Healthy Obesity Is Not So Healthy
Authors: COMMODORE-MENSAH YVONNE , LAZO MARIANA , TANG OLIVE , ECHOUFFO TCHEUGUI JUSTIN B. , NDUMELE CHIADI E. , NAMBI VIJAY , BALLANTYNE CHRISTIE M. , WANG DAN , SELVIN ELIZABETH
Published: June 2019

Background: It is unclear whether the “metabolically healthy” obese phenotype has low cardiovascular disease (CVD) risk. High-sensitivity cardiac troponin T (hs-cTnT), a biomarker of myocardial damage, characterizes subclinical CVD risk. We categorized obesity phenotypes and studied their cross-sectional and prospective associations with subclinical and clinical CVD.

Methods: We included 11,844 adults in the Atherosclerosis Risk in Communities (ARIC) Study. We used ATP III metabolic syndrome criteria and BMI to define obesity phenotypes as metabolically healthy non-obese (MHNO), metabolically unhealthy non-obese (MUNO), metabolically healthy obese (MHO), and metabolically unhealthy obese (MUO).

Results: At baseline (1990-1992), mean age was 56 years, 56% were female and 23% black. The MHO group was more likely to have detectable hs-cTnT (≥ 6 ng/L) than the MHNO (OR 1.23, 1.02-1.49). During over 26 years of follow-up, 3,560 participants developed CVD. The metabolically unhealthy and obese phenotypes had higher CVD risk than MHNO (Table). Detectable hs-cTnT was associated with higher CVD risk in all phenotypes; MHO had CVD risk, which was intermediate between MHNO and MUO regardless of hs-cTnT (Table).

Conclusion: Our results show that metabolically “healthy” obesity is associated with substantial excess burden of subclinical and clinical CVD. Lifestyle modification among obese adults, regardless of metabolic status, is vital to prevent CVD.

Disclosure

Y. Commodore-Mensah: None. M. Lazo: None. O. Tang: None. J.B. Echouffo Tcheugui: None. C.E. Ndumele: None. V. Nambi: Other Relationship; Self; Dynamed, Merck & Co., Inc., Roche Diagnostic USA. C.M. Ballantyne: Consultant; Self; Abbott, Akcea Therapeutics, Amarin Corporation, Amgen Inc., AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Eli Lilly and Company, Esperion, Gilead Sciences, Inc., Matinas BioPharma, Merck & Co., Inc., Novartis Pharmaceuticals Corporation, Novo Nordisk Inc., Regeneron Pharmaceuticals, Roche Diagnostic USA, Sanofi. Research Support; Self; Abbott, Akcea Therapeutics, Amarin Corporation, Amgen Inc., Esperion, Novartis Pharmaceuticals Corporation, Regeneron Pharmaceuticals, Roche Diagnostic USA, Sanofi. Other Relationship; Self; Roche Diagnostic USA. D. Wang: None. E. Selvin: None.

Funding

National Institutes of Health (5KL2TR001077-05)
273-OR: Intrapancreatic Adipose Tissue of Nondiabetic and Type 2 Diabetic Subjects and Relationships with Islet Morphometry
Authors: SULEIMAN MARA , LUCA CARMELA DE , SIMONE PAOLO DE , BOGGI UGO , GRANO FRANCESCA , BUGLIANI MARCO , MARCHETTI PIERO , MARSELLI LORELLA
Published: June 2019

