bluuebunny/crossref_metadata_2025_split huggingface dataset:
262-OR: INAD 012-776 Pilot Study: Allogeneic “Neo-Islets,” Composed of Mesenchymal Stem and Islet Cells, Are Immune Protected and Dose-Dependently and Durably Control Autoimmune T1DM in Pet Dogs
Authors: WESTENFELDER CHRISTOF , CHOWDHURY SABIHA S. , GOOCH ANNA 
Published: June 2019

We demonstrated that allogeneic, ip administered “Neo-Islets” (NIs), aggregates of cultured islet cells (ICs) and immune- and cyto-protective Adipose Derived Stem Cells (ASCs), reestablished durable normoglycemia through omental engraftment and splenic and omental upregulation of Tregs, in autoimmune T1DM NOD mice without immunosuppressive agents (SCTM 2017;6:1631). Comparable euglycemia was achieved with dog- or human-derived NIs in STZ-diabetic NOD/SCID mice. Here we update our report on an FDA supervised study using NI therapy in diabetic pet dogs.
Insulin dependent, diabetic pet dogs, ≤12 kg, were included; 7 enrolled; 5 treated; and 4 followed for ≥6 mos. Pre-treatment serum samples were tested for islet autoantibodies. Comorbidities and blood glucose levels were treated. NIs were given once ip in a dose escalation protocol (2.5x10e5 to 5x10e5/kg bw). Blood glucose levels, insulin need and antibody responses were monitored.
Prior to treatment 3 dogs had islet autoantibodies indicating autoimmunity, and 1 dog had no islet autoantibodies, potentially indicating insulin resistance. NIs appear to engraft, redifferentiate and physiologically produce insulin, and are neither rejected by auto- or allo-immune attacks, as evidenced by (a) an absent IgG response to the administered NIs, and (b) progressively, durably (≥ 12 mos) and dose-dependently improved glycemic control, achieved with a 50% reduction in daily insulin need paralleled by a fall in HbA1c in one dog from 12.1 to 6.8%. One dog was not compliant with keeping blood glucose levels controlled in the first few weeks post treatment and failed to respond.
We conclude that this novel therapy may have significant translational relevance to human T1DM.
We thank site PIs Drs. R. Sellon, WSU, Pullman, WA, N. Loy Son and J. Fisher, Veterinary Specialty Hospital, San Diego, CA, for their excellent collaboration.

Disclosure
C. Westenfelder: Consultant; Self; SymbioCellTech. S.S. Chowdhury: Employee; Self; SymbioCellTech. A. Gooch: Board Member; Self; SymbioCellTech.
265-LB: Mitochondria-Targeted DNA Repair Glycosylase OGG1 Protects Against HFD-Induced Obesity and Insulin Resistance in OGG1 Deficient Mice
Authors: YUZEFOVYCH LARYSA , RACHEK LYUDMILA 
Published: June 2019

Mitochondrial DNA (mtDNA) damage heightens mitochondrial oxidative stress, which is very critical for the development of insulin resistance. OGG1 (8-oxoguanine DNA glycosylase-1) is a DNA glycosylase mediating the first step in the base excision repair which removes 7,8-dihydro-8-oxoguanine (8-oxoG) and repairs oxidized nuclear and mtDNA. Previous studies showed that Ogg1 deficiency results in an increased susceptibility to high fat diet (HFD)-induced obesity, metabolic dysfunction and IR in mice, suggesting a crucial role of Ogg1 in glucose metabolism. Here we performed studies in vivo using Ogg1 knockout and overexpressing genetic models: 1)Ogg1 -/- (KO); 2)KO mice with mitochondrial targeted hOGG1 (hOGG1, subunit 1-α; KO/Tg); 3) wild type (WT) mice. Male mice of all three genotypes were fed either high fat diet (HFD) or control low fat diet (LFD) for 16 weeks, and obesity and insulin resistance phenotypes were evaluated. Our results demonstrated that HFD-fed KO mice displayed increased obesity and whole-body insulin resistance phenotype compared to both WT and KO/Tg mice fed with HFD. Additionally, we have utilized a translational strategy of targeting the recombinant hOGG1 into mitochondria using a TAT-protein transduction system. KO mice were fed HFD and vehicle or MTS-hOGG1-TAT protein was administered intraperitoneally twice weekly for 9 weeks. Consistent with results obtained for genetic models, KO mice injected with MTS-hOGG1-TAT showed significantly lower blood glucose levels and insulin resistance phenotype compared to mice injected with vehicle. Our findings suggest that protecting mtDNA from damage might be crucial to prevent insulin resistance. Furthermore, our data provides strong rationale to use TAT-delivery of DNA repair enzymes into mitochondria as a new translational approach for treatment of insulin resistance.

