bluuebunny/crossref_metadata_2025 huggingface dataset:
1746-P: Differential Depletion and Expansion of Autoreactive CD8+ T Cell Subsets after Alefacept Treatment in Type 1 Diabetes (T1D)
Authors: WIEDEMAN ALICE E. , MUIR VIRGINIA S. , NEPOM GERALD T. , LINSLEY PETER , LONG S. ALICE
Published: June 2019

The T1DAL clinical trial, NCT00965458, demonstrated that alefacept (LFA-3-Ig) treatment was associated with preservation of c-peptide in subjects with new-onset T1D. Previous analysis of peripheral blood from these subjects showed reduction of CD2-expressing lymphocytes, including memory T cell populations. We now have interrogated the islet antigen-specific (autoreactive) T cell profiles from subjects in this trial using single-cell mass cytometry in combination with peptide-loaded MHC class I tetramer (Tmr) staining. We then applied a new analytical method, DISCOV-R, for phenotyping rare cell subsets using a multiparameter clustering algorithm. As expected among polyclonal CD8+ T cells, alefacept treatment preferentially reduced memory subsets, particularly those with higher expression of CD2 (Helios- transitional memory, exhausted effector memory, and Helios- effector memory CD45RA+). Prior to treatment, autoreactive Tmr+ CD8+ T cells were phenotypically heterogeneous, with three prominent phenotypes across T1D subjects (CXCR3+ naïve, Helios- and Helios+ transitional memory). Following alefacept treatment, the overall frequencies of autoreactive cells did not consistently change; however, their phenotypes were altered. CD2-high CD8+ T cells were diminished; naïve cells were expanded; and notably, a stem cell-like memory phenotype, indicative of early memory with high proliferative potential but low effector function, was also substantially expanded among autoreactive cells in most individuals. Similar phenotypic alterations occurred among polyclonal CD8+ T cells, indicating that the effects were not limited to the islet-specific T cell population. These findings suggest that alefacept treatment alters the composition of autoreactive CD8+ T cells that are not depleted, and that phenotype, not just frequency, of autoreactive cells is an important consideration for development of other immunomodulatory therapies.

Disclosure

A.E. Wiedeman: None. V.S. Muir: Research Support; Self; Celgene Corporation. G.T. Nepom: Advisory Panel; Self; GlaxoSmithKline plc., Pfizer Inc. Research Support; Self; JDRF, National Institute of Allergy and Infectious Diseases. Other Relationship; Self; Diabetes UK. P. Linsley: None. S. Long: None.
1749-P: A Case Report of IgE-Mediated Insulin Allergy Successfully Managed with a Fixed Combination of Degludec and Liraglutide
Authors: GREGORIC NADAN , OBLAK MAJA RAVNIK , KOPAC PETER
Published: June 2019

Background: Insulin allergy is a rare condition that poses a great challenge particularly in diabetics who are insulin dependent. If feasible, switching to different antidiabetic agents or immunotherapy is the common approach, but the universal consensus on insulin allergy management in diabetics is lacking. Here we present a clinical case of a diabetic with allergy to insulin successfully managed with a combination of degludec and liraglutide.

Case Report: A 51-year-old female patient with type 2 diabetes presented with poorly controlled glycaemia (HbA1c 8.6%) on dual oral antidiabetic treatment, metformin and sitagliptin. Due to pronounced fasting hyperglycaemia the NPH insulin before bedtime, but soon began to complain of urticariform skin lesions on insulin injection sites. Despite the adverse reaction the patient continued to up-titrate the insulin, but the effect on glycaemia was negligible. After switching to different NPH insulin manufacturer, to no avail, the insulin analogs detemir and glargin were introduced, respectively, but produced same adverse reaction. The elevated levels of specific IgE immunoglobulins to human insulin (2.58 klU/L) confirmed the allergy to insulin. Conversely, the novel insulin analog degludec yielded no skin reaction. The patient was then titrated to a fixed combination of degludec and GLP-1 receptor agonist liraglutide which provoked no allergic reaction and promptly improved glycemic control (HbA1c 6.7% 4 months after treatment commencement). Understandably, treatment with sitagliptin was stopped, whereas metformin was continued.

Conclusion: The patient with confirmed IgE mediated allergy to insulin exhibited allergic reaction to all types of basal insulin but degludec. It can be assumed that the unique structure of degludec has different epitopes than other insulin analogues and therefore provides safe and combined with GLP-1 receptor agonist effective therapeutic approach to patient with allergy to insulin.

Disclosure

N. Gregoric: Other Relationship; Self; Lilly Diabetes, Novartis Pharmaceuticals Corporation, Novo Nordisk A/S. M. Ravnik Oblak: None. P. Kopac: Speaker's Bureau; Self; AstraZeneca, Novartis Pharmaceuticals Corporation, Shire.
175-LB: Effect of a Telemedicine Diabetic Foot Ulcer Intervention on Health and Well-Being: A Cluster Randomized Controlled Trial
Authors: IVERSEN MARJOLEIN MEMELINK , IGLAND JANNICKE , SMITH-STRøM HILDE , OSTBYE TRULS , TELL GRETHE S. , SKEIE SVEIN , COOPER JOHN GRAHAM , GRAUE MARIT
Published: June 2019

Objective: To assess change in health and well-being using generic and disease-specific self-report instruments among patients with diabetes-related foot ulcers (DFU) receiving a telemedicine intervention compared to patients receiving standard hospital outpatient care.

Research Design and Methods: This cluster randomized controlled trial (RCT) included 182 adults with DFU (94/88 in the telemedicine/standard care groups) in 42 municipalities/districts, recruited from three clinical sites in Western Norway. The main purpose of the trial was to evaluate whether ulcer healing time in the telemedicine intervention was non-inferior to standard outpatient care. The intervention group received care in the community in collaboration with specialist health care via telemedicine; the control group received standard care. In the current study secondary endpoints as health and well-being were evaluated by four generic health and well-being questionnaires (e.g., WHO-5, EQ-5D) and two disease-specific questionnaires (Neuro-QoL, PAID). We used linear mixed-effects regression to investigate differences in change in the sum scores between the intervention and control group at the end of follow-up. Results are reported as regression coefficients with 95% CIs.

