bluuebunny/crossref_metadata_2025 huggingface dataset:
28-LB: Diabetic Retinopathy Is an Additional Risk Factor for Hemodialysis Induction and Mortality in Type 2 Diabetic Patients
Authors: MORIYA TATSUMI , MATSUBARA MADOKA , OUCHI MOTOSHI
Published: June 2019

High urinary albumin excretion (UAE) and low glomerular filtration rate (GFR) are risk factors for progressive renal function loss in type 2 diabetic patients. In addition, diabetic retinopathy (DR) is also a risk factor for progressive renal function decline in microalbuminuric type 2 diabetic patients. We aimed to elucidate factors, including DR, associated with more severe situation, hemodialysis (HD) induction or mortality in 325 type 2 diabetic patients whose GFRs were measured by the iohexol injection in 1995-1997. Two-hundred and sixty were followed for >5 years (mean follow-up period, 16±6 years). The UAE levels were classified as normoalbuminuria (NA; n=130), microalbuminuria (MA, n=96), and macroalbuminuria (ON; n=34). Patients were also divided into the normofiltration (NF; n=213) and low filtration (LF; n=47) groups according to the age-appropriate GFR range. One hundred patients had DR and 119 had hypertension. During the study period, 25 patients received HD induction and 39 died. The HD induction and mortality rates in the LF, ON, and DR groups were significantly higher than those in the NF, NA and MA, and no-DR groups, respectively. A Kaplan-Meier analysis revealed LF (log rank p<0.001), ON (p<0.001) and DR (p<0.001) were associated with a significantly higher risk of HD induction than NF, NA and MA and no-DR, respectively. A Kaplan-Meier analysis revealed that LF (p<0.001), ON (p<0.001) and DR (p=0.008) were associated with a significantly higher risk of mortality than NF, NA and MA and no DR, respectively. A stepwise regression analysis revealed LF (F=28.0, p=0.000), ON (F=34.4, p=0.000) and DR (F=7.9, p=0.005) affected the HD induction, but hypertension did not (F=0.2, p=0.6). LF (F=13.2, p=0.000), ON (F=4.4, p=0.04) and DR (F=6.4, p=0.01) affected the mortality, but hypertension did not (F=1.6, p=0.2). DR should also be considered as a risk factor to no small extent for HD induction and mortality in addition to UAE and GFR in type 2 diabetic patients.

Disclosure

T. Moriya: None. M. Matsubara: None. M. Ouchi: None.
280-LB: Role of A1 and A3 Adenosine Receptors in Whole Body Glucose Metabolism
Authors: JAIN SHANU , TOSH DILIP K. , REITMAN MARC , JACOBSON KENNETH A.
Published: June 2019

A1 and A3 adenosine receptors (ARs) belong to the group of GPCRs that primarily couple to Gi/o protein and mediate adenosine’s biological actions. ARs are widely expressed in the body and regulate various metabolic functions. We generated mice lacking two ARs, Adora1-/-; Adora3-/- (double knockout [A1A3DKO]) to enable investigation of the role of Gi-coupled ARs on glucose metabolism. The Adora1-/- (A1KO), Adora3-/- (A3KO) and WT mice generated as littermates were used as controls. A1A3DKO mice showed no difference in body weight, glucose tolerance, insulin sensitivity, fed blood glucose and plasma insulin levels on regular chow (RC). Fasting blood glucose levels were reduced in A1KO and A1A3DKO mice compared to WT mice. No difference in body weight was observed between A1A3DKO and control groups on high fat diet (HFD). A1KO mice showed improved glucose tolerance compared to A3KO mice without any difference in insulin sensitivity. No difference in plasma insulin levels was observed between the groups. Fasting blood glucose levels were reduced in A1A3DKO mice and A1KO mice compared to WT and A3KO mice, respectively. We investigated the contribution of liver to the observed decrease in fasting blood glucose levels. Pyruvate tolerance test revealed decreased glucose output from the liver in A1KO and A1A3DKO mice. Expression levels of hepatic genes involved in gluconeogenesis, lipid uptake, lipogenesis and inflammation were reduced in A1KO and A1A3DKO mice. Activation of A1AR by specific agonist (MRS7469) increased the phosphorylation of c-Jun N-terminal Kinases (JNK). Increased hepatic glucose production because of Gi signaling-mediated JNK activation is well documented. Taken together, our data support the concept that A1AR antagonists may prove valuable in enhancing whole-body glucose homeostasis by improving hepatic metabolism and could be beneficial in diabetes treatment.

