bluuebunny/crossref_metadata_2025 huggingface dataset:
Contrast Enhanced Ultrasound with Submicron-Sized Contrast Agents Detects Diabetes Progression in Mouse Models of Type 1 Diabetes
Authors: RAMIREZ DAVID , BENNINGER RICHARD K.
Published: June 2018

In type 1 diabetes (T1D) the decline in beta-cell mass occurs many years prior to clinical presentation. A non-invasive method to diagnose insulitis and beta-cell mass decline prior to diabetes does not exist. During T1D progression, the islet microvasculature increases permeability due to insulitis. Iron oxide nanoparticle MRI contrast accumulation has previously been suggested as a measure of ongoing insulitis. Contrast enhanced ultrasound (CEUS) using gas-filled microbubbles (MBs) measures acoustic backscatter at sub-harmonic frequencies and is clinically approved. Sub-micron sized ‘nanobubbles’ (NBs) have previously been developed and show extravasation in tumors. Here we tested whether these sub-micron sized NBs show disease-mediated extravasation in mouse models of T1D. We performed CEUS measurements following NB infusion in NOD mice and control (NOD;Rag1ko and C57Bl6) mice. We also examined fluorescent coverage (FC) following rhodamine-labeled bubble delivery in these animals, through isolating pancreata for histology and quantifying FC and insulitis scores. We observed progressive increased CEUS signal accumulation in pancreas of 4w and 10w NOD mice, which was absent in NOD kidneys (p < 0.001). Rag1ko controls showed minimal CEUS signal in pancreas and kidney. The FC was limited to the pancreata of NOD mice and was substantially higher in endocrine compared to exocrine tissue (p = 0.002). NOD mouse islets also had substantially more FC compared to Rag1ko (p < 0.0001), and showed a correlation between NOD insulitis score and FC (p=0.043). Thus, CEUS with NB contrast agents can detect disease progression prior to T1D onset, which will be important for early T1D diagnosis and monitoring of disease prevention or reversal.

Disclosure

D. Ramirez: None. R.K. Benninger: None.
Glucocorticoid Receptor Is Instrumental for the Mechanism of Beta-Cell Compensation for Insulin Resistance in Dietary Obesity
Authors: YAMAUCHI JUN , LEE SOJIN , DONG HENRY
Published: June 2018

The glucocorticoid receptor (GR) is known as a receptor of steroids - potent anti-inflammatory ligands that are commonly used for the medication of allergy or asthma in children and adults. However, chronic use of steroids is associated with a significant risk of developing visceral adiposity and insulin resistance, a medical condition that is known as the Cushing syndrome. We hypothesized that excessive GR activation is deleterious to beta-cells such that genetic GR depletion would preserve beta-cell mass and function in response to metabolic stress. To address this hypothesis, we generated beta-cell specific GR knockout (KO) mice, using an inducible Pdx1-CreERT2-loxP system. We then fed beta-cell GR-KO and WT littermates both regular chow and high-fat diet for up to 30 weeks, followed by determination of beta-cell mass and function. We found that GR-deficient mice, as opposed to WT littermates, developed high blood glucose levels and glucose intolerance. This effect was not secondary to alterations in weight gain or insulin sensitivity. Instead, we detected a significant reduction in glucose-stimulated insulin secretion, correlating with abnormal glucose metabolism in GR-KO mice. We reproduced these findings in both male and female GR-KO mice. Anti-insulin immunohistochemistry revealed that GR-KO mice had significantly reduced beta-cell mass, and decreased beta-cell replication, as visualized by in vivo BrdU-labeling assay. GR deficiency appears to impair the ability of beta-cells to compensate for fat-elicited obesity and insulin resistance, resulting in prediabetes in GR-KO mice. These results unveiled an unexpected role of the glucocorticoid-GR signaling in safeguarding beta-cell mass and function in response to metabolic stress.

Disclosure

J. Yamauchi: None. S. Lee: None. H. Dong: None.
Foxo1-Expressing Cells in the Gut as a Source of Insulin for Diabetes Treatment
Authors: MCKIMPSON WENDY , KUO TAIYI , ACCILI DOMENICO
Published: June 2018

The hallmark of type 1 diabetes is the irreversible destruction of insulin-secreting β-cells in the pancreas. Consequently, an attractive therapeutic approach is to trigger the regeneration of functional β-cells elsewhere within a diseased individual. We have previously demonstrated that removal of Foxo1 in Neurogenin3 (Ngn3)-expressing cells creates functional insulin-secreting cells within the intestine. The cells, however, only convert into insulin+ cells at a low frequency, limiting their therapeutic potential. Using a novel knock-in reporter mouse that encodes a Foxo1-Venus fusion protein (FoxV), we have identified a novel population of Foxo1-expressing cells in the glandular epithelium of the stomach. These cells are distinct from other gut FoxV+ cells in that, unlike Foxo1+ cells in the intestine, stomach FoxV+ cells rarely express serotonin. To identify the nature of Foxo1+ stomach cells, we next isolated single cells from stomach and, using flow cytometry and qPCR, analyzed the gene expression of FoxV+ cells. Although Foxo1-positive cells do not express markers of chief, pit, or neck cells, they have increased transcript levels of the parietal cell marker H+/K+ ATPase. We further validated these results by immunofluorescence and FACS. To determine if elimination of Foxo1 can convert stomach cells to insulin-producing cells, we next ablated Foxo1 in either Ngn3+ or parietal cells in mice using cre-mediated recombination. Strikingly, we detected cells positive for insulin and C-peptide in the stomach of mice harboring Foxo1 deletion. Similarly, when we inactivated Foxo1 in primary stomach cultures, we detected markedly increased “β-like” gene expression and insulin+, C-peptide+ cells. Taken together we have identified a new population of Foxo1-expressing cells in the stomach and demonstrated that Foxo1 ablation is sufficient to convert these cells to insulin-producing cells.