Results: In intention to treat analyses, differences between treatment groups were small and non-significant for the generic health and well-being scale scores, as well as for diabetes-related distress and foot ulcer-specific quality of life.

Conclusion: This is the first RCT assessing the effect of a telemedicine diabetic foot ulcer intervention on health and well-being. There were no significant differences in change in sum scores for health and well-being between the intervention and control group. TM was non-inferior to SOC regarding healing time. Further studies are needed to investigate long-term intervention effects on new or recurrent foot ulcers and corresponding health and well-being.

Disclosure

M.M. Iversen: None. J. Igland: None. H. Smith-Strøm: None. T. Ostbye: None. G.S. Tell: None. S. Skeie: None. J.G. Cooper: Advisory Panel; Self; AstraZeneca, Boehringer Ingelheim International GmbH. Speaker's Bureau; Self; Novo Nordisk A/S. M. Graue: None.

Funding

Norwegian Directorate of Health and Innovation; Western Norway Regional Health Authority (911716, 911605); Norwegian Diabetes Association; Western Norway University of Applied Sciences; Norwegian Research Council (221065)
1754-P: Follistatin-Like Protein 1 (FSTL1) Plays an Important Role in Human Primary Adipocyte Metabolism
Authors: AHN CHUL WOO , CHOI ARIM , KIM JUNG HYE , PARK KAHUI , LEE SANG BAE , NAM JUNGEUN , PARK SU JEONG , KIM MIN KYUNG , KIM YU SIK , NAM JI SUN , KANG SHINAE , PARK JONG SUK
Published: June 2019

Objective: FSTL1, an extracellular glycoprotein, is a novel myokine that is secreted by skeletal muscle. However, its functions in metabolism remain unclear, Adipose tissue is one of the important organs for maintaining energy homeostasis. FSTL1 is known to induce inflammatory response and inhibit insulin-mediated Akt signaling pathway, we aimed to investigate whether FSTL1 play metabolic roles in human adipose tissue. We hypothesized that FSTL1 increases lipolysis through its putative receptor DIP2A (Disco interacting protein 2 homolog A).

Methods: Human primary adipocytes were treated with recombinant FSTL1 in dose and time dependent manner. To determine its lipolytic activity, pHSL ser563,565,660, perilipin, AMPK, PKA were determined by western blot and the rate of free fatty acid was assessed. Expression of DIP2A was determined by PCR and western blot. DIP2A was knocked-down to examine whether it mediates metabolic effects of FSTL1.

Results: FSTL1 did not affect catecholamine-induced lipolysis via PKA. FSTL1, however, increased lipolysis through AMPK activation and, subsequently, HSL ser565 phosphorylation in dose dependent manner. It was found that DIP2A expression did not change during differentiation of human primary adipocyte, suggesting that it is not involved in adipocyte maturation.

Conclusion: FSTL1 increased basal lipolysis through AMPK activation. AMPK is well known sensor of the intracellular energy state and serves to regulate various signals. Its activation provides ATP by FA oxidation. Therefore, FSTL1 may play an important in the regulation of energy homeostasis. FSTL1 also has effect through DIP2A. These findings suggest that DIP2A plays important roles in FSTL1 mediated lipolysis.

Disclosure

C. Ahn: None. A. Choi: None. J. Kim: None. K. Park: None. S. Lee: None. J. Nam: None. S. Park: None. M. Kim: None. Y. Kim: None. J. Nam: None. S. Kang: None. J. Park: None.
1761-P: Brain-Restricted CB1-Receptor Antagonists Increase Oxygen Consumption Rate and Promote Beiging in 3T3-L1 Adipocytes Similar to Rimonabant Suggesting that Central Effects Can Be Avoided
Authors: PASZKIEWICZ REBECCA K. , BERGMAN RICHARD N. , FRANK AARON , WOOLCOTT ORISON O. , SOUZA SANTOS ROBERTA DE , IYER MALINI S. , CLEGG DEBORAH , KABIR MORVARID
Published: June 2019

We have previously demonstrated in dogs that following exposure to a high fat diet for 16 weeks, the brain and peripheral penetrating CB1R antagonist Rimonabant (RIM) promoted adipose tissue beiging and decreased adipocyte cell size. It is hoped that peripherally restricted CB1R antagonists, such as AM6545, will allow for the substantial beneficial metabolic effects of CB1R antagonism without the possible negative consequences associated with brain penetrance. Studies demonstrated beneficial metabolic effects of AM6545 in adipose tissue including increased energy expenditure and decreased adipocytes cell size. We thus compared the effects of RIM versus AM6545 on oxygen consumption and beiging in the well-characterized 3T3-L1 cell line model. Mature 3T3-L1s were treated for 4 or 48 hours with RIM, AM6545, vehicle or isoproterenol (ISO) (positive control). Baseline oxygen consumption and the proportional contribution of proton leak to maximal respiratory capacity was greater for cells exposed to RIM, ISO, and, in lesser extent, AM6545 after 48 hours, demonstrating greater mitochondrial uncoupling. Further, markers of adipose tissue beiging (UCP1, Prdm16, PGC1α) were also unregulated by approximately 2-fold (P<0.05) at 48 hours with RIM treatment and, in lesser extent, with AM6545. Hence, peripheral and central CB1R antagonists act directly on 3T3-L1 to increase mitochondrial function and adopt features of beige adipocytes. Thus, a direct role for CB1R antagonism on adipocytes does not require brain penetrance, supporting the importance of peripheral CB1R antagonism pharmacology for reducing the incidence of obesity and type 2 diabetes.

Disclosure

R.K. Paszkiewicz: None. R.N. Bergman: Consultant; Self; Zafgen, Inc. A. Frank: None. O.O. Woolcott: None. R. de Souza Santos: None. M.S. Iyer: None. D. Clegg: None. M. Kabir: None.
1764-P: Insulin Sensitive Induction of Adipogenesis
Authors: PEDERSEN THOMAS Å. , PEICS JULIA , OLSEN GRITH SKYTTE
Published: June 2019

Healthy adipose tissue has a remarkable ability to adapt to energy intake through contraction and expansion. A large body of evidence pin-points the ability expand adipose tissue as vital to maintain whole body lipid homeostasis and insulin sensitivity in humans as well as in mice. Increases in fat mass can be driven by expansion of existing adipocytes or by recruitment of new adipocytes from adipose tissue pre-courser cells or preadipocytes (Adipogenesis). Failure to expand adipose tissues as well as appearance of dysfunctional large adipocytes both result in local and systemic insulin resistance. Conversely, strategies to preserve adipocyte function during tissue expansion through enhanced adipogenesis could preserve insulin function in the face of obesity or high caloric intake.