Disclosure

S. Jain: None. D.K. Tosh: None. M. Reitman: None. K.A. Jacobson: None.
281-OR: Directness and Sustainability of rtCGM Effects on Hypoglycemia: A Secondary Analysis of the Hypode Study
Authors: EHRMANN DOMINIC , HEINEMANN LUTZ , FRECKMANN GUIDO , WALDENMAIER DELIA , HERMANNS NORBERT
Published: June 2019

The HypoDE study demonstrated that real-time continuous glucose monitoring (rtCGM) can reduce biochemical as well as clinical hypoglycemia in people with hypoglycemia problems. However, it is not clear how quick a reduction of low glucose values via rtCGM occurs and how sustainable this is. We analyzed the rtCGM group of the HypoDE study and compared the 4-week baseline phase with masked rtCGM to three 4-week intervals within the therapy/outcome phase (T/O phase) with open rtCGM (Dexcom G5). Only participants with no recent experience with CGM were included. At baseline, 6.5±6.2% of the glucose values of the 75 participants were below 70 mg/dl and 2.5±3.3% were below 54 mg/dl. In the first week, these percentages dropped significantly to 2.9±0.4%, respectively 0.7±1.3%. For the remainder of the T/O phase, these percentages stayed stable (not crossing the lower 95% CI interval of the baseline values; see Figure). Time-in-Range (TIR) improved significantly in the first 12 weeks and then started to drop to baseline level. rtCGM results in an immediate and significant reduction of level 1 (< 70 mg/dl) and level 2 (< 54 mg/dl) hypoglycemic values. This initial effect can be sustained throughout the entire study period without wear off. The avoidance of hypoglycemia was not associated with a deterioration of TIR.

Disclosure

D. Ehrmann: Advisory Panel; Self; Medtronic. Speaker's Bureau; Self; Abbott, Berlin-Chemie AG, Dexcom, Inc. L. Heinemann: Consultant; Self; Roche Diabetes Care. Research Support; Self; Dexcom, Inc. Stock/Shareholder; Self; Profil Institute for Metabolic Research, ProSciento. G. Freckmann: Advisory Panel; Self; Abbott, Novo Nordisk Pharma GmbH, Sensile Medical AG. Consultant; Self; Metronom Health Europe, Roche Diabetes Care, Sanofi. Speaker's Bureau; Self; Ascensia Diabetes Care, Dexcom, Inc., Ypsomed AG. D. Waldenmaier: None. N. Hermanns: Advisory Panel; Self; Abbott. Board Member; Self; Eli Lilly and Company. Research Support; Self; Abbott, Berlin-Chemie AG, Dexcom, Inc., Roche Diabetes Care. Speaker's Bureau; Self; Berlin-Chemie AG, Dexcom, Inc., Novo Nordisk A/S.

Funding

Dexcom, Inc.
284-LB: Quantifying and Localizing the Mitochondrial Proteome
Authors: WU YIBO
Published: June 2019

Mitochondria are essential organelles that regulate a wide range of cellular functions. Here, we have developed data-independent acquisition mass spectrometry (DIA-MS) strategy and have quantified 3648 proteins in 114 proteomes collected from the BXD genetic reference populations in five tissues: liver, heart, brain, brown adipose tissue (BAT) and quadriceps. Protein expression varied more significantly across tissues than across genetic backgrounds. Although we observed broad correlation between transcript and protein expression, genes in the same pathway may have striking difference in regulation on transcript and protein levels, for example the genes in the electron transport chain. We compared two fractions from each tissue, mitochondria and total tissue. By comparing these two fractions, we identified several dozen proteins as mitochondrial proteins, which have not been reported in the literature. Among them, we validated three proteins in mitochondria (MTAP, SOAT2 and IMPDH2), and one protein in the mitochondria-associated membrane (ABCC6). Our results have revealed the disrupted protein composition of complex V of the electron transport chain on protein level, but not on the transcript level. Furthermore, we confirmed UCP1 protein level as a better predictor for cold test response compared with its transcript level. These findings demonstrate the synergy of proteomics and genomics to study complex metabolic processes, and they provide a resource for further discovery and analysis of proteoforms and protein localization.