Little information is available on adipose tissue features in the human pancreas (AdTP). In the present study we performed morphologic and morphometric studies on AdTP of 9 overweight/obese nondiabetic (ND, age: 65±5 years; 4M/5F; BMI: 31.3±0.4 Kg/m2, mean±SEM) and 13 matched type 2 diabetic (T2D, age: 70±2 years, 8M/5F; BMI: 30.0±0.8 kg/m2) organ donors. Adipocytes were identified and quantified on a fluorescence DM5500 Leica microscope, using the MetaMorph v 1.8.0 software with a tresholded mask applied in exclusive mode. Macrophages were identified by immunohistochemistry using anti-CD68 (Dako) antibody. Assessment of insulin (Abcam Ab) and glucagon (Sigma Ab) positive cells was also performed. The number of adipocytes trended higher (+32.8%) in T2D (8.9±1.4/mm2) compared to ND (6.7±2.1/mm2) and their size was greater in T2D (9,319±1,028 μm2) than ND (5,639±639 μm2, p=0.01). AdTP area in relation to acinar tissue was 90.4% higher (p=0.07) in T2D (7.6±1.3%) than ND (3.9±1.5%). The amount of adipocytes with adjacent ≥ 3 CD68+ cells was higher (p=0.048) in T2D (31/281, 11%) than ND (7/147, 5%). Insulin positive area (T2D: 0.49±0.06%; ND: 0.55±0.09%) as well as glucagon positive area (T2D: 0.24±0.03%; ND: 0.32±0.07%) did not differ significantly between the two groups. Adipocyte size was negatively associated with islet number in ND+T2D (p=0.06) and positively with insulin area in ND (p=0.058); there was a significant, positive relationship between the amount of adipocytes and the number of islets in T2D (p=0.017).

In conclusion, we found significant differences in the AdTP of T2D as compared to ND, including increased adypocite size and presence of adipocyte adjacent CD68+ cells; some of these parameters correlated with islet morphometric indices.

Disclosure

M. Suleiman: None. C. De Luca: None. P. De Simone: None. U. Boggi: None. F. Grano: None. M. Bugliani: None. P. Marchetti: None. L. Marselli: None.

Funding

Innovative Medicines Initiative 2 Joint Undertaking (115881); European Union; European Federation of Pharmaceutical Industries and Associations; Swiss State Secretariat for Education, Research and Innovation (16.0097-2)
275-OR: Higher Liver Fat and Whole-Body Insulin Sensitivity in Newly Diagnosed Type 2 Diabetes Patients with a Variant in Transmembrane 6 Superfamily Member 2 Protein
Authors: BODIS KALMAN , SZENDROEDI JULIA , AHLERS JULIANE MARIA , KARUSHEVA YANISLAVA , ZAHARIA OANA P. , ANTONIOU SOFIA , KNEBEL BIRGIT , KUPRIYANOVA YULIYA , HWANG JONG-HEE , BURKART VOLKER , MUESSIG KARSTEN , AL-HASANI HADI , RODEN MICHAEL , MARKGRAF DANIEL F.
Published: June 2019

Knockdown of the gene encoding transmembrane 6 superfamily member 2 protein (TM6SF2) in mice increases hepatocellular lipid content (HCL) and decreases very low-density lipoprotein secretion. Humans with TM6SF2 mutations are at higher risk of nonalcoholic fatty liver disease, but lower risk of cardiovascular disease. The relevance of TM6SF2 mutations for HCL and insulin sensitivity in type 2 diabetes is less clear. This study aimed at elucidating the relevance of the E167K (SNP rs58542926) mutation in the gene encoding TM6SF2 for HCL, hepatic insulin sensitivity and whole-body insulin sensitivity in type 2 diabetes.

We compared male patients with recently diagnosed type 2 diabetes with (n=16) or without (n=16) E167K mutation, matched for age and BMI (50±1 vs. 48±3 years; 32±1 vs. 32±2 kg/m2) from the German Diabetes Study. Whole-body and hepatic insulin sensitivity were assessed by Botnia-clamp tests with [6,6-2H2]glucose, HCL by 1H magnetic resonance spectroscopy and hepatic fibrosis by the fibrosis-4 index (FIB4).