In order to study the hormonal control of adipose tissue expansion we have examined the ability of insulin and IGF-1 to induce adipogenesis in isolated rodent primary SVF cells. We find that both insulin and IGF-1 dose dependently induce lipid droplet accumulation and expression of adipocyte genes in primary cells isolated from mouse epididymal and subcutaneous adipose depots. Surprisingly, adipogenesis in mouse SVF cells is a very insulin sensitive process and more specifically we identify CD45neg/Ly6c low SVF cells isolated from epididymal fat pads as extremely sensitive to insulin induced adipogenesis. These results indicate that insulin signaling directly in preadipocytes could drive adipocyte differentiation in vivo e.g., during high insulin levels in serum in response to prolonged periods of high caloric intake or lipid induced insulin resistance.

Disclosure

T.Å. Pedersen: Employee; Spouse/Partner; Cook Medical. Employee; Self; Novo Nordisk A/S. J. Peics: Employee; Self; Novo Nordisk A/S. G.S. Olsen: Employee; Self; Novo Nordisk A/S.
1765-P: Adipose Sirtuin 6 Drives Macrophage Polarization Toward M2 through IL-4 Production and Maintains Systemic Insulin Sensitivity in Mice and Human
Authors: SONG MI-YOUNG
Published: June 2019

Adipose tissue inflammation is a reproducible feature of obesity and obesity-linked insulin resistance. Here, using adipocyte-specific Sirt6 knockout mice, we explored whether adipocyte Sirt6 inhibits adipose tissue inflammation and its underlying mechanism. aS6KO mice fed normal chow gained more body weight and fat mass than wild type mice and exhibited glucose intolerance and systemic insulin resistance. Measurement of plasma and tissue cytokines and flow cytometric analysis of adipose stromal vascular cells indicated a decrease in alternatively activated M2 macrophages in adipose tissue of aS6KO mice. Mechanistically, Sirt6 regulated the expression of a canonical type 2 cytokine IL-4 by adipocytes in a cell autonomous manner, which in turn affects M2 macrophage polarization. Consistent with animal experiment data, the degree of obesity and insulin resistance demonstrated by body mass index, fasting blood glucose and HbA1c correlated negatively with the expression of Sirt6 in human visceral fat tissues. Collectively, these results suggest that adipocyte Sirt6 regulates body weight gain and insulin sensitivity independent of diet, and the increased IL-4 production by Sirt6 and resultant M2 polarization of adipose tissue macrophages may attenuate proinflammatory responses in adipose tissue.

Disclosure

M. Song: None.
1774-P: Maternal Apical Periodontitis Promotes Insulin Resistance in Adult Offspring
Authors: TSOSURA THAÍS V. , CHIBA FERNANDO Y. , MATTERA MARIA S. , PEREIRA RENATO F. , GARBIN CLÉA A. , CINTRA LUCIANO T. , SUMIDA DORIS H.
Published: June 2019

Background: Adverse stimuli applied during early fetal development can increase the risk of diseases in adulthood.

Aim: This study investigated the plasma concentrations of glucose, insulin, and tumor necrosis factor-α (TNF-α) of rats with maternal apical periodontitis (AP). We also explored the effect of maternal inflammation on the initial steps of insulin signaling and the inflammatory pathway in the gastrocnemius muscle (GM) of adult offspring.

Methodology: Fifteen female Wistar rats were distributed into control group (CN), group with 1 AP (1AP) and group with 4 AP (4AP). Thirty days following induction of AP, female rats from all groups were mated with healthy male rats. When male offspring reached 75 days of age, plasma concentrations of glucose, insulin, and TNF-α were quantified. Insulin resistance was evaluated by the homeostasis model assessment of insulin resistance (HOMA-IR) index. Phosphorylation status of pp185 tyrosine, insulin receptor substrate 1 (IRS-1) serine, IκB kinase α/β (IKKα/β), and c-Jun N-terminal kinase (JNK) in the GM were measured by western blot.

Results: Maternal AP promotes insulin resistance, impairs the initial steps of insulin signaling, increases plasma concentrations of insulin and TNF-α, and enhances IKKα/β phosphorylation in the GM of adult offspring. However, maternal AP does not seem to affect fasting glycemia and JNK phosphorylation in the GM of adult offspring.

Conclusions: Maternal AP promotes important alterations in insulin signaling and inflammation pathways, and leads to insulin resistance in adult offspring.

Disclosure

T.V. Tsosura: None. F.Y. Chiba: None. M.S. Mattera: None. R.F. Pereira: None. C.A. Garbin: None. L.T. Cintra: None. D.H. Sumida: None.

Funding

São Paulo Research Foundation (2017-01128-1)
1785-P: Mice with Muscle-Specific Insulin Receptor Overexpression Are Protected from Diet-Induced Obesity and Glucose Intolerance but Develop Post-Receptor Insulin Resistance
Authors: WANG GUOXIAO , CAI WEIKANG , BATISTA THIAGO M. , SOFTIC SAMIR , KAHN C. RONALD
Published: June 2019

Skeletal muscle insulin resistance is a prominent feature of type 2 diabetes (T2D) that precedes and predicts the development of disease in high risk populations. In individuals with T2D or pre-T2D, this insulin resistance is due, at least in part, to a decrease in the number of functional insulin receptors (IR) that localize on the cell membrane. To restore the number of IR, we generated mice that specifically overexpress IR in skeletal muscle (IRMOE). On chow diet, IRMOE mice had similar body weight compared to control, but displayed reduced body fat and higher lean mass percentage. Basal phosphorylation of IR, IRS and Akt were significantly higher in muscles from IRMOE mice compared to control, resulting in their improved glucose tolerance. Surprisingly, however, after i.v. injection of insulin, stimulated Akt phosphorylation was significantly lower in IRMOE muscle compared to control, despite persistent higher phosphorylation in IR and IRS, indicating activation of negative regulators in the IR signaling pathway when IR is chronically activated. Consistent with this, euglycemic-hyperinsulinemic clamp studies revealed significantly lower glucose infusion rate in insulin stimulated IRMOE mice compared to control. Increased basal and decreased insulin stimulated Akt phosphorylation was also observed in muscles of IRMOE mice after challenge with HFD. However, when fed HFD, IRMOE mice displayed both improved glucose tolerance and insulin tolerance which was in part due to being resistant to diet-induced obesity, despite consuming more calories. Taken together, these data indicate that muscle specific IR overexpression can protect mice from diet-induced obesity and its effects on glucose metabolism, but that chronic overexpression of IR can lead to post-receptor desensitization indicating additional post-receptor regulators of insulin signaling, which could provide new targets for therapy of T2D.