Disclosure

Y. Wu: None.
286-LB: The Difference in Metabolic Profiles between Liver Cirrhosis Patients and Healthy Control before Liver Cirrhosis Development: A Prospective, Korean Cancer Prevention Study-II Cohort Study
Authors: YOO HYE JIN , KIM MINJOO , HAN YOUNGMIN , YOONJEONG CHOI , LEE JONG HO
Published: June 2019

Objectives: In the prospective Korean Cancer Prevention Study-II (KCPS-II), we investigated the application of metabolomics to differentiate subjects with incident liver cirrhosis (LC group) from subjects who remained free of LC (control group) during a mean follow-up period of 7 years, with the aim of identifying valuable biomarkers for LC.

Methods: Metabolic alterations were investigated using baseline serum samples from 94 subjects with incident LC and 180 age- and sex-matched LC-free subjects via ultra-performance liquid chromatography (UPLC)-linear-trap quardrupole (LTQ)-Orbitrap mass spectrometry (MS).

Results: A total of 46 metabolites were identified through the metabolic analysis. Among them, 11 metabolite levels were significantly higher, and 18 metabolite levels were significantly lower in the LC group than in the control group. 9 metabolic pathways, including glyoxylate and dicarboxylate metabolism, amino acid metabolism, fatty acid metabolism, linoleic acid metabolism, α-linolenic acid metabolism, and arachidonic acid metabolism, showed significant differences between the two groups. Logistic regression revealed that the incidence of LC was independently affected by serum levels of myristic acid, palmitic acid, linoleic acid, eicosapentaenoic acid (EPA), lysophosphatidic acid (LPA) (18:1), glycolic acid, lysophosphatidylcholine (lysoPC) (22:6), and succinylacetone (R2=0.837, P<0.001).

Conclusions: This prospective study showed the clinical relevance of dysregulation of various metabolisms on the incidence of LC. Additionally, myristic acid, palmitic acid, linoleic acid, EPA, LPA (18:1), glycolic acid, lysoPC (22:6), and succinylacetone were identified as independent variables affecting the incidence of LC.

Disclosure

H. Yoo: None. M. Kim: None. Y. Han: None. C. Yoonjeong: None. J. Lee: None.

Funding

Korean Ministry of Health and Welfare (HI14C2686); Korean National Research Foundation (NRF-2012M3A9C4048762, NRF-2017R1C1B2007195)
286-OR: Phase 3 Results for Dasiglucagon as a Fast and Effective Treatment for Severe Hypoglycemia
Authors: PIEBER THOMAS R. , HOEVELMANN ULRIKE , WILLARD JULIE , PLUM-MOERSCHEL LEONA , MOURITZEN ULRIK , TEHRANCHI RAMIN , ARONSON RONNIE
Published: June 2019

Introduction: Severe hypoglycemia is one of the most feared adverse outcomes of insulin therapy and requires urgent assistance. The use of currently available glucagon products for rescue treatment of severe hypoglycemia is limited by complex reconstitution steps required before injection, as these products are not stable in solution. As a result, there is compelling evidence that glucagon products are under-prescribed and underused. Dasiglucagon is a stable peptide analog of human glucagon, to be administered via a ready-to-use auto-injector, in development for treatment of severe hypoglycemia.

Methods: In this pivotal Phase 3 trial, the clinical efficacy and safety of 0.6 mg dasiglucagon was compared to placebo and with reference to GlucaGen®. In total, 168 patients with T1D were randomized (2:1:1) and dosed following induced hypoglycemia to reach the glucose target of 55 mg/dL. The primary endpoint was time to PG recovery, defined as first PG increase ≥20 mg/dL after treatment initiation without rescue glucose.