Disclosure
L. Yuzefovych: None. L. Rachek: None.

Funding
American Heart Association (16GRNT29150012 to L.R.)
267-OR: Cardiomyocyte Insulin Resistance Manifests on High-Glucose Fibroblast-Derived Extracellular Matrix
Authors: NIEUWOUDT STEPHAN , WANG XINMING , SENAPATI SUBHADIP , PARK PAUL , SENYO SAM 
Published: June 2019

The primary cause of death in diabetes is heart failure. Growing evidence suggests that remodeling of the physical microenvironment of the heart drives cardiomyocyte pathology, independent of hypertension and coronary artery disease. Several studies have thus far provided evidence that the extracellular matrix (ECM) changes with diabetic peripheral complications by vascular obstruction and fibrotic hypertension. Rather than a secondary phenomenon, our in vitro data implicates fibroblast derived ECM as a direct regulator of cardiomyocyte insulin sensitivity. Using published methods, we observe that ECM derived from plated primary neonatal cardiac fibroblasts is structurally altered when produced in low- (5mM) versus high-glucose (25mM), with respect to density indicative of collagen crosslinking (Aniline blue staining) and increased stiffening (Atomic Force Microscopy). Most importantly, plating primary cardiomyocytes on high-glucose (vs. low-glucose) fibroblast derived matrix reduces insulin sensitivity (Akt/PKB phosphorylation). We also observe reduced Focal Adhesion Kinase (FAK) activation and c-Jun N-terminal kinase (JNK) inactivation within this in vitro platform; pathways which have been shown to regulate insulin sensitivity via integrin binding to ECM proteins. The cardiac fibroblast derived matrix model serves as a high throughput in vitro platform, which we have thus far used to establish the direct relationship between ECM deposition and insulin sensitivity. Future work will use the platform to dig into the underlying mechanisms implicating ECM remodeling and sensing in the pathogenesis of diabetic cardiomyopathy and heart failure.

Disclosure
S. Nieuwoudt: None. X. Wang: None. S. Senapati: None. P. Park: None. S. Senyo: None.

Funding
Case Western Reserve University (RES221997); Wallace H. Coulter Foundation; National Heart, Lung, and Blood Institute (T32HL134622-02)
269-OR: miR-409-3p Regulates Angiogenesis Brown Fat Adiposity and Insulin Resistance
Authors: BECKER-GREENE DAKOTA R. , LI HAO , WU WINONA , OZDEMIR DENIZHAN , HOLLAN IVANA , FEINBERG MARK W. , ICLI BASAK 
Published: June 2019