Disclosure

G. Wang: None. W. Cai: None. T.M. Batista: None. S. Softic: None. C. Kahn: Advisory Panel; Self; MedImmune. Board Member; Self; Kaleido Biosciences. Consultant; Self; AntriaBio, Inc., Cobalt Therapeutics, Flagship Pioneering. Research Support; Self; Alnylam.

Funding

American Diabetes Association (9-17-CMF-016 to G.W.)
1798-P: Insulin Modulates Dopaminergic Tone in the Human Brain—A Combined PET/MRI Study
Authors: KULLMANN STEPHANIE , BLUM DOMINIK , JAGHUTRIZ BENJAMIN , BENDER BENJAMIN , HAERING HANS-ULRICH , FOUGÈRE CHRISTIAN LA , PREISSL HUBERT , FRITSCHE ANDREAS , REIMOLD MATTHIAS , HENI MARTIN
Published: June 2019

Insulin and dopamine signaling in the human brain are known modulators of whole-body glucose metabolism. Brain insulin resistance contributes to impaired postprandial glycemia and could predispose to type 2 diabetes. Experimental evidence suggests a crucial interaction between insulin and dopamine with dopaminergic dysfunction in brain insulin resistance. We now investigated the presence of such an interaction in the human brain.

In 10 healthy young men (age 27±3 years, BMI 23.6±2.3 kg/m², HOMA-IR 1.8±1.1), we performed combined PET/MRI measurements using dynamic [11C]-Raclopride in combination with the intranasal application of 160 U insulin and placebo on two separate days in randomized, blinded order. Dopamine receptor binding potential as a measure of dopaminergic tone was estimated in regions of interests (ROI). Intranasal insulin delivery to the human brain increased dopamine receptor binding potential in the bilateral ventral and dorsal striatum, a major dopaminergic brain region (pFWE<0.001 cluster level). This response tended to be stronger in participants with higher BMI but was not statistically associated with HbA1c, fasting glucose, or HOMA-IR.

Hence, insulin delivery to the human brain influences dopaminergic tone in the striatum. As earlier studies found insulin sensitivity of this region to be involved in the postprandial modulation of whole-body insulin sensitivity, the detected mechanism might be involved in the regulation of postprandial glycaemia. Alterations in this mechanism could therefore predispose to type 2 diabetes, a hypothesis that must be tested in further studies.

Disclosure

S. Kullmann: None. D. Blum: None. B. Jaghutriz: None. B. Bender: None. H. Haering: None. C. la Fougère: None. H. Preissl: None. A. Fritsche: None. M. Reimold: None. M. Heni: Research Support; Self; Boehringer Ingelheim International GmbH, Sanofi. Speaker's Bureau; Self; Lilly Diabetes, Merck Sharp & Dohme Corp., Sanofi.

Funding

German Federal Ministry of Education and Research
1800-P: Transcriptomic Evidence of a Role for Neuron-Glia Interactions in Diabetes Remission Induced by the Central Action of Fibroblast Growth Factor 1 (FGF-1)
Authors: BENTSEN MARIE A. , RAUSCH DYLAN , MIRZADEH ZAMAN , SCARLETT JARRAD , BROWN JENNY M. , ALONGE KIMBERLY M. , MEEK THOMAS H. , SECHER ANNA , JORGENSEN RASMUS , PERS TUNE , SCHWARTZ MICHAEL W.
Published: June 2019

Lasting remission of hyperglycemia can be achieved in rodent models of type 2 diabetes by a single intracerebroventricular (icv) injection of fibroblast growth factor 1 (FGF-1), and the mediobasal hypothalamus (MBH) was recently identified as a target for this effect. To investigate the cellular basis of FGF-1 action in the MBH, we combined whole tissue RNA-sequencing with large-scale single cell and single nuclei RNA-sequencing to generate >60,000 single cell transcriptomes from diabetic ob/ob mice harvested 5d after a single icv injection of either FGF-1 or vehicle. Of the 4 known FGF receptor genes, 3 were expressed in the MBH, primarily by glial cells rather than neurons. Employing weighted gene co-expression network analysis, distinct modules that correlated with icv FGF-1 treatment were identified, again primarily in glial cells. In astrocytes, the response to FGF-1 closely paralleled the neuroprotective “A2” transcriptional phenotype induced by stroke, and both the proportion of differentiating oligodendrocytes and genes involved in extracellular matrix remodeling of the perivascular space were also upregulated by FGF-1. By comparison, the neuronal response to icv FGF-1 was one of generalized inhibition, including decreased expression of Agrp and Pmch (encoding agouti-related peptide and pro-melanin-concentrating hormone (MCH), respectively). Since Agrp and MCH neurons are activated in ob/ob mice, and since this effect predisposes to hyperglycemia, these findings support a model in which sustained inhibition of these and perhaps other neuronal subsets underlies diabetes remission induced by icv FGF-1. Moreover, this neuronal inhibition appears to be driven by neuroprotective glial responses elicited by FGF-1. These data add to growing evidence of a crucial role for neuron-glia interactions in hypothalamic control of glucose homeostasis.

Disclosure

M.A. Bentsen: None. D. Rausch: None. Z. Mirzadeh: Consultant; Self; Novo Nordisk A/S. J. Scarlett: None. J.M. Brown: None. K.M. Alonge: Research Support; Self; Novo Nordisk Inc. T.H. Meek: Employee; Self; Novo Nordisk Inc. A. Secher: Employee; Spouse/Partner; Gubra. Employee; Self; Novo Nordisk A/S. Speaker's Bureau; Spouse/Partner; Gubra. Stock/Shareholder; Self; Novo Nordisk A/S. R. Jorgensen: Employee; Self; Novo Nordisk A/S. T. Pers: None. M.W. Schwartz: Consultant; Self; Novo Nordisk A/S. Research Support; Self; Novo Nordisk A/S.