Results: The primary endpoint, time to plasma glucose recovery, was found to be faster for dasiglucagon than placebo. The median time to recovery was 10 min for dasiglucagon vs. 12 and 40 min for GlucaGen® and placebo, respectively (Dasiglucagon vs. Placebo: p<0.001). In the dasiglucagon arm, 65% of patients had recovered within 10 min vs. 49% of patients receiving GlucaGen®. Similarly after 15 min, 99% of patients treated with dasiglucagon had recovered vs. 95% of patients treated with GlucaGen®. There were no serious adverse events in any groups. Nausea and vomiting were reported with similar frequencies for dasiglucagon and GlucaGen® (nausea: 55% and 53%, vomiting: 23% and 19%, respectively).

Conclusion: Dasiglucagon 0.6mg was demonstrated to be a safe, fast and effective treatment option for severe hypoglycemia.

Disclosure

T.R. Pieber: Advisory Panel; Self; ADOCIA, Arecor Limited, AstraZeneca, Novo Nordisk A/S, Sanofi. Speaker's Bureau; Self; Novo Nordisk A/S. U. Hoevelmann: None. J. Willard: None. L. Plum-Moerschel: None. U. Mouritzen: Employee; Self; Zealand Pharma A/S. R. Tehranchi: Employee; Self; Zealand Pharma A/S. R. Aronson: Research Support; Self; AstraZeneca, Becton, Dickinson and Company, Boehringer Ingelheim Pharmaceuticals, Inc., Eli Lilly and Company, Janssen Pharmaceuticals, Inc., Medtronic, Merck & Co., Inc., Novo Nordisk Inc., Sanofi, Senseonics.
288-OR: Novel Epigenetic Mechanism for Maternal Exercise Effects on Offspring Metabolic Health
Authors: KUSUYAMA JOJI , WAGNER ANA B. , HIRSHMAN MICHAEL F. , GOODYEAR LAURIE J.
Published: June 2019

Obesity and type 2 diabetes in mothers can initiate a vicious cycle of multi-generational metabolic dysfunction. Conversely, maternal exercise (MatEx) in mice improves offspring metabolic health. We previously found that the mechanism for this critical benefit of MatEx involves multiple epigenetic changes inducing increased expression of TCA and fatty acid oxidation (FAO) genes in offspring liver, leading to enhanced liver function. Here, we hypothesize that maternal high fat diet (MatHFD) and MatEx result in histone modifications, an important means of epigenetic regulation. C57BL/6 female mice were fed chow or HFD, housed with (trained) or without (sedentary) running wheels for 2 weeks before breeding and through gestation. Offspring were chow fed and sedentary and livers studied up to age 52 weeks. Trimethylation of lysine 4 on histone H3 (H3K4me3), a hallmark of active transcription, was decreased in promoters of TCA and FAO genes in offspring livers from MatHFD. Importantly, this decrease was fully reversed in offspring livers from MatEx. In fact, offspring livers from MatEx had increased H3K4me3 in these genes as early as embryonic day 13.5 and at all times after birth. Altered H3K4me3 was associated with changes in H3K4 methyltransferase (HKMT) activity, as MatHFD decreased, and MatEx increased HKMT activity in offspring livers. MatHFD induced protein carbonylation in offspring liver, and MatEx prevented this harmful effect on protein function. LC-MS/MS showed that WDR82, a regulatory component of HKMT, was robustly carbonylated in offspring liver of MatHFD, an effect reversed by MatEx. Overexpression of WDR82 in offspring hepatoblasts from MatHFD recovered H3K4me3 levels and hepatic gene expression. In sum, these data demonstrate that maternal exercise reverses the detrimental effects of maternal HFD on H3K4me3 function, likely via protection of WDR82 from carbonylation. We have identified a novel epigenetic mechanism by which maternal exercise improves offspring metabolic health.

Disclosure

J. Kusuyama: None. A.B. Wagner: None. M.F. Hirshman: Stock/Shareholder; Self; Abbott Laboratories, AbbVie Inc., Amgen Inc., Colgate-Palmolive Company, Medtronic. L.J. Goodyear: None.