Disclosure

W. McKimpson: None. T. Kuo: None. D. Accili: None.
Molecular Mechanism of Androgen Receptor Stimulation of Insulin Secretion in Male ß Cells
Authors: XU WEIWEI , ASHFORD FIONA B. , HODSON DAVID , MAUVAIS-JARVIS FRANCK
Published: June 2018

Androgen deprivation therapy increases the risk of type 2 diabetes mellitus (T2DM) in men. We previously showed that male pancreatic β-cell specific androgen receptor knockout (βARKOMIP) mice develop glucose intolerance because AR potentiates glucose-stimulated insulin secretion (GSIS) through increasing cyclic AMP (cAMP) production and amplifies the insulinotropic effect of glucagon-like peptide-1 (GLP-1). We show that male βARKOMIP mice exhibit impaired intraperitoneal (IP) glucose tolerance- because of impaired IP-GSIS- without alteration in oral glucose tolerance, suggesting that AR amplifies the islet-derived, but not the gut-derived GLP-1 to potentiate GSIS. Using male insulin-secreting β-cell line 832/3 transduced with exchange factor directly activated by a cAMP (EPAC)-based fluorescence resonance energy transfer (FRET) sensor, we observe that AR agonist dihydrotestosterone (DHT) allows GLP-1, glucagon and gastric inhibitory polypeptide (GIP) to increase cAMP production above level of the individual hormones and to a similar level to forskolin control. Accordingly, DHT increases the insulinotropic effect of forskolin, GLP-1, glucagon and GIP in mouse islets. The insulinotropic effect of DHT is abolished using EPAC and PKA inhibitors as well as rapamycin indicating that DHT amplifies the effect of GPCRs coupled to adenylate cyclase and stimulates GSIS via a cAMP/PKA/EPAC pathway and activation of mTOR.

Disclosure

W. Xu: None. F.B. Ashford: None. D. Hodson: None. F. Mauvais-Jarvis: Research Support; Self; Boehringer Ingelheim Pharmaceuticals, Inc..
Beta-Cell Glucotoxicity in KATP-Induced Diabetes Is Alleviated by Genetic Reduction of Glucose Metabolism
Authors: FORTUNATO MANUELA , YAN ZIHAN , SHYR ZEENAT A. , CONWAY HANNAH E. , REMEDI MARIA S.
Published: June 2018

Glucotoxicity, reduced β-cell function and mass are key events in the pathogenesis and progression of diabetes. We previously generated an insulin secretory-deficient mouse model of human KATP-induced diabetes. As predicted, KATP-GOF (gain-of-function) mice demonstrate severe diabetes due to low circulating insulin levels, reiterating the human disease. However, as diabetes progresses, KATP-GOF mice show reduction of insulin content and β-cell mass, consequences of glucotoxicity. Islets from diabetic mice also show increased glucose metabolism (NAD(P)H autofluorescence) and oxygen consumption rate (Seahorse analysis), and augmented mitochondrial membrane potential. Strikingly, they also demonstrate increased glucokinase (GK, first enzyme in glucose metabolism) and GLUT2 gene transcripts and proteins. We hypothesized that augmented glucose metabolism in diabetic mice will contribute to the glucotoxic reduction in in insulin content, β-cell function and mass; and that these consequences will be alleviated by reducing glucose metabolism. To test this in vivo, we crossed KATP-GOF mice with mildly glucose-intolerant heterozygous GK+/- knockout mice (with reduced glucose metabolism), to generate double-transgenic GK+/-/KATP-GOF mice. Progression of diabetes, assessed by blood glucose levels overtime, is slower in GK+/-/KATP-GOF mice, compared with KATP-GOF mice. Importantly also, glucose tolerance is enhanced in GK+/-/KATP-GOF mice 12 days after disease onset, with respect to KATP-GOF mice, without significant changes in insulin sensitivity. Insulin gene expression, insulin content and β-cell mass are partially preserved in GK+/-/KATP-GOF mice. Together, these results demonstrate that hyperglycemia induce accelerated metabolism in KATP-GOF mice damaging the islets, and that deceleration of metabolism by reducing GK activity delays/prevent hypermetabolism induced-glucotoxicity in KATP-GOF mice.

Disclosure

M. Fortunato: None. Z. Yan: None. Z.A. Shyr: None. H.E. Conway: None. M.S. Remedi: None.
Functional Roles of Proline/Arginine-Rich End Leucine-Rich Repeat Protein (PRELP) in Adipocytes
Authors: EGUCHI JUN , WADA JUN
Published: June 2018

Adipose tissue fibrosis is recognized as a hallmark of adipose tissue dysfunction. We have previously identified PRELP as a novel regulator of adipogenesis by using lentiviral short hairpin RNA libraries targeting murine genomes. PRELP belongs to the family of leucine-rich repeat proteins, which are characterized by a series of adjacent leucine-rich motifs flanked by disulfide-bounded domains. Functionally, PRELP anchors basement membranes to connective tissues. However, roles of PRELP in adipocytes remain unclear. In the present study, we show that PRELP expression is up-regulated both in adipocytes of high-fat diet induced obesity mice and ob/ob mice. PRELP reduction in 3T3-L1 adipocytes leads to enhanced insulin signaling and insulin-stimulated glucose uptake. To determine functions of PRELP in the development of metabolic phenotypes, we generated adipocyte-specific PRELP transgenic mice. Adipose-specific overexpression of PRELP results in the development of significant adipose tissue fibrosis and insulin resistance on high fat diet without difference of body weight and adiposity. This phenotype is associated with increased expression of inflammation genes and decreased insulin signaling in adipose tissue of PRELP transgenic mice. Furthermore, we find that global Prelp deficient mice are protected from adipose tissue fibrosis and insulin resistance associated with high fat feeding. These data suggest that PRELP might play a key role in the development of adipose tissue remodeling in obesity.

Disclosure

J. Eguchi: None. J. Wada: None.
Deletion of 12/15-Lipoxygenase Preserves Retinal Neuronal Function in Diabetic Retinopathy
Authors: WAN MING , IBRAHIM AHMED , EL-SHAFEY MOHAMED , ELMASRY KHALED , ABDULMONEIM SAMER , SAUL ALAN , AL-SHABRAWEY MOHAMED
Published: June 2018

Purpose: Diabetic retinopathy (DR) is a neurovascular complication of diabetes with critical limitations in current treatment due to a lack of neuroprotection. These limitations mandate the necessity for new therapy that considers the neuronal component of DR. The goal of this study is to explore the role of 12/15-lipoxygenase (LO) in diabetes-induced retinal neuronal dysfunction.