White and brown adipose tissues are highly vascularized organs, capable of plasticity based on metabolic demands and energy expenditure, which if maladaptive can lead to insulin resistance. Critical gaps remain in our understanding of how angiogenesis impacts adipose tissue dysfunction and overall metabolism. MicroRNAs (miRs) are implicated in the regulation of the angiogenic response to pathophysiological stimuli. Function of miRNAs regulating the angiogenic response in diet-induced insulin resistance remains poorly understood. Using a miRNA-Seq approach, we identified that miR-409-3p expression was significantly increased in endothelial cells (ECs) of brown adipose tissue (BAT) of diet-induced obese (DIO) mice and in human diabetic plasma samples compared to nondiabetic patients. Overexpression of miR-409-3p markedly inhibited EC growth and migration, whereas miR-409-3p inhibition had the opposite effects. miR-409-3p targets the 3’UTR of Zinc Finger E-box binding Homeobox 1 (ZEB1). Overexpression of miR-409-3p decreased ZEB1 expression in ECs and in human subcutaneous white adipose tissue (sWAT). SiRNA knock down of ZEB1 expression in ECs phenocopied the effects of miR-409-3p overexpression and significantly decreased cell proliferation and migration. In addition, 3T3-L1 cells co-cultured with supe harvested from ECs deficient in miR-409-3p had increased expression of brown fat markers (UCP1, Cidea, PGC1-α) by RT-qPCR and Western blot analyses. Systemic intravenous delivery of LNA-anti-miR-409-3p inhibitor to DIO mice significantly increased angiogenesis by CD31 staining, accompanied by higher UCP-1 in BAT and sWAT by RT-qPCR, Western blot and immunohistochemistry analyses, while improving glucose tolerance and overall metabolism. miR-409-3p plays a key regulatory role between impaired angiogenesis in diet-induced obesity and adipose tissue dysfunction, an effect that could be exploited for therapeutic intervention in obesity-induced insulin resistance.

Disclosure
D.R. Becker-Greene: None. H. Li: None. W. Wu: None. D. Ozdemir: None. I. Hollan: None. M.W. Feinberg: None. B. Icli: None.

Funding
American Diabetes Association (1-16-JDF-046 to B.I.)
273-LB: Leukocyte-Endothelial Interaction Is Reduced by Insulin-Induced Downregulation of Endothelial CXCR4
Authors: KUNKEMOELLER BRITTA , RATHJEN THOMAS , PATTI JAMES C. , LI QIAN , PARK KYOUNGMIN , KING GEORGE L. , RASK-MADSEN CHRISTIAN 
Published: June 2019

An activated, pro-inflammatory endothelium is a key feature of obesity and type 2 diabetes and may contribute to adipose tissue inflammation, atherosclerosis and tumor formation. We have previously published a striking 4-fold increase in leukocyte adhesion to endothelium in mice with endothelial cell knockout of the insulin receptor. Here, we searched for insulin-regulated genes involved in leukocyte-endothelial cell interaction by performing RNA sequencing in primary human endothelial cells. Among genes with the largest change in expression after insulin treatment was C-X-C motif chemokine receptor 4 (CXCR4), which was downregulated by 54%. This reduction was completely prevented by pretreatment with wortmannin but not by U0126, an inhibitor of MAPK signaling. In CD31+ cells isolated by fluorescence-activated cell sorting of enzymatically dissociated tissue, CXCR4 mRNA in was 77% higher in mice with diet-induced obesity compared to lean controls and 37% higher in db/db mice compared to db/+ controls, consistent with upregulation of CXCR4 in endothelial cell insulin resistance. Stromal-derived factor-1 (SDF-1), the ligand for CXCR4, increased leukocyte adhesion to cultured endothelial cells. Insulin treatment of endothelial cells prevented 80% of this increase which in turn was abrogated by plerixafor, a clinically approved CXCR4 antagonist, or deletion of CXCR4 by gene editing. To study gain of endothelial cell insulin signaling in vivo, we examined transgenic mice with endothelial overexpression of insulin receptor substrate-1 (IRS-1). In vivo microscopy of mesenteric venules showed an increase in leukocyte rolling and adhesion after intravenous injection of SDF-1 in wildtype mice which was reduced by 83% in transgenic mice. We conclude that endothelial cell insulin signaling limits leukocyte-endothelial cell interaction through downregulation of CXCR4 and that targeting endothelial CXCR4 may protect against inflammation in insulin resistant states.