Funding

National Institutes of Health (to L.J.G.); Sunstar Foundation (to J.K.)
292-LB: The Exercise-Like Impacts of Conditioned Media Harvested from Cultured Dental Pulp Stem Cells on Myokine Expressions in Skeletal Muscles
Authors: KANADA SAKI , MAKINO ERIKO , NAKAMURA NOBUHISA , MIYABE MEGUMI , ITO MIZUHO , HATA MASAKI , SAIKI TOMOKAZU , MINATO TOMOMI , MIYAZAWA KEN , GOTO SHIGEMI , MATSUBARA TATSUAKI , NARUSE KEIKO
Published: June 2019

Exercise improves not only glycemic control, but also other metabolic disorders in diabetic patients. Furthermore, exercise also stimulate muscle-derived secreted factors, myokines, via muscle contraction. The receptors of myokines expressed in the whole body and myokines affect autocrine, paracrine, and systemic functions for all people and patients with diabetes. In this study, we have demonstrated that there are exercise-like impacts of conditioned media from cultured dental pulp stem cells (DPSCs) on myokine expressions in skeletal muscles. DPSCs were isolated and cultured from the mandibular incisors of 6-week-old male Sprague-Dawley (SD) rats. Conditioned media from DPSCs (DPSC-CM) was collected after 24-hour culture of DPSCs in the serum-free medium and concentrated by a factor of 10. We firstly investigated the effects of DPSC-CM on exercise-induced myokines, follistatin-like 1 (Fstl1), Fractalkine, and fibroblast growth factor 21 (FGF-21) in C2C12 myotubes. DPSC-CM significantly increased the gene expressions of Fstl1 (209±16%, P<0.01), Fractalkine(182±41%, P<0.05), and FGF-21 (1021±39%, P<0.01) in C2C12 myotubes. To confirm these effects in vivo, we administered DPSC-CM (1.0 ml/rat) into 10 sites of unilateral hindlimb skeletal muscles of male SD rats, and examined the effects on myokine gene and protein expression. DPSC-CM significantly increased Fstl1 gene expression in the gastrocnemius muscles (477±140%, P<0.05). Immunohistochemical staining revealed that the expression of Fstl1 was increased in the DPSC-CM-injected gastrocnemius muscles. These results suggest that there is a novel exercise-like impact of DPSC-CM on skeletal muscles.

Disclosure

S. Kanada: None. E. Makino: None. N. Nakamura: None. M. Miyabe: None. M. Ito: None. M. Hata: None. T. Saiki: None. T. Minato: None. K. Miyazawa: None. S. Goto: None. T. Matsubara: None. K. Naruse: None.
293-OR: Flexible Insulin Therapy: Untethered Insulin Regimen Using Insulin Degludec and Continuous Subcutaneous Insulin Infusion (CSII) in Avidly Exercising Patients with T1D: FIT Untethered Study
Authors: ARONSON RONNIE , LI AIHUA , BROWN RUTH E. , MCGAUGH SARAH , YAVELBERG LOREN , RIDDELL MICHAEL
Published: June 2019

Athletes with T1D using CSII routinely remove their pump for prolonged periods, predisposing to hyperglycemia and ketosis. We examined the safety and efficacy of an “Untethered” approach - evenly splitting basal insulin between adjusted CSII and a daily injection of insulin degludec. This randomized cross-over study explored glycemic outcomes following in-clinic, fasted exercise (90-minute moderate- and 30-minute high-intensity). After a 1-week run-in, subjects were randomized to Usual CSII or Untethered CSII, for 5-week periods including a week of optimization, and then crossed over. Nineteen patients (age 34 ± 10 (mean ± SD), T1D duration 16 ± 11 years; mean A1C 7.1 ± 0.5) have completed both periods. In the 6 hours following moderate exercise, the Untethered arm showed improved mean glucose and spent 61% more time in range relative to the Usual arm (p<0.001) (Table). Similar trend was seen following high intensity exercise. There were no differences in pre-exercise glucose or total daily insulin dose (TDD). Both regimens were well-tolerated with no serious adverse events. An “Untethered” regimen of injected basal insulin and adjusted-rate CSII is safe and effective in avidly exercising subjects with T1D using CSII and may offer improved glycemic control in the immediate period following moderate-intensity exercise.