Methods: 12/15-LO deficient (12/15-LO−/−) mice were crossbred with Ins2Akita/+ Akita) mice, a classical model of DR, to generate a new mouse model with a double knockout (12/15-LO−/−/Akita). The generated mice were subjected to comprehensive electroretinogram (ERG) analyses after 6 months of diabetes.

Results: Profound effects of knocking 12/15-LO out on diabetes-induced reduction in visual system’s functionality were observed. First, Scotopic Threshold Response (pSTR) peak heights, which measure the functionality of ganglion cells (GCs), are consistently lower in Akita than in wild type (WT) mice. Knocking 12/15-LO out significantly improved the overall pSTR and significantly restored the loss of the GC marker (Brn3) as compared with Akita mice. Second, diabetes reduced the functionality of cone-bipolar cells by 32%, as assessed by photopic B-Wave (pBW) amplitudes, however, this reduction was significantly improved in 12/15-LO−/−/Akita mice compared with Akita mice (p<0.05, n=9). Lastly, diabetes caused a significant reduction (22%) in cone photoreceptor functionality as indicated by a lower response to natural noise stimuli (NNS) in Akita compared to WT mice (p<0.05, n=8) and this reduction was completely recovered in 12/15-LO−/−/Akita mice (p<0.05, n=9).

Conclusion: To our knowledge, this is the first report that elimination of 12/15-LO protects against diabetes-induced visual system dysfunction. The findings may lead to new therapeutic approaches in prevention and treatment of DR.

Disclosure

M. Wan: None. A. Ibrahim: None. M. El-Shafey: None. K. Elmasry: None. S. Abdulmoneim: None. A. Saul: None. M. Al-Shabrawey: None.
Adipocyte mTORC1 Suppresses Treg Cell Development and Browning of White Adipose Tissue through CRTC2/COX-2/PGs Pathway
Authors: ZHANG XING , LUO YAN , DING XIAOFENG , WANG CHUNQING , YANG XUEXIAN , LIU MEILIAN
Published: June 2018

Adipose tissue (AT) contains abundant CD4+Foxp3+ regulatory T (Treg) cells; however, the physiological signals that control AT Treg cell and the role of AT Treg cell in thermogenesis have not been defined. Here we show that obesity activated mTORC1 signaling while decreased frequency of Treg cells in white adipose tissue (WAT), and adipose-specific inhibition of mTORC1 by raptor deficiency promoted development of Treg cells and beige adipocytes and protected mice against diet-induced reduction of Treg cells and beige adipocytes in WAT and diet-induced obesity. Mechanistically, mTORC1 signaling phosphorylated CREB regulated transcription coactivator 2 (CRTC2) and subsequently suppressed expression of cyclooxygenase 2 (COX-2) and secretion of prostaglandins (PGs) including PGI2, PGD2 and PGE2 in vivo and in adipocytes. Treatment of PGE2 but not PGI2 and PGD2 promoted differentiation and activation of Treg cells in vitro and in vivo. Pharmacological inhibition of COX-2 or blocking Treg cell by neutralization of CD25 attenuated mTORC1 inhibition-induced browning of white fat. Further, deficiency of Treg cell suppressed the inducing effect of PGE2 administration on thermogenic gene expression and oxygen consumption in inguinal and epididymal WAT. Taken together, these results uncover that adipocyte mTORC1 is a key regulator of Treg cell and beige adipocyte development in adipose tissue via CRTC2/COX-2/PGs pathway-dependent mechanisms.

Disclosure

X. Zhang: None. Y. Luo: None. X. Ding: None. C. Wang: None. X. Yang: None. M. Liu: None.
Maternal Obesity, Diabetes during Pregnancy, Gestational Weight Gain, Breastfeeding, and Child’s BMI Growth Trajectory from Ages 2 to 6 Years
Authors: WANG XINHUI , MARTINEZ MAYRA P. , XIANG ANNY
Published: June 2018

Limited studies have evaluated the interplay among maternal obesity, maternal diabetes including preexisting type 1 (T1D), type 2 (T2D) or gestational diabetes mellitus (GDM), excessive gestational weight gain (EGWG), and breast feeding (BF) in association with childhood growth trajectory. Here we assessed their associations with child’s BMI trajectory from ages 2 to 6 years. Data included 71,892 singleton children born at 28-44 gestational weeks in 2007-2011 at Kaiser Permanente Southern California hospitals who had at least 5 BMI measurements between ages 2 to 6. First, BMI trajectory patterns for these children were identified using group-based trajectory modeling method. Second, the relationships between maternal exposures and growth trajectory groups were evaluated using multinomial logistic regression adjusting for maternal age at delivery, race/ethnicity, education, and child’s sex. Three distinct BMI trajectory groups were identified: group 1 (59% of the cohort) had stable low BMI over time, group 2 (35% of the cohort) had stable median BMI over time, and group 3 (6% of the cohort) had high and increasing BMI over time. Relative to group 1, the adjusted odds ratio (OR[95% CI]) of being in group 3 associated with maternal exposures were 8.0 [7.3-8.8] for pre-pregnancy obesity, 2.8 [2.6-3.1] for pre-pregnancy overweight, 2.1 [1.2-3.8] for T1D, 1.7 [1.5-2.0] for T2D, 1.2 [1.1-1.4] for GDM, 1.5 [1.4-1.6] for EGWG, and 1.3 [1.2-1.4] for BF<6 months after adjusting for each other. ORs of being in group 2 were much smaller and were insignificant for T1D and GDM. Further adjustment for birth weight and gestational age at delivery had a small effect on the OR estimates but did not change the conclusion. Thus, among the four exposures, high and increasing BMI trajectory in offspring from age 2 to 6 years was strongly associated with maternal obesity and overweight; modestly with maternal T1D, T2D, and EGWG; and slightly with BF<6 months and GDM.