Patients with and without TM6SF2 mutation had comparable levels of HbA1c (6.2±0.2 vs. 5.9±0.1%) and serum triglycerides (136±14 vs. 149±14 mg/dl). Carriers of TM6SF2 mutation had higher HCL (9.0±1.9 vs. 4.6±0.4%, p<0.05) and whole-body insulin sensitivity (7.1±0.6 vs. 5.4±0.2 mg/(kg*min), p<0.05), but similar hepatic insulin sensitivity (1.6±0.1 vs. 1.6±0.1 mg/(kg*min)) and fibrosis score (1.2±0.1 vs. 1.0±0.1).

In conclusion, the E167K mutation of the TM6SF2 gene dissociates liver steatosis from whole-body insulin sensitivity. This may result from favored trapping of triglycerides within the liver, which would in turn protect peripheral tissues from insulin resistance induced by liver-derived lipids.

Disclosure

K. Bodis: None. J. Szendroedi: None. J.M. Ahlers: None. Y. Karusheva: None. O.P. Zaharia: None. S. Antoniou: None. B. Knebel: None. Y. Kupriyanova: None. J. Hwang: None. V. Burkart: None. K. Muessig: None. H. Al-Hasani: 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.F. Markgraf: Research Support; Self; Sanofi.
28-OR: Genetic Landscape of Diabetes in the Swiss Population
Authors: SCHWITZGEBEL VALERIE , BLOUIN JEAN-LOUIS , SANTONI FEDERICO
Published: June 2019

Background/Introduction: Diabetes represents a heterogeneous group of metabolic disorders and monogenic diabetes is thought to affect 1- 5% of the diabetes population. Monogenic diabetes causes neonatal diabetes (ND), maturity-onset diabetes of the young (MODY), and syndromic diabetes. The identification of the gene underlying diabetes remains crucial. In many cases, it will instruct a personalized treatment regime, thus improving metabolic control. The aim of the study was to identify mutations causing genetic diabetes.

Methods: Inclusion criteria for testing were ND, autoantibody negative type 1 diabetes (T1D), presumed MODY, type 2 diabetes (T2D) diagnosed before the age of forty-five without metabolic features and syndromic diabetes. Haloplex technology was used to perform targeted next-generation sequencing (NGS) for 323 genes known to be involved in glucose homeostasis.

Results: We have analyzed 234 diabetic individuals (137 females, 97 males) living in Switzerland by NGS. The mean age at diabetes diagnosis was 27.7 ±15.4 years. We identified variants in the MODY genes 1-13 in 41% (95/234) of the patients, of which 7 had variants in two, and one in three genes. The most frequent variants involved GCK in 58% (55/95), followed by HNF1A 19%, HNF4A 7%, ABCC8 6%, HNF1B/KCNJ11/KLF11 3%, NEUROD1 2% and PAX4 1%. When mapped to age at diabetes onset, MODY variants were most frequent in the neonatal group (50%, 2/4), and declined with age to reach 36% (13/36) in the >45 year group. In the group with variants in other than MODY genes (n=111) diabetes onset was 30 ±16 years; in 39% of the individuals we identified one genetic variant with a maximum of up to six in 1.8% of the probands.

Conclusion: NGS is now a feasible method to diagnose monogenic diabetes, as we identified variants in MODY genes in 41% of probands in our cohort. We detected potential digenic and oligogenic diabetes forms in 61% of the probands with variants in other than MODY genes. The discovery of additional diabetes genes may lead to novel, rational treatment strategies.

Disclosure

V. Schwitzgebel: None. J. Blouin: None. F. Santoni: None.

Funding

Swiss National Science Foundation
283-OR: Structured Nurse-Led Intervention following Ambulance Call Out for Hypoglycemia Reduces Cardiovascular Mortality in Type 2 Diabetes
Authors: PEARSON SAM , KULAVARASALINGAM KAVITA , BAXTER PAUL , MITCHELL-GEARS AMELIA K. , WHITTAM BEVERLEY ANN , AJJAN RAMZI
Published: June 2019

Aims: The aim of this work is to provide in depth characterization of patients with type 2 diabetes (T2D) with severe hypoglycemia and assess the role of structured nurse-led intervention on clinical outcome.