Funding

National Institutes of Health; Novo Nordisk Foundation; Lundbeck Foundation; Novo Nordisk A/S
1801-P: Common Hepatic Artery Denervation Does Not Alter the Counterregulatory Response to Hypoglycemia in an Insulin Resistant Canine Model
Authors: KRAFT GUILLAUME , MOORE L. MERKLE , FARMER BEN , SCOTT MELANIE F. , EDGERTON DALE S. , WILLIAMS PHILLIP E. , AZAMIAN BOBAK , CHERRINGTON ALAN D.
Published: June 2019

With the recent interest in sympathetic modulation as a way to improve insulin sensitivity and insulin secretion, it is important to assess the safety of the procedure. Five dogs were fed a high fat high fructose diet for 5-week before undergoing a hyperinsulinemic hypoglycemic clamp (insulin infused IV at a rate of 1.5 mU/kg/min). Then the dogs underwent a common hepatic artery surgical denervation (CHAD n=5) and were studied 1, 2 and 3 months post-surgery with the same hypoglycemic challenge. At the end of the protocol, the norepinephrine content in the liver (7±3 ng/g) and pancreas (91±34 ng/g) were significantly lower than levels in the livers and pancreas of 4 sham denervated animals (460±100 and 590±93 ng/g respectively), indicating successful denervation. There were no significant differences in fasting insulin, glucose or glucagon concentrations over the 3 months (see Table). The insulin and glucose concentrations during the last 90 min of the hypoglycemic challenge prior to and post-CHAD were also not different. The peak in plasma glucagon and epinephrine levels occurred at 30 and 120 min respectively and were similar in magnitude pre- and post-CHAD. Four sham denervated animals had the same response to the insulin induced hypoglycemic challenge as CHAD dogs.

In conclusion, the response to hypoglycemia was not affected by sympathetic denervation of the common hepatic artery.

Disclosure

G. Kraft: Consultant; Self; Metavention. L. Moore: None. B. Farmer: None. M.F. Scott: None. D.S. Edgerton: Speaker's Bureau; Self; Novo Nordisk A/S. P.E. Williams: None. B. Azamian: Employee; Self; Metavention. A.D. Cherrington: Board Member; Self; Biocon, Fractyl Laboratories, Inc., Metavention, Sensulin, LLC., vTv Therapeutics, Zafgen, Inc. Consultant; Self; Abvance, Boston Scientific Corporation, California Institute for Biomedical Research, Galvani Bioelectronics Limited, MedImmune, Novo Nordisk Inc., Thetis Pharmaceuticals LLC. Research Support; Self; Abvance, Diasome Pharmaceuticals, Inc., Novo Nordisk Inc. Stock/Shareholder; Self; Abvance, Biocon, Fractyl Laboratories, Inc., Metavention, Novo Nordisk Inc., Sensulin, LLC., Thetis Pharmaceuticals LLC, Zafgen, Inc.

Funding

Metavention, Inc.
1802-P: Nr1d1/Nr1d2 in POMC Neurons Regulates Circadian Eating Behaviors and Energy Expenditure
Authors: ZHOU WENJUN , YANG TINGTING , LI XIN , SUN ZHENG
Published: June 2019

Proopiomelanocortin (POMC) neurons in the arcuate nucleus of ventral hypothalamus and the nucleus tractus solitarius are involved in many biological functions. Nuclear receptor Nr1d1 and Nr1d2 are the key component of the molecular circadian clock machinery and are widely expressed in the brain. Here we address the role of Nr1d1/Nr1d2 in POMC neurons using conditional knockout mouse models. Mice with Nr1d1 and Nr1d2 specifically depleted in POMC neurons (KO) displayed a diet-dependent metabolic dysregulation. When fed with normal chow, KO mice show similar body weight, food intake, and energy expenditure as control WT mice. However, KO mice are resistant to high-fat diet (HFD)- induced obesity even though they consume slightly more food. Energy expenditure and the temporal pattern of food intake was altered in the KO mice fed with HFD when compared to control. These findings revealed the pivotal role of Nr1d1 and Nr1d2 of the POMC neurons in regulating the eating behavior and energy expenditure.

Disclosure

W. Zhou: None. T. Yang: None. X. Li: None. Z. Sun: None.

Funding

American Diabetes Association (1-19-PDF-012 to W.Z.)
1810-P: Effect of HM15211, a Novel Long-Acting GLP-1/GIP/Glucagon Triple Agonist in the Neurodegenerative Disease Models
Authors: WONKI KIM , KIM JEONG A. , LEE SANG HYUN , BAE SUNGMIN , CHOI IN YOUNG , KIM YOUNG HOON
Published: June 2019

Metabolic disturbances such as diabetes and obesity, are a potential risk factors for progressive neurodegenerative disease such as Parkinson’s disease (PD), Alzheimer’s disease (AD), and multiple sclerosis (MS). There is evidence that an antidiabetic or obesity drug of the glucagon-likepeptide-1(GLP-1) family has direct neuroprotective effects in an experimental models of neurodegenerative disease. Recently, we have developed HM15211, a novel long-acting GLP-1/GIP/Glucagon triple agonist. Previous studies have shown that HM15211 with a unique activity profile exerted neuroprotective effects in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) induced subchronic PD mice model. In the present study, we evaluated 1) the neuroprotective effects of HM15211 in chronic MPTP/probenecid PD model, 2) the protection of AD progression in diabetic model, and 3) neuroprotective effect in the relapsing-remitting experimental autoimmune encephalomyelitis (EAE) model of MS. In chronic PD mice model, HM15211 administration protected dopaminergic neuronal death and decreased alpha synuclein in striatum. HM15211 also significantly improved the MPTP/probenecid induced motor impairments in behavior tests. In addition, HM15211 increased phosphorylated tau, and reversed inflammatory cytokines and oxidative stress marker in aged db/db mice, which have a pathological characters of AD. These results suggest that HM15211 have a protective effect in progression from diabetes to AD. In another experiment, HM15211 have shown the anti-inflammatory and neuroprotective effects in the EAE mouse model. HM15211 significantly reduced the EAE clinical score and resulted in the reversal of histopathological sequelae compared to vehicle. Furthermore, HM15211 decreased pro-inflammatory cytokines, while it upregulated anti-inflammatory cytokines. Based on these observations, HM15211 might be a potential therapeutic option for the neurodegenerative diseases.