Disclosure
B. Kunkemoeller: None. T. Rathjen: Employee; Self; Bayer AG. J.C. Patti: None. Q. Li: None. K. Park: None. G.L. King: Research Support; Self; Sanofi. C. Rask-Madsen: None.
274-OR: Beta-Cell Dysfunction Is Associated with Impaired Insulin-Mediated Suppression of Glucagon in Obese Hispanic Americans
Authors: CHEN XI , MALDONADO CORCHADO ENRIQUE R. , MEROVCI AURORA , CASE EMINA , TRIPATHY DEVJIT , DEFRONZO RALPH A. 
Published: June 2019

Recent studies have demonstrated an increase in the glucagon:insulin ratio in lean versus obese subjects. We assessed insulin-mediated suppression of plasma glucagon levels in 260 obese Hispanic Americans (59 NGT, HbA1c= 5.1±0.1%, BMI= 32.6±0.7; 109 Prediabetes, HbA1c= 5.8±0.1%, BMI= 33.8±0.5; 92 T2D, HbA1c= 7.3±0.2%, BMI= 34±0.5) with varying glucose tolerance. Subjects participated in a 2-h OGTT with measurement of plasma glucose, insulin, glucagon concentrations. During the OGTT, we quantitated insulin-mediated suppression of plasma glucagon concentration, Matsuda Index of insulin sensitivity, and the insulin secretion/insulin sensitivity (disposition) index using plasma C-peptide concentration. There was no difference in fasting plasma glucagon level between obese subjects with normal glucose tolerance (NGT), prediabetes, and type 2 diabetes (T2D). However, there was a progressive decrease in fasting glucagon:insulin ratio from NGT to prediabetes to T2D (8.71±0.72 vs. 6.92±0.53 vs. 5.66±0.41, P&lt;0.01). During OGTT at 60 min, glucagon:insulin ratio increased from NGT to T2D (0.93±0.09 vs. 0.89±0.07 vs. 2.33±0.20, P&lt;0.001), and the suppression of plasma glucagon (compared to basal value) was markedly decreased (10.3±3.4% vs. 9.3±1.9% vs. 2.5±2.8%, P&lt;0.05). At 120 min (OGTT), glucagon:insulin ratio was increased in T2D versus NGT (1.28±0.17 vs. 2.0±0.20, P&lt;0.01). The glucagon:insulin ratio was closely correlated with whole body insulin sensitivity and β-cell function during the fasting state (Matsuda Index, r= 0.51, P&lt;0.001; Disposition Index, r= 0.30, P&lt;0.001) and with Disposition Index during the OGTT at 60 and 120 min (r= -0.41, P&lt;0.001; r= -0.21, P&lt;0.01), respectively. Collectively, these findings indicate that insulin-mediated suppression of glucagon secretion is impaired with increasing severity of glucose intolerance in obese Hispanic Americans and is closely associated with insulin resistance and β-cell dysfunction.

Disclosure
X. Chen: None. E.R. Maldonado Corchado: None. A. Merovci: None. E. Case: None. D. Tripathy: None. R.A. DeFronzo: Advisory Panel; Self; AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Elcelyx Therapeutics, Inc., Intarcia Therapeutics, Inc., Janssen Pharmaceuticals, Inc., Novo Nordisk Inc. Research Support; Self; AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Janssen Pharmaceuticals, Inc., Merck & Co., Inc. Speaker's Bureau; Self; AstraZeneca, Novo Nordisk Inc.
275-LB: Connectome Predictive Modeling May Identify Brain Connectivity Signatures to Help Predict Who Will Benefit from Low-Calorie Diet
Authors: GROSKREUTZ DEREK , LAM WAI , LACADIE CHERYL , ELSHAFIE AHMED , HWANG JANICE J. , SEO DONGJU , SAVOYE MARY , SINHA RAJITA , CONSTABLE TODD , SHERWIN ROBERT , BELFORT-DEAGUIAR RENATA 
Published: June 2019