Disclosure

X. Wang: None. M.P. Martinez: None. A. Xiang: None.
The Adipose-Derived Insulin Sensitizer Adiponectin Can Activate Intracellular Signaling Independent of the Well Known AdipoR1 or AdipoR2 Receptors
Authors: GUNNET JOSEPH W. , LI WENYU , WANG YUANPING , CHAVEZ JOSE A. , HUSOVSKY MATT , MENG RONG , LANG WENSHENG , JEYASEELAN JEY R. , CALDWELL GARY W. , EDAVETTAL SUZANNE
Published: June 2018

Adiponectin (ADN) is a crucial mediator of insulin sensitivity and vascular health. The discoveries of the adiponectin receptors AdipoR1 and AdipoR2 (R1 and R2) prompted efforts to identify ADN-mimetics. R1 and R2 are especially difficult drug targets due to the complex higher-order structures of ADN as well as challenges in creating receptor knock-out cell lines. We approached R1 and R2 as possible targets beginning with a rigorous validation of the ligand and receptors. ADN preparations from multiple internal and external sources were characterized in physicochemical, cell binding and in vitro assays (pAMPK/pACC and ceramidase). It is important to note that ADN from only one source passed this evaluation. This ADN triggered in vitro activities associated with this protein, including stimulation of phosphorylation of ACC (EC50 = 9.3 ug/ml), calcium influx (EC50 = 2.2 ug/ml), glucose uptake, fatty acid oxidation and ceramidase activity. With a biologically active ADN preparation identified, we strove to understand the interactions of ADN with R1 and R2. However, both R1 and R2 are ubiquitously expressed, complicating the in vitro confirmation of receptor-dependent functional activity. We selected HAP1 cells, an ADN-responsive haploid CML derived line well-suited for gene deletion via CRISPR-Cas9 technology, to generate R1, R2 and R1/R2 deficient cells. Surprisingly, the stimulatory activity of adiponectin in pACC and ceramidase assays in HAP1 cells deficient in R1, R2 or both R1 and R2 were indistinguishable from those in wild type HAP1 cells. In addition, transient overexpression of R1 in HAP1 cells or pretreatment with antibodies to block R1 did not alter their pACC responses to ADN. These data indicate R1 and R2 are not responsible for all the in vitro biological activity attributed to ADN and suggest that there must be other yet unidentified receptors for ADN.

Disclosure

J.W. Gunnet: Employee; Self; Janssen Research & Development. W. Li: None. Y. Wang: None. J.A. Chavez: None. M. Husovsky: None. R. Meng: None. W. Lang: None. J.R. Jeyaseelan: Employee; Self; Janssen Pharmaceuticals, Inc.. G.W. Caldwell: None. S. Edavettal: Employee; Self; Janssen Research & Development.
Fully Closed-Loop Glucose Control in Noncritical Care Settings—A Randomised, Controlled Two-Centre Study
Authors: BALLY LIA , THABIT HOOD , HARTNELL SARA , ANDEREGGEN EVELINE , RUAN YUE , WILINSKA MALGORZATA E. , EVANS MARK , WERTLI MARIA M. , COLL ANTHONY P. , STETTLER CHRISTOPH , HOVORKA ROMAN
Published: June 2018

Automated fully closed-loop (CL) insulin delivery system without meal-bolusing was evaluated in medical and surgical non-critical care wards of two acute hospitals in Switzerland and United Kingdom. In a randomised controlled parallel design study, 136 adults with inpatient hyperglycaemia requiring subcutaneous (s/c) insulin were randomised to receive either CL-directed s/c insulin delivery (n=70) or conventional s/c insulin therapy as per local practice with masked continuous glucose monitoring (n=66) for up to 15-days. Participants consumed self-selected hospital meals and were matched for age (68±10 vs. 67±13 years; CL vs. control), HbA1c (7.8±2.5 vs. 8.0±1.9%) and BMI (32.7±8.2 vs. 32.3±8.1kg/m2). During CL, participant’s usual insulin and sulphonylurea therapy were withheld. In an intention to treat analysis, the proportion of time when sensor glucose was in target range from 5.6 to 10.0mmol/l was significantly higher during CL (p<0.001, Table). CL reduced time spent above target (p<0.001), and reduced mean and standard deviation of sensor glucose (p<0.001). Time spent hypoglycaemic was low and comparable. Total daily insulin delivery was not different. No severe hypoglycaemia or serious adverse events occurred in either group.

In conclusion, fully closed-loop without meal-bolusing is safe, and may improve glucose control in a heterogeneous inpatient population.Closed-loop insulin delivery (n=70)Conventional insulin therapy (n=66)PTime spent at sensor glucose levels (%)5.6 to 10.0 mmol/l65.7±16.841.5±16.9<0.001>10.0 mmol/l23.7±16.649.5±22.8<0.001<5.6 mmol/l10.6±6.79.0±13.30.37<3.0 mmol/l0.0 (0.0, 0.1)0.0 (0.0, 0.0)0.80Mean sensor glucose (mmol/l)8.5±1.610.5±2.4<0.001SD of sensor glucose (mmol/l)2.6±1.03.2±1.1<0.001Total daily insulin (U)44.0 (27.5, 73.3)40.2 (26.5, 65.6)0.36Data are mean ± SD or median (IQR).

Disclosure

L. Bally: None. H. Thabit: None. S. Hartnell: Speaker’s Bureau; Self; Medtronic, Roche Pharma. E. Andereggen: None. Y. Ruan: None. M.E. Wilinska: None. M. Evans: Advisory Panel; Self; Novo Nordisk A/S, Eli Lilly and Company, Cellnovo, Roche Pharma. Speaker’s Bureau; Self; Abbott, Novo Nordisk A/S. M.M. Wertli: None. A.P. Coll: None. C. Stettler: None. R. Hovorka: Speaker’s Bureau; Self; Novo Nordisk A/S. Advisory Panel; Self; Novo Nordisk A/S. Speaker’s Bureau; Self; Eli Lilly and Company, AstraZeneca. Other Relationship; Self; B. Braun Medical Inc.. Research Support; Self; Medtronic. Other Relationship; Self; Medtronic. Research Support; Self; Abbott, JDRF, Diabetes UK, National Institute of Diabetes and Digestive and Kidney Diseases.
Human Regular U-500 Insulin via Continuous Subcutaneous Insulin Infusion vs. Multiple Daily Injections in Adults with T2D—The VIVID Study—All Randomized Population
Authors: GRUNBERGER GEORGE , BHARGAVA ANUJ , LY TRANG T. , ZISSER HOWARD , ILAG LIZA , MALONE JAMES K. , ZHANG SHUYU , JOHNSON JENNAL
Published: June 2018