Disclosure

K. Wonki: Employee; Self; Hanmi Pharm. Co., Ltd. J.A. Kim: Employee; Self; Hanmi Pharm. Co., Ltd. S. Lee: Employee; Self; Hanmi Pharm. Co., Ltd. S. Bae: None. I. Choi: Employee; Self; Hanmi Pharm. Co., Ltd. Y. Kim: None.
1814-P: Type 2 Diabetes Increases Stroke Severity through Sustained Hyperperfusion of Cerebral Vasculature following Ischemic Stroke in Rats
Authors: RAMSAY CIARAN DAVID , HOWELLS DAVID W. , KESKE MICHELLE , SUTHERLAND BRAD , PREMILOVAC DINO
Published: June 2019

Stroke is a leading cause of death and permanent disability. Type 2 diabetes (T2D) increases the incidence of stroke by four-fold and is a strong predictor of worse outcome following stroke with greater disability and mortality than in nondiabetics. While the reasons for increased stroke severity in T2D are unclear, we hypothesized that T2D alters the cerebral vascular response to ischemic stroke leading to reduced vascular reperfusion and hence greater infarct following restoration of blood flow post-stroke. To address this hypothesis, 15-week-old male Sprague Dawley rats were given a high fat diet and treated with osmotic-mini pump infused STZ (110mg/kg over 14 days) to develop a mild T2D phenotype, characterized by obesity and glucose of 8-10mM. Normal chow-fed aged matched rats were used as controls. Four weeks after induction of T2D, we quantified cerebral perfusion in real-time using contrast enhanced ultrasound (CEU) before, during and immediately after middle cerebral artery occlusion, the gold standard model of ischemic stroke. Following this, rats were allowed to recover and a battery of behavioral tests was used to assess stroke severity over 14 days. A final CEU measure was obtained 14 days post-stroke to track vascular changes over time. Cerebral perfusion tended to be increased in T2D rats at baseline but was similarly reduced during middle cerebral artery occlusion. Strikingly, T2D rats had a 3-fold increase in cerebral perfusion immediately post-stroke and this hyperperfusion was sustained 14 days post-stroke. Importantly, although there was no difference in behavior at baseline or immediately following stroke, by day 6, T2D rats developed a greater level of disability compared with control rats that was sustained until day 14.

In conclusion, we demonstrate that T2D alters the cerebral vascular response to ischemic stroke, leading to severe vascular hyperperfusion and that this is associated with a greater level of disability in rats.

Disclosure

C.D. Ramsay: None. D.W. Howells: None. M. Keske: None. B. Sutherland: None. D. Premilovac: None.
182-LB: Circulating Adhesion Molecules and Relationships to Micro and Macrovascular Outcomes: The TODAY Study
Authors: TRYGGESTAD JEANIE B. , SHAH RACHANA , BRAFFETT BARBARA , BACHA FIDA , GIDDING SAMUEL , GUBITOSI-KLUG ROSE , SHAH AMY SANGHAVI , URBINA ELAINE M. , KATZ LORRAINE E.
Published: June 2019

Atherosclerosis begins in childhood, accelerating in those with obesity, hypertension and type 2 diabetes (T2DM). Binding of mononuclear cells to vascular endothelium, initiated by cellular adhesion molecules and chemokines, is an early step in this process. The TODAY study, a clinical trial of adolescents with T2DM, included three treatment arms. The primary end-point was treatment failure defined as A1c persistently >=8% over 6 months period or metabolic decompensation. The aims of the present analysis are to evaluate: 1) changes in ICAM, VCAM, E-Selectin and MCP-1 over time 2) whether treatment group influences levels of these biomarkers and 3) the association of these markers with macrovascular and microvascular outcomes. Participants from the TODAY cohort (n=515) with at least two of the four annual assessments of adhesion molecules at baseline, 12, 24, 36 months were included. Over the study period, there were significant increases in ICAM (p=0.002) and MCP-1 (p<0.001) and significant decreases in VCAM (p<0.001) and E-Selectin (p=0.016) across the entire cohort. There were no significant treatment group interactions. Participants who reached the primary outcome had higher E-Selectin compared to participants with sustained control over the duration of the study (p<0.0001). ICAM increased 2.0% (p<0.001), VCAM increased 1.5% (p<0.001), E-selectin increased 7.0% (p<0.001), and MCP-1 increased 1.4% (p=0.001) for every 1% increase in HbA1c. E-selectin was 9.5% (p<0.001) and 5.2% (p=0.017) higher and ICAM was 5.9% (p=0.001) and 4.5% (p=0.046) higher in participants with microalbuminuria or hyperfiltration respectively. E-selectin increased with increasing left ventricular mass and TAPSE, a measure of right ventricular function, (p=0.043 and p=0.001, respectively). This analysis demonstrates cellular adhesion molecules increase with increasing HbA1c in youth with T2DM, and ICAM and E-selectin are associated with worsening micro and macrovascular outcomes.

Disclosure

J.B. Tryggestad: None. R. Shah: None. B. Braffett: None. F. Bacha: Research Support; Self; National Institute of Diabetes and Digestive and Kidney Diseases, Pediatric Diabetes Consortium. Other Relationship; Self; AstraZeneca, Jaeb Center for Health Research. S. Gidding: Research Support; Self; Color Genomics. R. Gubitosi-Klug: None. E.M. Urbina: None. L.E. Katz: None.