Methods: Individuals with T2D and severe hypoglycemia requiring ambulance services assistance were recruited into the study. Participants were randomized into standard arm (n=64), managed using existing pathways, or intervention arm (n=60), receiving nurse-led support for a period of 90 days. A third group did not wish to participate in the interventional study but consented for their data to be collected and were analyzed separately (observational arm; n=36).

Results: Of the 160 participants, 88 were men and median age was 77.0 (IQR 69.3-82.0) years. Mean HbA1c±SD was 59.7±16.1 mmol/mol and capillary glucose levels at ambulance crew arrival was 2.2±0.75 mmol/L. HbA1c, smoking status, age and presenting glucose were similar in the standard and intervention arms. 9 participants withdrew or lost to follow-up and were excluded from analysis. Individuals were followed-up for a median of 1350 days (IQR 870-1627) and mortality occurred in 67 participants (44.4%), with cardiovascular events as the leading cause (34%) followed by infections (27%). The number of deaths in the intervention arm was 17 (32%) compared with 31 (50%) in conventionally managed individuals (p=0.047), mainly due to a reduction in cardiovascular mortality [1 (2%) and 11 (18%), respectively; p<0.01]. Individuals in the observational and standard arms had similar mortality rate.

Discussion: This pilot study demonstrates that cardiovascular events and infections are the main cause of mortality following severe hypoglycemia in the community requiring ambulance services assistance. A structured nurse-led intervention for 3 months reduces mortality primarily due to a decrease in cardiovascular events.

Disclosure

S. Pearson: Speaker's Bureau; Self; Novo Nordisk Inc. K. Kulavarasalingam: None. P. Baxter: None. A.K. Mitchell-Gears: Other Relationship; Self; Heart Research UK. B.A. Whittam: None. R. Ajjan: Consultant; Self; Abbott. Research Support; Self; Abbott, LifeScan, Inc.
289-OR: Aerobic Exercise Counteracts Mitochondrial Dysfunction in the Insulin Resistant Brain
Authors: RUEGSEGGER GREGORY , VANDERBOOM PATRICK M. , DASARI SURENDRA , KLAUS KATHERINE , NAIR K. SREEKUMARAN
Published: June 2019

Insulin resistance increases the risk for Alzheimer’s disease and other dementias; however, the underpinning mechanisms for this increased risk remain to be fully defined. Insulin resistance impairs mitochondrial oxidative metabolism and increases reactive oxygen species (ROS) in skeletal muscle, both of which are counteracted by aerobic exercise (AE). Therefore, we considered whether similar events occur in the brain, specifically in regions such as cerebrum and hippocampus which are rich in insulin receptors and mitochondria. We determined whether AE would counteract mitochondrial deficits in hippocampal and cerebral mitochondrial function in high fat diet (HFD) induced insulin resistance. Four weeks of HFD in mice induced hippocampal insulin resistance which was corrected by AE. HFD decreased ATP production and increased ROS emission in isolated cerebral mitochondria, which were rescued with AE. Impairments in mitochondrial function with HFD were paralleled by reductions in mtDNA copy number, mRNA expression of mitochondrial genes, and oxidative enzyme activities (e.g., citrate synthase, COX) in the hippocampus, which were corrected by AE. We also observed excessive mitochondrial fission in the hippocampus following HFD, which was prevented by AE. Proteomic analysis of the hippocampus showed that HFD led to oxidative post-translational modifications (PTMs) of mitochondrial proteins; however, this increase in oxidative PTMs to mitochondrial proteins with HFD was almost completely reversed by AE. Further, intranasal administration of the insulin receptor antagonist S961 prevented AE-induced improvements in ATP production and ROS emission in isolated cerebral mitochondrial during HFD, demonstrating that AE enhances cerebral mitochondrial function during HFD by resensitizing brain insulin signaling.