Disclosure

R. Aronson: Research Support; Self; AstraZeneca, Becton, Dickinson and Company, Boehringer Ingelheim Pharmaceuticals, Inc., Eli Lilly and Company, Janssen Pharmaceuticals, Inc., Medtronic, Merck & Co., Inc., Novo Nordisk Inc., Sanofi, Senseonics. A. Li: None. R.E. Brown: None. S. McGaugh: None. L. Yavelberg: None. M. Riddell: Advisory Panel; Self; Xeris Pharmaceuticals, Inc. Research Support; Self; Dexcom, Inc. Speaker's Bureau; Self; Insulet Corporation, Medtronic MiniMed, Inc. Stock/Shareholder; Self; Zucara Therapeutics Inc.

Funding

Novo Nordisk
294-OR: Type 1 Diabetes Patients with Residual Beta-Cell Function Display Improved Time in Euglycemia and Less Glycaemic Fluctuation after Exercise
Authors: TAYLOR GUY S. , SMITH KIERAN , SCRAGG JADINE , BASHIR AYAT , ORAM RICHARD A. , MCDONALD TIMOTHY J. , STEVENSON EMMA J. , SHAW JAMES A. , WEST DANIEL J.
Published: June 2019

Aims: Exercise can disrupt glycemia in individuals with type 1 diabetes (T1D). A large inter-individual variability in the glucose responses to exercise exists. It is not known how residual beta-cell function in long duration T1D influences glycaemia around exercise.

Methods: Individuals with T1D completed a postprandial urine C-peptide creatinine ratio test (UCPCR), and 25 participants (M/F 14/11, 39.4±12.4 years, HbA1c 58.8±9.3 mmol/mol-1, BMI 25.6±4.0 kg/m2, diabetes duration 21.4±13.7 years) with a range of UCPCR were recruited. Participants were split into undetectable, microsecretors and clinically significant C-peptide groups (<0.001, 0.001-0.2 and >0.2 nmol/mmol, respectively). After a maximum exercise test, participants attended the second visit whereby they completed 45 minutes of walking at 60%VO2peak. Participants then left the laboratory, with the remainder of the trial under free-living control. Blinded continuous glucose monitoring (CGM) was captured for 12 hours before exercise and for 3 days afterwards. Data (mean±SD) were analysed by one way ANOVA with significance accepted at p ≤0.05.

Results: CGM data were successfully captured for 22/25 participants. In the 12 hours prior to exercise, there was no significant differences in CGM between groups. In the first 12 hours post-exercise, the clinical C-peptide group spent significantly more time (83.7±23.2%) in time in euglycemia (3.9-10 mmol/l) compared to the microsecretors (38.8±16.9%, p =0.002) and undetectable participants (40.5±23.3%, p =0.005). The clinical group had lower post exercise SD (1.95±1.26) than the microsecretors (3.56±1.09 p =0.045) and tended to be lower than the undetectable (3.54±1.10 p =0.07).

Conclusion: Residual beta-cell function may offer protection against exercise induced dysglycemia. C-peptide could potentially be a method of stratifying the amount of exercise related support patients need to manage glucose around exercise.

Disclosure

G.S. Taylor: None. K. Smith: None. J. Scragg: None. A. Bashir: None. R.A. Oram: Other Relationship; Self; Randox Laboratories Ltd. T.J. McDonald: None. E.J. Stevenson: None. J.A. Shaw: Advisory Panel; Self; Sanofi. Other Relationship; Self; Novo Nordisk A/S. D.J. West: Research Support; Self; Arla Foods Ingredients, Dexcom, Inc.

Funding

Diabetes Research & Wellness Foundation
296-LB: Novel Analogs of the Mitochondrially Encoded Peptide CB5064 Improve Body Weight and Glucose Tolerance in Dio Mice, and Demonstrate Selective Agonism at the Apelin Receptor
Authors: GRINDSTAFF KENT , STENGER EMILY , SHANG ROBIN , YU TRACY , LUO WENDY C. , KIM VIRNA V. , PEREZ-TILVE DIEGO , CUNDY KENNETH C.
Published: June 2019