VIVID is the first prospective randomized controlled trial evaluating CSII (using an investigational Omnipod U-500™ Insulin Management System) vs. MDI (TID) with U-500R (Humulin® R U-500). This was a 26-week, open-label, multicenter, parallel study in adults with T2D on high dose insulin (201-600 units/day) with or without other antihyperglycemic agents (NCT02561078). We present results for the all randomized population (N=420); results for the primary population (excluding participants on concomitant GLP-1 RAs or SGLT2 inhibitors) are reported separately. Baseline characteristics were similar between arms. Without adjusting for multiplicity, CSII demonstrated significantly greater reduction in A1C and FPG at a lower total daily dose vs. MDI at 26 weeks. Also, a higher percentage of participants on CSII reached A1C targets <7.0%, <7.5%, and <8.0%. Severe hypoglycemia rates were low. Other hypoglycemia rates were similar (documented symptomatic) or higher (nocturnal) for CSII vs. MDI. Weight gain was similar (Table). In the all randomized population, U-500R via CSII demonstrated greater efficacy at a lower dose with a higher nocturnal hypoglycemia rate vs. MDI. CSII with U-500R may be a viable option for patients with T2D on high dose insulin. Individualized dose titration will be important for safely achieving glycemic targets.

Disclosure

G. Grunberger: Research Support; Self; Medtronic MiniMed, Inc., Novo Nordisk Inc.. Speaker’s Bureau; Self; Eli Lilly and Company, Novo Nordisk Inc., Sanofi, Boehringer Ingelheim Pharmaceuticals, Inc., AstraZeneca, Janssen Pharmaceuticals, Inc.. A. Bhargava: None. T.T. Ly: Employee; Self; Insulet Corporation. H. Zisser: Employee; Self; Verily Life Sciences LLC.. Stock/Shareholder; Self; Verily Life Sciences LLC. L. Ilag: Employee; Self; Eli Lilly and Company. Stock/Shareholder; Self; Eli Lilly and Company. J.K. Malone: Employee; Self; Eli Lilly and Company. S. Zhang: Employee; Self; Eli Lilly and Company. Stock/Shareholder; Spouse/Partner; Eli Lilly and Company. J. Johnson: Employee; Self; Eli Lilly and Company.
Loss of CEACAM1 in Endothelial Cells Contributes to the Development of Cardiac Fibrosis
Authors: MUTURI HARRISON T. , GHADIEH HILDA E. , KHUDER SAJA S. , JASH SUKANTA , ABU HELAL RAGHD , DANIELS NIGEL A. , LIU JAMES , PURI VISHWAJEET , VAZQUEZ GUILLERMO , GUPTA RAJESH , NAJJAR SONIA M.
Published: June 2018

Carcinoembryonic Antigen-related Cell Adhesion Molecule 1 (CEACAM1) has been implicated in various physiological processes in the vascular systems, including blood vessel formation, and endothelial barrier function. Consistent with a role for hepatic CEACAM1 in promoting insulin sensitivity by mediating insulin clearance and maintaining physiologic plasma insulin levels, we have recently identified a role for hepatic CEACAM1 in preventing cardiac hypertrophy and dysfunction. The aim of this study was to elucidate the contribution of endothelial cell CEACAM1 in cardiac function and fibrosis that has remained elusive. To this end, we generated an endothelial cell-specific Ceacam1 knockout mouse line (VECadCc1−/−). Gomori Trichrome Blue Staining revealed a significant perivascular and interstitial fibrosis in the heart of VECadCc1−/− mice compared to their control littermates. Consistently, the mRNA levels of markers of fibrosis [collagen 1A1, α-SMA and fibroblast-specific protein1 (FSP1)] were elevated with a reciprocal decrease of endothelial cell makers (CD31), indicating endothelial mesenchymal transition (EndMT) in these mice. Cardiac fibrosis was associated with increased activation of the TGF-β1-Smad2/3 signaling pathway. Immunofluorescence staining co-localized CD31 with α-SMA in the myocardial perivascular section of VECadCc1−/− hearts. Moreover, mRNA level of α-SMA, FSP1, SNAIL and SLUG were up-regulated while that of CD31 was down-regulated in mouse heart endothelial cells (MHEC) isolated from VECadCc1−/− mice. The Akt/eNOS pathway was basally inactivated, leading to lower nitric oxide (NO) level in MHEC isolated from VECadCc1−/− mice. Moreover, loss of endothelial CEACAM1 caused reduced association with SHP-2 phosphatase and consequently, reduction in insulin-stimulated IRS-1/Akt/eNOS activation.These studies identified an important role for the loss of CEACAM1 in endothelial cells as a mechanism underlying cardiac fibrosis.

Disclosure

H.T. Muturi: None. H.E. Ghadieh: None. S.S. Khuder: None. S. Jash: None. R. Abu Helal: None. N.A. Daniels: None. J. Liu: None. V. Puri: None. G. Vazquez: None. R. Gupta: None. S.M. Najjar: None.
Hepatic Overexpression of Retinoic Acid Receptor ß Downregulates Mitochondrial Pyruvate Carrier 1 and Protects from Metabolic Dysregulation in ob/ob Mice
Authors: KEYHANI NEJAD FARNAZ , LEE DONGHWAN , CHEN HANQING , LIANG ZERONG , MUSI NICOLAS , ZANG MENGWEI
Published: June 2018

Our recent studies identify that retinoic acid receptor β (RARβ), a vitamin A-related nuclear receptor, regulates mitochondrial fatty acid oxidation in hepatocytes and in vivo in a fasting-inducible manner, which contributes to the metabolic adaptation to fasting. Mitochondrial pyruvate carrier (MPC) has been recently identified to control gluconeogenesis in hepatocytes, which contributes to the pathogenesis of diabetes. The present study aims to determine whether RARβ is linked to the control of glucose metabolism. We observed hepatic downregulation of RARβ occurred in high fat, high sucrose (HFHS) diet-fed mice and in genetically obese ob/ob mice. Treatment with all-trans-retinoic acid (RA), a natural ligand of RARβ and an approved drug for anticancer treatment, at doses of 2.5- 5.0 mg/kg/day for 4 weeks lowered fasting glucose levels and ameliorated glucose intolerance and insulin resistance in HFHS-fed mice, which was associated with increased hepatic gluconeogenic gene expression. Moreover, adenovirus-mediated overexpression of RARβ restored hepatic impairment of RARβ and systemic glucose intolerance and decreased hepatic gluconeogenesis by obesity. Mechanistically, overexpression of RARβ decreased obesity-induced expression of MPC1 and MPC2, key regulators of gluconeogenesis from pyruvate. Increasing RARβ in ob/ob mice also inhibited the induction of alanine transaminase (ALT), a key enzyme that converts alanine into pyruvate for glucose production. Moreover, overexpression of RARβ downregulated lipogenic gene expression, upregulated fatty acid oxidation, and improved hepatic steatosis and hyperlipidemia in ob/ob mice. Hepatic and systemic insulin resistance were also reduced by overexpressing RARβ. In sum, hepatic defective RARβ may contribute to the upregulation of MPC/ALT and gluconeogenesis in diabetes and RARβ can be a novel drug target to normalize metabolic dysregulation in diabetes.