Disclosure

G. Ruegsegger: None. P.M. Vanderboom: None. S. Dasari: Consultant; Self; The Binding Site. K. Klaus: None. K. Nair: None.
290-LB: Repeated Neonatal Oral Sucrose Treatment Affects Growth and Insulin-Like Growth Factor-1 in Mice
Authors: CONTRERAS CYNTHIA YAMILKA RAMIREZ , MUSSAI EI-XIA , WIEDEMAN ALEJANDRA M. , MEHRAN ARYA , SALEHZADEH MELODY , BOONPATTRAWONG NICHA , RANGER MANON , SOMA KIRAN K. , HOLSTI LIISA , DEVLIN ANGELA M.
Published: June 2019

Premature infants (<;37 weeks of gestation) often require hospitalization in the neonatal intensive care unit, where they may experience painful procedures due to medical care. Oral sucrose is the non-pharmacological standard of care for minor procedural pain relief; 0.5-2mL of a 24% sucrose solution is administered orally prior to each painful procedure. Throughout hospitalization infants can be exposed to high and cumulative volumes of sucrose; however, little is known about the long-term effects of oral sucrose on growth and development. The objective of this study was to determine the long-term effects of repeated neonatal oral sucrose treatment on growth, adiposity, and glucose homeostasis. Neonatal female and male mice (C57BL/6J) were randomly assigned to one of five treatment groups (n=6-10 mice/group/sex): sham (only handled), sterile water, sucrose, fructose, or glucose. Pups were treated 10 times/day for the first six days of life with 0.2g/kg weight of respective treatments (1-4 μl/dose; 24% solutions) orally to mimic what is given to preterm babies. At 3 weeks (weeks), mice were weaned onto a control diet and fed until euthanasia, at 16 weeks. There were no effects of neonatal oral sucrose treatment on glucose and insulin tolerance, and glucose stimulated insulin secretion at weaning or during adulthood. Female sucrose-treated compared to water-treated mice gained less weight (15.4±1.60 vs. 17.4±1.90g), had smaller tibias (15.9± 0.35 vs. 16.67±0.24mm), and lower serum insulin-like growth factor 1 (IGF-1) (378± 98.0 vs. 661±196ng/ml) at 16 weeks. No differences in growth were found in males. However, IGF-1 levels were lower in sucrose-treated vs. sham male mice (401± 143 vs. 641±165ng/ml). Our findings suggest that repetitive neonatal sucrose treatment may affect growth in females through an IGF-1-dependent pathway.

Disclosure

C.Y. Ramirez Contreras: None. E. Mussai: None. A.M. Wiedeman: None. A. Mehran: None. M. Salehzadeh: None. N. Boonpattrawong: None. M. Ranger: None. K.K. Soma: None. L. Holsti: Other Relationship; Self; Provincial Health Services Association. A.M. Devlin: None.

Funding

British Columbia Children's Hospital Research Institute
290-OR: High-Intensity Interval Training and Moderate-Intensity Continuous Training Induce Similar Modifications to Factors Regulating Skeletal Muscle Lipolysis
Authors: RYAN BENJAMIN J. , SCHLEH MICHAEL W. , VARSHNEY PALLAVI , LUDZKI ALISON , GILLEN JENNA B. , FOUG KATIE , CARR BENJAMIN D. , HOROWITZ JEFFREY F.
Published: June 2019