Metabolic dysregulation is integral to the development and pathogenesis of many age-related diseases, including type 2 diabetes and obesity. Several mitochondrially encoded peptides have previously been shown to be secreted from cells and to act as regulatory signals for metabolic homeostasis, including MOTS-c, SHLP2, and humanin. Novel modified analogs of a newly identified mitochondrially encoded peptide, CB5064, have been synthesized and evaluated for plasma stability, activity in a broad screen of G protein-coupled receptor (GPCR) target engagement, and for potential biological activity in diet-induced obese (DIO) mice. In vitro evaluation demonstrated that CB5064 analogs interact with and activate the apelin receptor (APJ/AGTRL1/APLNR). The observed EC50 values of several novel CB5064 analogs at the apelin receptor were in the low µM range, in some cases achieving maximum responses similar to the natural substrate, Apelin-13. In DIO mice, once daily administration of the same series of CB5064 analogs demonstrated a range of effects on metabolic regulation, including significant reduction in body weight, selective fat mass loss, and improved glucose tolerance. The apelin receptor is widely expressed in tissues and appears to play a critical role in energy metabolism, cardiovascular function, fluid homeostasis, angiogenesis, and in diabetic complications. These mitochondria-based peptide analogs therefore represent an entirely new class of molecules with agonist activity at the apelin receptor and potential for use as a source of novel therapeutics for type 2 diabetes, cardiovascular disease, and other age-related disorders. Additional evaluation of the therapeutic potential of these peptides is ongoing.

Disclosure

K. Grindstaff: Employee; Self; CohBar, Inc. E. Stenger: None. R. Shang: Other Relationship; Self; CohBar Inc. T. Yu: None. W.C. Luo: None. V.V. Kim: None. D. Perez-Tilve: Research Support; Self; CohBar, Inc., Novo Nordisk A/S. K.C. Cundy: Employee; Self; CohBar, Inc. Stock/Shareholder; Self; CohBar, Inc.
297-OR: Usefulness of Coefficient of Variation (CV) to Assess Hypoglycemia in Older Adults with Type 1 Diabetes (T1D)
Authors: TOSCHI ELENA , SLYNE CHRISTINE , DUFOUR ALYSSA B. , ATAKOV-CASTILLO ASTRID , CARL SAM , SIFRE KAYLA , GREENBERG JORDAN , MUNSHI MEDHA
Published: June 2019

Background: New metrics generated by Continuous Glucose Monitoring (CGM) have highlighted the importance of glucose variability, measured as coefficient of variation. It is also suggested that CV may be a more accurate measure to assess time spent in hypoglycemia, as opposed to A1c. This association in older adults is not well studied.

Methods: Older adults (age ≥65 years) with T1D underwent 2 weeks of CGM. Based on the international consensus, CV was calculated by dividing standard deviation of glucose by mean glucose from CGM. The participants were stratified by CV ≤ 36%, or >36%. Duration of clinically significant hypoglycemia (glucose level ≤ 54 mg/dl) and A1c were measured.

Results: We evaluated 76 older adults (mean age 71 years, duration of T1D 39 years, 62% on insulin pump, and 38% on multiple daily injections (MDI)). In participants with CV ≤36% (n=19, 25%), A1c was 6.8±0.8%, and they spent 9.6 min/day in hypoglycemia, while in those with CV>36% (n=57, 75%), A1c was 7.5±0.8%, and they spent 43 min/day in hypoglycemia (p=0.003 and p<0.0001, respectively). In participants with CV>36%, those using MDI had longer duration of hypoglycemia compared to those using insulin pump (61 minutes/day vs. 33 minutes/day, p=0.03), despite similar A1c (7.5% in both groups).

Conclusion: In our cohort of older adults with T1D, CV>36% is associated with higher duration of hypoglycemia despite higher A1c. Keeping CV value <36% in a vulnerable population may help mitigate risk of hypoglycemia independent of A1c value.

Disclosure

E. Toschi: Advisory Panel; Self; Lexicon Pharmaceuticals, Inc. C. Slyne: None. A.B. Dufour: None. A. Atakov-Castillo: None. S. Carl: None. K. Sifre: None. J. Greenberg: None. M. Munshi: Consultant; Self; Lilly Diabetes, Sanofi.

Funding

National Institute of Diabetes and Digestive and Kidney Diseases
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<5x10-8; 59 novel), which were delineated to 475 distinct signals (p<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 <1.4x10-4, Bonferroni correction). T2D-associated variants were jointly enriched (p<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 (>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 >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 <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<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<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