Disclosure

F. Keyhani Nejad: None. D. Lee: None. H. Chen: None. Z. Liang: None. N. Musi: None. M. Zang: None.
Cognitive Function and Brain Plasticity in Obese Patients—The Impact of Bariatric Surgery
Authors: DARDANO ANGELA , DANIELE GIUSEPPE , LUNGHI CLAUDIA , CICCARONE ANNAMARIA , SANTINI FERRUCCIO , CECCARINI GIOVANNI , MORETTO CARLO , PENNO GIUSEPPE , MICCOLI ROBERTO , MORRONE MARIA CONCETTA , DEL PRATO STEFANO
Published: June 2018

Background and Aim: Obesity adversely affects brain function, but the effect of bariatric surgery on cognitive function is still poorly evaluated. We assessed metabolic parameters and cognitive function in obese patients before and after bariatric surgery (RYGB).

Methods: A 75g OGTT was performed in 10 nondiabetic obese patients (BMI 46.0±5.4 kg/m2; age 43.6±10.5 years; HbA1c 43.3±4.6 mmol/mol) before and 6-months after RYGB for measurements of metabolic parameters. Mini Mental State Examination (MMSE) and Montreal Cognitive Assessment (MoCA) were administered; neuronal plasticity (NP) of the visual cortex was measured as change in ocular dominance after 120 min monocular deprivation. No subject with drugs or alcohol abuse, psychiatric illness, head injury, brain tumor, dementia, epilepsy, learning disorder, developmental disability, and impaired sensory function were included.

Results: At baseline median MMSE score was 28.5 (range 25-30) and median MoCA score 26.5 (range 21-30), indicating early cognitive abnormalities (MoCA<26 points). Six months after RYGB, BMI decreased to 35.2±6.1 kg/m2 (p=0.004) and glucose tolerance did not change. HOMA-IR (from 5.1±2.3 to 1.6±1.1) and disposition index (from 3.3±4.6 to 35.0±41.0) all improved (p<0.05); post-OGTT GLP-1 increased from 5336.6±2262.6 to 11132.3±3412.2 pmol/lx120min (p<0.05) and fasting plasma leptin decreased from 80.0±46.7 to 13.1±6.6 pmol/l (p<0.008). NP increased from 0.03±0.1 to 0.11±0.14 (p<0.01). Cognitive performance improved in all subjects (MMSE: 30, range 28-30; MoCA: 28.5, range 24-30; both p=0.03 or less vs. baseline). There was no correlation between MoCA and metabolic changes though it was positively correlated with NP post-RYGB (p=0.04).

Conclusion: Morbid obese subjects have subclinical cognitive impairments and brain plasticity alterations that significantly improve after RYGB. While no correlation was found between metabolic and hormonal changes, NP was positively associated with MoCA score.

Disclosure

A. Dardano: None. G. Daniele: None. C. Lunghi: None. A. Ciccarone: None. F. Santini: None. G. Ceccarini: None. C. Moretto: None. G. Penno: None. R. Miccoli: None. M. Morrone: None. S. Del Prato: Advisory Panel; Self; AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Eli Lilly and Company, GlaxoSmithKline plc., Intarcia Therapeutics, Inc., Merck & Co., Inc., Novartis Pharmaceuticals Corporation, Novo Nordisk A/S, Servier, Sanofi, Takeda Pharmaceuticals U.S.A., Inc.. Research Support; Self; Merck & Co., Inc., Novartis Pharmaceuticals Corporation, Boehringer Ingelheim Pharmaceuticals, Inc., AstraZeneca. Speaker's Bureau; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Novartis Pharmaceuticals Corporation, Takeda Pharmaceuticals U.S.A., Inc.. Advisory Panel; Self; Janssen Biotech, Inc., Abbott.
Glucagon-Like Peptide-1 (GLP-1) Increases Skeletal and Cardiac Muscle, Microvascular Perfusion, and Improves Metabolic Insulin Action in Obese Humans
Authors: WANG NASUI , KIT TAN ALVIN WAI , JAHN LINDA , HARTLINE LEE , AYLOR KEVIN W. , BARRETT EUGENE , LIU ZHENQI
Published: June 2018

Obesity is associated with metabolic and microvascular insulin resistance. The latter is characterized by impaired insulin-mediated microvascular recruitment. GLP-1 engenders vasodilation in addition to its glycemic actions. At physiological concentrations, GLP-1 acutely recruits skeletal and cardiac muscle microvasculature in healthy humans and this action is preserved in the insulin resistant rodents.

To examine whether GLP-1 recruits microvasculature and improves insulin’s action in obese humans, 15 obese adults (age 27 ± 2.2, BMI 33.7 ± 1.2) were studied thrice at random order after an overnight fast. Subjects received an intravenous infusion of GLP-1 (1.2 pmol/kg/min) for 150 mins or an intravenous infusion of either normal saline or GLP-1 (1.2 pmol/kg/min) for 150 mins with a euglycemic insulin clamp (1 mU/kg/min) superimposed over the last 120 mins. Skeletal and cardiac muscle microvascular blood volume (MBV), flow velocity (MFV) and blood flow (MBF) were determined at 0, 30 and 150 min. Steady-state glucose infusion rate (GIR) was calculated.

Insulin infusion alone did not change MBV or MBF in either skeletal or cardiac muscle, confirming the presence of microvascular insulin resistance. GLP-1 infusion alone increased MBV by ∼30% and ∼40% in skeletal and cardiac muscle respectively (p<0.05) without affecting MFV, leading to a significant increase in MBF in both skeletal and cardiac muscle. Superimposition of insulin infusion to GLP-1 infusion did not further increase MBV or MBF in either skeletal or cardiac muscle, but raised the steady-state GIR from 4.1± 0.4 to 5.0 ± 0.5 mg/kg/min (p<0.05).