While studies have investigated changes in brain connectivity following diet and weight loss, it has not yet been explored whether baseline brain connectivity can be used to predict weight loss during diet or to identify individual subjects for which a particular diet may be effective. The following study uses Connectome-based predictive modeling (CPM), a data-driven analysis approach for producing predictive models of individual brain-behavior relationships from brain connectivity data using cross-validation, to help identify individual participants who will lose weight after an 8-week low calorie diet. 16 Healthy OB subjects (10F/6M, age 44.4±8 years, BMI 32.7±2) and 9 T2DM subjects (5F/4M, age 48±9, BMI 33.9±2) underwent functional MRI (fMRI) during a hyperglycemic-euglycemic clamp. Blood-oxygen-level-dependent (BOLD) brain activity was assessed while subjects viewed food (high-calorie, low-calorie) pictures and non-food images. The study was repeated after 8 weeks of a reduced calorie diet. CPM was applied to BOLD activity during hyperglycemia and euglycemia. Analyses show that models built on individual subject brain connectivity matrices averaged between hyperglycemia and euglycemia before an 8-week low calorie diet are able to predict 50.2% and 53.4% of the variance in BMI and weight loss (kg), respectively (p&lt;0.05). While models built on brain connectivity matrices during euglycemia (similar to the post-prandial state), but not hyperglycemia predicted 42.2% of the variance in BMI change (p=0.017). These results suggest the possibility that individual brain connectivity signatures may be used to identify subjects for whom a low calorie diet might be an effective weight loss strategy, while also revealing the relevant functional connections associated with weight loss success.

Disclosure
D. Groskreutz: None. W. Lam: None. C. Lacadie: None. A. Elshafie: None. J.J. Hwang: None. D. Seo: None. M. Savoye: None. R. Sinha: None. T. Constable: None. R. Sherwin: Other Relationship; Self; ICON plc., IQVIA, MannKind Corporation. R. Belfort-DeAguiar: Research Support; Self; Silver Palate Kitchens, Inc.

Funding
National Institutes of Health
277-OR: Lean Tissues as Novel Determinants of Pear vs. Apple Body Shape and Metabolic Health in Humans
Authors: KARASTERGIOU KALYPSO 
Published: June 2019

Epidemiological studies have established that pear-, compared to apple-, body shape is associated with lower cardiometabolic risk. Up to date, body shape has been considered equivalent to fat distribution and significant research efforts investigated the biology of different fat depots and their metabolic effects. We undertook a systematic analysis of body and tissue compartments in participants in the NHANES 1999-2006 cycles to search for components of body shape, other than fat. As expected, among adults 20-69 years old, those with pear-body shape (defined as waist-to-thigh ratio &lt; 1.65 - bottom quartile) had higher leg fat mass compared to those with apple-body shape (waist-to-thigh ratio &gt; 1.95 - top quartile) by 1,967 g (95% CI 1,801-2,135 g), after adjustment for sex, ethnicity, age and BMI. Surprisingly it was also noted that subjects with pear-body shape had slightly higher leg bone mineral content (by 42 g, 95% CI 30-52 g) and significantly higher leg lean mass (by 1,218 g, 95% CI 1,072-1,365 g). Similar results were observed when analysis was restricted to subjects &lt; 40 years old, women only, or in specific ethnicities. In multiple regression models, leg lean mass (or leg/total lean mass ratio, i.e., lean mass distribution) was an independent predictor of lower systolic blood pressure, HbA1c and CRP after adjustment for sex, age, ethnicity, BMI, and leg fat mass (or leg/total fat mass, i.e., fat distribution), but not a predictor of indices of insulin sensitivity (fasting insulin, triglycerides, HDL-cholesterol).
In conclusion, fat distribution is not the sole determinant of pear-body shape in humans. Rather, coordinated differences in fat and lean mass drive metabolic health and suggest a systemic developmentally-driven pattern of lower body expansion.

Disclosure
K. Karastergiou: None.

Funding
National Center for Advancing Translational Sciences (KL2TR001435)
28-LB: Diabetic Retinopathy Is an Additional Risk Factor for Hemodialysis Induction and Mortality in Type 2 Diabetic Patients
Authors: MORIYA TATSUMI , MATSUBARA MADOKA , OUCHI MOTOSHI 
Published: June 2019

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

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

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

Disclosure
S. Jain: None. D.K. Tosh: None. M. Reitman: None. K.A. Jacobson: None.