Funding

National Institute of Diabetes and Digestive and Kidney Diseases
1829-P: Acute Inhibition of Intestinal Neprilysin Enhances Glucose-Stimulated Insulin Secretion in Mice
Authors: ESSER NATHALIE , MUNDINGER THOMAS O. , BARROW BREANNE , MONGOVIN STEVE M. , ZRAIKA SAKENEH
Published: June 2019

Increased activity of the ubiquitous peptidase neprilysin in obesity and type 2 diabetes (T2D) is associated with impaired glucose homeostasis, and its inhibition has been shown to improve β-cell function. Neprilysin cleaves and inactivates insulinotropic peptides, including some produced in the intestine (e.g., GLP-1, GIP, CCK). While neprilysin is expressed in the intestine, the contribution of intestinal neprilysin to modulate β-cell function remains unknown. Thus, we sought to determine whether acute selective pharmacological inhibition of intestinal neprilysin enhances glucose-stimulated insulin secretion (GSIS) in healthy mice and could be a therapeutic target in T2D. High (20 mg/kg) or low (0.05 mg/kg) dose neprilysin inhibitor thiorphan, or vehicle, was orally administered to chow-fed C57BL/6 mice, then plasma and tissues were collected after 40 minutes to determine neprilysin activity. High dose thiorphan significantly inhibited neprilysin activity in plasma and all tissues listed below (data not shown). In contrast, low dose thiorphan inhibited neprilysin activity by 91±5% in colon, 50±12% in ileum and 62±4% in jejunum vs. vehicle (n=7-8, p<0.05), without altering neprilysin activity in pancreas, kidney and plasma. Thus, low dose thiorphan was used to selectively inhibit intestinal neprilysin in a separate cohort of mice 30 minutes prior to an oral glucose tolerance test (OGTT; 2g/kg glucose) in which GSIS was determined. Compared to vehicle, thiorphan significantly increased insulin levels during the OGTT (incremental AUC 0-30 minutes: 3181±606 vs. 5786±600 pmol/L.minutes-1, n=6-7, p=0.01) without altering basal insulin (95±21 vs. 114±25 pM, n=6-7, p=0.6).

In summary, under physiological conditions, acute selective inhibition of intestinal neprilysin increases GSIS in vivo. Thus, intestinal neprilysin plays a role in modulating β-cell function, and strategies to inhibit neprilysin specifically in the intestine might improve β-cell dysfunction in T2D.

Disclosure

N. Esser: None. T.O. Mundinger: None. B. Barrow: None. S.M. Mongovin: None. S. Zraika: Research Support; Self; Novartis Pharmaceuticals Corporation.

Funding

National Institutes of Health (DK098506); Belgian American Educational Foundation; Société Francophone du Diabète
1831-P: Delayed Glucagon Suppression Contributes to Postprandial Hyperglycemia Independently of the Degree of Insulin Secretion
Authors: ADAMS J.D. , LAURENTI MARCELLO C. , RIZZA ROBERT A. , EGAN AOIFE M. , COBELLI CLAUDIO , MAN CHIARA DALLA , VELLA ADRIAN
Published: June 2019

Type 2 diabetes (T2DM) is characterized by impaired postprandial insulin secretion and glucagon suppression. Recent work suggests that α-cell dysfunction is independent of β-cell dysfunction. The effect of diabetes-associated variation in TCF7L2 on α-cell function also suggests a direct role in the pathogenesis of T2DM. To quantify the interaction between a lack of glucagon suppression with intact, mildly or moderately impaired insulin secretion we studied 45 nondiabetic subjects with and without suppressed glucagon. On both occasions, an intravenous glucose infusion mimicking the systemic appearance of 50g oral glucose was administered in the presence of a pancreatic clamp. Glucagon was infused at a rate of 0.65ng/kg/min, starting at 0 min or 120 min (non-suppressed vs. suppressed respectively). In one group (100%) a nondiabetic insulin profile was infused; another group received 80% of this profile and the third group received 60% of the insulin infused in the first group. We observed that a lack of glucagon suppression increased peak (p < 0.05) and area above basal (AAB) glucose (p < 0.02) in all groups (1.5 ± 0.1 vs. 1.1 ± 0.1; 1.6 ± 0.1 vs. 1.1 ± 0.1; 1.5 ± 0.1 vs.1.3 ± 0.1 Mol per 5hr, 100%, 80%, 60% respectively). These increases were explained by delayed suppression of endogenous glucose production (EGP) so that EGP at 120 minutes was higher (p < 0.05) in the presence of non-suppressed glucagon (6.6 ± 1.0 vs. 4.2 ± 0.6; 7.2 ± 0.8 vs. 5.1 ± 0.5; 7.7 ± 0.8 vs. 5.9 ± 0.6 μmol/kg/min, 100%, 80%, 60% respectively) as was integrated EGP (p < 0.05) over the postprandial period (-1.9 ± 0.4 vs. -2.1 ± 0.1; -1.8 ± 0.1 vs. -1.9 ± 0.2; -1.6 ± 0.3 vs. -1.8 ± 0.1 mmol/kg per 5hr, 100%, 80%, 60% respectively). These data indicate that impaired postprandial glucagon suppression contributes to postprandial hyperglycemia through increased EGP even when insulin secretion is intact as can occur early in prediabetes. These effects are exacerbated by increasing degrees of insulin deficiency.

Disclosure

J. Adams: None. M.C. Laurenti: Employee; Self; GlySens Incorporated. R.A. Rizza: Advisory Panel; Self; Lilly Diabetes, Metavention, Novo Nordisk A/S, vTv Therapeutics. A.M. Egan: None. C. Cobelli: None. C. Dalla Man: None. A. Vella: Advisory Panel; Self; vTv Therapeutics. Research Support; Self; Novo Nordisk Inc.

Funding

National Institutes of Health
1832-P: Impact of Habitual Daily Activity on Glucose Tolerance (GT) and ß-Cell Function in Adults with Impaired GT (IGT) or Type 2 Diabetes (T2D)
Authors: TEMPLE KARLA A. , TJADEN ASHLEY H. , ATKINSON KAREN M. , BARENGOLTS ELENA , HANNON TAMARA S. , MATHER KIEREN J. , UTZSCHNEIDER KRISTINA , EHRMANN DAVID A. , MOKHLESI BABAK ,   RISE CONSORTIUM
Published: June 2019

We examined the relationship between habitual daily activity and measures of GT, insulin sensitivity and β-cell responses in 230 adults (mean age 55 y; BMI 35 kg/m2) with IGT or drug-naïve, recently diagnosed T2D. Participants underwent a 3-h OGTT and hyperglycemic clamp. Wrist accelerometers worn for 7 days measured total activity counts (TAC; daily mean 233,461 {∼ 50th %ile for age}). We evaluated whether TAC predicted fasting plasma glucose (FPG); OGTT 2-h plasma glucose (2-h G) or glucose incremental area under the curve (G-iAUC); hyperglycemic clamp measures of insulin sensitivity (steady-state glucose infusion rate/insulin [M/I]) and β-cell responses (acute C-peptide response to glucose [ACPRg], steady-state C-peptide, maximal β-cell response [ACPRmax]); and OGTT C-peptide index (CPI, [ΔC-Peptide0-30/ΔGlucose0-30]). There was no association of TAC with FPG, 2-h G, or G-iAUC. Higher TAC was associated with higher insulin sensitivity. After adjusting for M/I, higher TAC was associated with lower CPI, but not with clamp-based β-cell responses. Despite similar post-load glucose values, adults with IGT or T2D who were more active demonstrated a greater than expected reduction in C-peptide response given their higher insulin sensitivity. Further studies are needed to understand this dissociation of activity effects on insulin sensitivity and β-cell function.