We conclude that in obese humans with microvascular insulin resistance GLP-1's vasodilatory actions are preserved in both skeletal and cardiac muscle microvasculature. This may contribute to improving metabolic responses to insulin and decreasing cardiovascular morbidity and mortality associated with obesity and diabetes.

Disclosure

N. Wang: None. A. Tan: None. L. Jahn: None. L. Hartline: None. K.W. Aylor: None. E. Barrett: None. Z. Liu: None.
Functional and Developmental Heterogeneity in Human Adipose Tissue Depots
Authors: JOHNSON JAMES W. , CERO CHERYL , O'MARA ALANA , LINDERMAN JOYCE D. , BASKIN ALISON S. , CYPESS AARON
Published: June 2018

Obesity results from the accumulation of excess white adipose tissue, and its increasing rates worldwide pose a significant risk to overall health due to its multiple comorbidities, like type 2 diabetes. A novel treatment strategy is to increase energy expenditure through activation of brown and beige adipocytes, which can use uncoupling protein 1 (UCP1) for thermogenesis. In addition to having different developmental lineages, brown and beige adipocytes differ functionally regarding the additional mechanisms by which they generate heat. Most human fat depots are not easily accessible, so little is known about their developmental and functional characteristics. In this study, we collected adipose tissue from 11 patients (age 16-84, 4 female/7 male) who had undergone autopsy. We sampled from 7 anatomically distinct fat depots: subcutaneous (sc), omental (om), retroperitoneal (rp), pericardial (pc), periadrenal (pa), paraspinal (ps), and supraclavicular (sv). We measured mRNA levels of lineage and functional genes associated with brown, beige, and white adipocytes. Energy storing white adipocytes were defined by high leptin expression, while energy expending brown and beige cells were defined by high UCP1; these were further distinguished as having a brown lineage using Zic1 and beige via Tbx1. Principal component analysis showed three different clusters of genes consistent with the brown, beige, and white lineages. Functionally, based on expression of leptin and UCP1 for white and brown fat, respectively, the sc and om were white (P<0.05), while rp, pc, pa, ps, and sv were mostly brown (P<0.05). Regarding lineage of the UCP1+ depots, ps had both beige and brown cells, while rp, pa, ps, and sv were predominantly brown.

In summary, human adipose tissue is not merely white or brown but instead is composed of many depots with distinct developmental lineages and functional capacities. These differences will be important when trying to use brown and beige adipocyte thermogenesis to treat obesity and diabetes.

Disclosure

J.W. Johnson: None. C. Cero: None. A. O'Mara: None. J.D. Linderman: None. A.S. Baskin: None. A. Cypess: None.
A Study of APOC3 Gene Polymorphism in Patients of Diabetes Mellitus with Hypertriglyceridemia
Authors: KALITA BIPUL C. , KOTOKEY RATAN K. , DAS SUBHALAKSHMI , CHAKRAVORTY AVISEK , SONI HARISH
Published: June 2018

Aims and Objectives: To find out the presence of Sst I polymorphism in APOC3 gene in patients of diabetes mellitus with Hypertriglyceridemia.

To find out the correlation between serum triglyceride levels and Sst I polymorphism.

To find out the correlation between serum levels of various lipoprotein fractions and Sst I polymorphism.

Methods: The present study is a hospital-based observational case-control study, carried out in 300 patients of diabetes mellitus. Of these 300 subjects, 150 were hypertriglyceridemic and served as cases, while the other 150 who were normotriglyceridemic served as controls. PCR for APOC3 gene was done in all subjects.

Result: In a total of 300 subjects, 26.33% (n=79) had shown presence of Sst I polymorphism in APOC3 gene. Polymorphic gene was present in 35.33% of cases while only in 17.33% of controls. The mean cholesterol level in the group with Sst I polymorphism was significantly higher than in group with S1 allele only. The mean triglyceride in the group with S1 allele only was 152.97±45.86 mg/dl and that in the group with Sst I polymorphism was 212.84±103.07 mg/dl, and this increased mean triglyceride level was statistically significant as determined by chi-square test. Mean HDL level was also significantly lower in the group with Sst I polymorphism as compared to that with S1 allele only. The odds ratio of developing hypertriglyceridemia was 2.6059 in those carrying the Sst I polymorphism as compared to those with only S1 allele. The association between increasing levels of triglyceride and the increasing percentage of Sst I polymorphism was also statistically significant.

Conclusion: This is the first study from North-East India to determine the Sst I polymorphism in APOC3 gene, and demonstrate a clear association between the polymorphism and serum levels of various lipoprotein fractions, including triglycerides. A prospective study comprising of a wide population group will help us in forming a definite conclusion.

Disclosure

B.C. Kalita: None. R.K. Kotokey: None. S. Das: None. A. Chakravorty: None. H. Soni: None.
Sustained Diabetes Remission Induced by the Central Action of Fibroblast Growth Factor-1 (FGF-1) Requires Activation of Integrin Receptor aνß3
Authors: SCARLETT JARRAD , BROWN JENNY M. , PHAN BAO ANH N. , KHAV EDDIE L. , MATSEN MILES E. , ACHARYA NIKHIL K. , NGUYEN HONG T. , SECHER ANNA , JORGENSEN RASMUS , MORTON GREGORY J. , SCHWARTZ MICHAEL W.
Published: June 2018