Abnormalities in muscle lipid metabolism in obesity have been linked to insulin resistance. Exercise training alters skeletal muscle lipid abundance and localization, but the direct effects of exercise training (independent of weight loss) on skeletal muscle lipolytic proteins are not clearly understood, and data are lacking regarding influence of training intensity on factors regulating muscle lipid metabolism in obesity. Our aim was to examine the effects of high-intensity interval training (HIIT) vs. moderate-intensity continuous training (MICT) on skeletal muscle lipolytic proteins in obese humans. Eighteen sedentary, obese adults completed 12 weeks (4 sessions weekly) of either HIIT (10 x 1 min at 90% HRmax, 1 min recovery; n=8) or MICT (45 min at 70% HRmax; n=10). Muscle biopsies (vastus lateralis) were collected before and after training. Subjects maintained body weight and fat mass and the post-training biopsy occurred 3 days after the final exercise session. Both exercise training programs increased aerobic capacity (VO2max) by ~10% (p=0.003), with no significant difference between HIIT and MICT. In muscle samples, hormone-sensitive lipase (HSL) protein abundance increased ~2-fold after training in HIIT (P<0.05), but not MICT. Training did not affect muscle abundance of adipose triglyceride lipase (ATGL) in either group but interestingly, the abundance of both CGI-58 (a positive regulator of ATGL lipolytic activity) and G0S2 (a negative regulator of ATGL lipolytic activity) increased ~15-20% after training (p<0.02), with no difference between HIIT and MICT.

In summary, HIIT and MICT result in similar modifications to lipolytic proteins in skeletal muscle, although our findings suggest that HIIT may be a more potent stimulus for increasing HSL abundance. Future work is needed to determine if the concomitant increases in protein abundance of CGI-58 and G0S2 with training may improve lipid handling in obesity.

Disclosure

B.J. Ryan: None. M.W. Schleh: None. P. Varshney: None. A. Ludzki: None. J.B. Gillen: None. K. Foug: None. B.D. Carr: None. J.F. Horowitz: Research Support; Self; American Diabetes Association.

Funding

National Institutes of Health (R01DK077966, P30DK089503, T32DK007245); Canadian Institutes of Health Research (DFS146190)
295-OR: User Acceptance of the Eversense® CGM System in a Home Use Setting: The House Study
Authors: LONDAHL MAGNUS , LANDIN-OLSSON MONA , ATTVALL STIG
Published: June 2019

Background: The Eversense® CGM System is the first implantable CGM available for long-term continuous glucose measurement with a single sensor. The small sensor is indicated for subcutaneous wear in the upper arm. This study was conducted to assess usability and preference for arm or abdominal sensor wear.

Methods: HOUSE was a prospective, single-arm, two-center study conducted in two sequential 90-day periods in subjects with T1D. During the first period, subjects were inserted with 2 sensors: one in upper arm and one in abdomen. After 90 days, both sensors were removed and subjects chose either the upper arm or abdominal location for the next sensor. Study visits occurred every 30 d up to 180 d for device downloads/safety assessment. At the 90-day and 180-day visit, subjects filled out a custom-designed questionnaire on their experience with the CGM system.

Results: Eighteen subjects participated (83% male, age 40±12.5 years, BMI 25.6±4.4kg/m2, 23 years from diabetes diagnosis; mean±SD). Sixty-one percent were on MDI and 89% had prior CGM experience. Two subjects withdrew consent due to personal reasons and 100% of the remaining sensors lasted each 90-day cycle. Ninety-four percent preferred to wear the sensor in the upper arm with 72% highly agreed/agreed that the transmitter was comfortable on the arm and 23% highly agreed/agreed that the transmitter was comfortable on the abdomen. The majority of subjects reported that sensor insertion was painless (69%) and that they were able to wear the transmitter in their everyday setting (94%). Product likeability scores were high: 94% on displaying readings on a smartphone and 75% on predicted hypo-/hyperglycemic alerts. The median wear time was 23.4 hours/day and 88% of subjects reported reviewing their CGM glucose values at least every other hour while awake.

Conclusion: In a home-use setting with experienced CGM users, subjects preferred wearing the Eversense on the arm compared to the abdomen. The system was rated high for usability on either the arm or abdomen.

Disclosure

M. Londahl: Advisory Panel; Self; AstraZeneca, Boehringer Ingelheim International GmbH, Novo Nordisk A/S, Sanofi. Speaker's Bureau; Self; AstraZeneca, Boehringer Ingelheim International GmbH, Novo Nordisk A/S, Sanofi. M. Landin-Olsson: None. S. Attvall: None.