Disclosure

K.A. Temple: None. A.H. Tjaden: None. K.M. Atkinson: None. E. Barengolts: None. T.S. Hannon: Advisory Panel; Self; Eli Lilly and Company. K.J. Mather: Advisory Panel; Self; Roche Diabetes Care. Research Support; Self; Abbott, Merck & Co., Inc., Novo Nordisk A/S, Sanofi. K. Utzschneider: Consultant; Self; Novo Nordisk Inc. D.A. Ehrmann: None. B. Mokhlesi: None. R. Consortium: None.

Funding

National Heart, Lung, and Blood Institute; National Institute of Diabetes and Digestive and Kidney Diseases
1833-P: PERK Inhibitors Enhanced Glucose-Stimulated Insulin Secretion in a Mouse Model of Type 2 Diabetes
Authors: YANG YE SEUL , KIM MIN JOO , KIM MI NA , KIM JIWON , YOON KUN-HO , PARK KYONG SOO , JUNG HYE SEUNG
Published: June 2019

Background: PERK (pancreatic endoplasmic reticulum (ER) kinase) has been reported to have important implications in the generation, function, and viability of β cells. We have reported that partial PERK attenuation using PERK inhibitors (PI) enhanced glucose-stimulated insulin secretion (GSIS) from pancreatic islets, through induction of ER chaperone BiP and regulation of ER calcium, but not through the EIF2A-ATF4 pathway. In the current study, we investigated if PI have the same effects under metabolic stress condition.

Methods: GSK2606414, a specific PERK inhibitor, was treated to mouse islets under 20-mM glucose and 0.5-mM palmitate to examine GSIS. Male C57BL/6J mice were fed with high-fat diet combined with streptozotocin injections to mimic type 2 diabetes (DM). Several doses (6 ∼ 16 mg/kg/day) of GSK2656157 and glimepiride were administrated to the mice for 8 weeks, and metabolic phenotypes were evaluated such as body weight, blood glucose levels, insulin secretion and sensitivity, and then changes in the pancreas were measured.

Results: High-glucose and palmitate treatment significantly increased PERK phosphorylation in the isolated islets. Suppression of GSIS and glucose-stimulated Ca2+ transit was also observed. PI at 40 nM which decreased PERK phosphorylation by 40% significantly recovered the GSIS and cytosolic calcium. In the mice where significant weight gain, insulin insufficiency, and prominent hyperglycemia were induced, PI at 10 mg/kg/day significantly enhanced GSIS and reduced blood glucose levels compared to the vehicle. The effects were similar to those by 10 mg/kg/day of glimepiride. Administration of PI did not induce changes in beta cell mass or pancreatic insulin contents, however, high dose PI decreased pancreatic weight.

Conclusion: PI at low dose significantly enhanced GSIS in vitro and in vivo under metabolic stress, and improved hyperglycemia in the mice mimicking type 2 DM, suggesting a potential as a new therapeutic approach for type 2 DM.

Disclosure

Y. Yang: None. M. Kim: None. M. Kim: None. J. Kim: None. K. Yoon: Advisory Panel; Self; AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Merck Sharp & Dohme Corp., Novo Nordisk Inc. Speaker's Bureau; Self; Takeda Pharmaceutical Company Limited. K. Park: None. H. Jung: None.

Funding

Korean Ministry of Health and Welfare (A062260)
1835-P: Deletion of Insulin Receptor Substrate 2 in AGRP Neurons Causes Beta-Cell Dysfunction
Authors: TAO RONGYA , COPPS KYLE D. , WHITE MORRIS F. , STOEHR OLIVER
Published: June 2019

In humans, type 2 diabetes progresses when beta-cells fail to compensate peripheral insulin resistance. Insulin receptor substrate (IRS) proteins mediate a variety of the metabolic and growth-promoting actions of insulin and IGF-1, and are important integrators of heterologous regulation. Systemic ablation of the Irs2 gene in mice causes β-cell dysfunction that prevents compensatory insulin secretion while insulin resistance progresses to life threatening hyperglycemia. Upregulation of IRS2 in pancreatic β-cell using rat insulin-2 promoter (Irs2RIP) promotes sufficient hyperinsulinemia to prevent diabetes. Targeted Irs2 deletion using CreRIP also causes beta-cell failure and hyperglycemia, but the mechanism is unclear because CreRIP drives deletion in beta-cells and hypothalamus—especially in the arcuate nucleus (ARC). AGRP neurons in the ARC play an important role in regulating energy homeostasis, but it is unknown whether IRS2 in the AGRP neurons might also affect beta-cell function. Here we show that disruption of Irs2 (but not Irs1) in AGRP neurons (Irs2f/fCreAgrp-mice) reduced islet mass by 45%. To test insulin secretory function, we subjected mice to hyperglycemic clamps. The glucose infusion rate to maintain steady-state hyperglycemia decreased significantly. Moreover, insulin secretion failed to respond to L-arginine treatment in aged Irs2f/fCreAgrp-mice. Moreover, Irs2f/fCreAgrp-mice displayed progressive deterioration of glucose homeostasis and a lack of beta -cell compensation during high fat diet. Overall, our results indicate that dysfunction of Irs2 in AGRP neurons might contribute to the pathophysiology of type 2 diabetes.

Disclosure

R. Tao: None. K.D. Copps: None. M.F. White: Advisory Panel; Self; Housey Pharmaceutical Research Laboratories. O. Stoehr: None.

Funding

National Institutes of Health (5R01DK098655-06)