The brain is implicated in the antidiabetic effects of members of the fibroblast growth factor (FGF) peptide family. Previously, we reported that a single intracerebroventricular (icv) injection of FGF-1 induces remission of hyperglycemia that is sustained for weeks in rodent models of type 2 diabetes (T2D). The integrin receptor ανβ3 plays a key role in FGF-1 signaling, and crosstalk between ανβ3 and the FGF-1-FGF receptor (FGF-R) complex is necessary for increased DNA synthesis and proliferation in cultured cells. These cellular responses downstream of FGF-1-FGFR-ανβ3 result from sustained (e.g., lasting for 5 hours) activation of the extracellular signal-regulated kinase (ERK) signal transduction pathway. In cultured fibroblasts, prolonged ERK activation induced by FGF-1 is not replicated with a mutant FGF-1 peptide (R50E) that activates FGFR normally, but does not activate ανβ3 signaling. Similarly, we report that whereas icv FGF-1 (3 µg) induces phosphorylation of ERK1/2 at both early (20 minute) and late (5 hr) time points in hypothalamic punches obtained from wild type mice, icv injection of the same dose of R50E elicited this effect only at the early time point. Thus, R50E fails to activate hypothalamic FGF-1-FGFR-ανβ3 signaling in vivo. To investigate whether activation of ανβ3 is required for the prolonged antidiabetic effect of icv FGF-1, diabetic ob/ob mice received a single injection of either vehicle (Veh), recombinant FGF-1 (3 µg), or R50E (3 µg). Whereas R50E mimicked the effect of native FGF-1 to reduce food intake, body weight and blood glucose for the first week after icv injection (P<0.001 vs. Veh), the mutant peptide failed to elicit the sustained diabetes remission observed in animals receiving icv FGF-1. We conclude that signaling by the full FGF-1-FGFR-ανβ3 complex is necessary for the effect of FGF-1 signaling in the brain to elicit sustained remission of diabetic hyperglycemia in rodent models of T2D.

Disclosure

J. Scarlett: None. J.M. Brown: None. B.N. Phan: None. E.L. Khav: None. M.E. Matsen: None. N.K. Acharya: None. H.T. Nguyen: None. A. Secher: Employee; Self; Novo Nordisk A/S. Stock/Shareholder; Self; Novo Nordisk A/S. Employee; Spouse/Partner; Gubra. R. Jorgensen: Employee; Self; Novo Nordisk Inc.. G.J. Morton: None. M.W. Schwartz: Consultant; Self; Novo Nordisk A/S. Research Support; Self; Novo Nordisk A/S.
A Single-Cell Transcriptomics Roadmap to Investigate Diabetes Remission Induced by the Central Action of Fibroblast Growth Factor-1 (FGF-1)
Authors: BENTSEN MARIE A. , RAUSCH DYLAN , SCARLETT JARRAD , ALONGE KIMBERLY M. , TIMSHEL PASCAL N. , MIRZADEH ZAMAN , SECHER ANNA , JORGENSEN RASMUS , PERS TUNE , SCHWARTZ MICHAEL W.
Published: June 2018

Recently, lasting remission of hyperglycemia was achieved in rodent models of type 2 diabetes (T2D) by a single intracerebroventricular (icv) injection of FGF-1. While the mechanism underlying this effect is unknown, the lack of association with changes of body fat mass or risk of hypoglycaemia raises the possibility that icv FGF-1 normalizes the defended level of glycemia, rather than simply lowering blood glucose levels. Recent work from our lab has identified gluco-regulatory neurocircuits in the mediobasal hypothalamus (MBH) as targets for this FGF-1 effect. To investigate the mechanism underlying FGF-1 action in this brain area, we used large-scale single cell RNA-sequencing to identify and characterize FGF-1-responsive cells in mouse MBH. Based on >70,000 single cell MBH transcriptomes from diabetic ob/ob mice harvested 5d after a single icv injection of either FGF-1 or vehicle, we identified >20 cell clusters and >900 cell type-specific differentially expressed genes (p<0.01). Among these potential targets of the MBH response to FGF-1 in diabetic mice, we found that FGF-1 treatment robustly increased the proportion of newly formed oligodendrocytes, a response that can be predicted to yield new mature oligodendrocytes and associated myelination of axons. These preliminary results demonstrate that transcriptional identification and characterisation of icv FGF-1-responsive cell clusters may both advance our understanding of brain control of glucose homeostasis and identify novel targets for diabetes prevention and treatment.

Disclosure

M.A. Bentsen: None. D. Rausch: None. J. Scarlett: None. K.M. Alonge: None. P.N. Timshel: None. Z. Mirzadeh: None. A. Secher: Employee; Self; Novo Nordisk A/S. Stock/Shareholder; Self; Novo Nordisk A/S. Employee; Spouse/Partner; Gubra. R. Jorgensen: Employee; Self; Novo Nordisk Inc.. T. Pers: None. M.W. Schwartz: Consultant; Self; Novo Nordisk A/S. Research Support; Self; Novo Nordisk A/S.
The Cell Junction Protein Afadin Negatively Regulates Insulin Action and Modulates Adipose Tissue Function
Authors: LUNDH MORTEN , ISIDOR MARIE S. , PLUCINSKA KAJA , SHAMSI FARNAZ , PETERSEN PATRICIA , TSENG YU-HUA , EMANUELLI BRICE
Published: June 2018

Adipose tissue dysfunction is one of the main culprits triggering metabolic imbalance associated with obesity. We identified the cell junction protein Afadin as one of the most highly phosphorylated proteins in response to insulin/IGF-1 stimulation using quantitative phospho-proteomic analysis in pre-adipocytes, and investigated the role of Afadin in mediating insulin action and in adipocyte biology. We found that phosphorylation of Afadin occurred in response to insulin in both pre- and mature adipocytes obtained from mice and human subjects. In addition, Afadin phosphorylation was observed in mouse adipose tissues following insulin injection, and after re-feeding, indicating physiological relevance. In obese mice, however, brown fat insulin-induced phosphorylation of Afadin was abolished. Surprisingly, glucose and insulin tolerance were improved in Fat-specific Afadin knock-out (KO) mice on a high fat diet, despite similar body weight gain and adipose mass as wild type controls. Furthermore, ablation of Afadin in vitro did not affect adipogenesis. Yet, prolonged insulin-induced phosphorylation of Akt was observed in KO adipocytes, and was associated with increased insulin-mediated fatty-acid uptake. Finally, phosphorylation of Afadin regulates the docking of several proteins, which may be involved in the regulation of insulin action and adipocyte metabolism. Thus, Afadin is a novel bona fide substrate of the insulin signaling pathway, and is involved in the regulation of insulin action and metabolism.

Disclosure

M. Lundh: None. M.S. Isidor: None. K. Plucinska: None. F. Shamsi: None. P. Petersen: None. Y. Tseng: Other Relationship; Self; Chugai Pharmaceutical Co., Ltd.. Research Support; Self; MedImmune. B. Emanuelli: None.