bluuebunny/crossref_metadata_2025 huggingface dataset:
Generation of Insulin Expressing Cells in Mouse Small Intestine by Pdx1, MafA, and BETA2/NeuroD
Authors: HYUN LEE SO , KIM JIWON , YOON KUN-HO
Published: June 2018

To develop the beta-like cells for diabetes therapy, adult stem cells have been identified in various tissues and studied for their conversion into beta cells. Pancreatic progenitor cells are derived from the endodermal epithelium and formed in a similar manner as gut progenitor cells. Here, we generated insulin-expressing cells from the intestinal epithelial cells that induced many of the specific pancreatic transcription factors using adenoviral vectors carrying three genes: Pdx1, MafA, and BETA2/NeuroD (PMB). By direct injection into the whole intestine, adenoviruses (Ad) were successfully delivered to the intestine. After virus injection, we could confirm that the small intestine of the mouse was appropriately overexpressed with the Ad-Pdx1 and triple Ad-PMB. Four weeks after the injection, insulin mRNA was expressed in the small intestine, and the insulin gene expression was induced in Ad-Pdx1 as well as Ad-PMB compared to control Ad-GFP. In addition, the conversion of intestinal cells into insulin-expressing cells was detected in parts of the crypts and villi located in the small intestine.

In conclusion, these data indicated that Pdx1, MafA, and BETA2/NeuroD facilitate the differentiation of mouse intestinal cells into insulin-expressing cells. The small intestine is an accessible and abundant source of surrogate insulin-producing cells.

Disclosure

L. So Hyun: None. J. Kim: None. K. Yoon: None.
Evaluation of CFTR Expression and Localisation in Human Pancreas
Authors: MAHESHWARI RASHMI R. , JONES CLAIRE J. , SHAW JAMES A.M. , WHITE MICHAEL G.
Published: June 2018

Introduction: Cystic fibrosis-related diabetes (CFRD) is the most common co-morbidity in people with CF, occurring in 40-50% of adults. Whilst it is established that β-cell dysfunction in cystic fibrosis (CF) leads to diabetes, the mechanism by which the CF transmembrane conductance regulator (CFTR) channel influences insulin secretion remains debated. Currently, three major hypotheses have been proposed: 1. Intrinsic CFTR-dependent pathways of insulin secretion 2. Pancreas-extrinsic CFTR defects 3. Remodeling of islets following loss of exocrine tissue due to inflammation. Since the contribution of each to the pathogenesis of CFRD remains largely unknown, we sought to determine CFTR localisation within human pancreas using novel and highly sensitive approaches.

Methods: Expression of chromogranin A (ChrA), CFTR and keratin 19 (K19) was assessed by immunofluorescence (IF) staining of tissue obtained from deceased donors without diabetes (n=10, age: 23-71 years). Two CFTR antibodies (ab576 and ab590) obtained from the CF Foundation were used to confirm specificity, with ductal and endocrine cells determined by K19 and ChrA respectively. CFTR RNA expression was determined by RNAscope® in situ hybridisation (ISH) and combined with either ChrA or insulin immunohistochemistry (IHC).

Results: IF staining of pancreatic tissues indicated co-localisation of CFTR in K19+ ductal cells, but not in ChrA+ endocrine cells. These observations were confirmed by combined RNAscope® ISH and IHC (ChrA and insulin), which demonstrated the absence of CFTR RNA in human islets.

Conclusion: Employment of these highly sensitive techniques has demonstrated absence of CFTR within normal β-cells or any other islet endocrine cell types. This is in line with recent observations in isolated human islets. We conclude that CFTR abnormalities do not directly impact beta-cell function and that CFRD is mediated by factors extrinsic to the pancreatic endocrine compartment.

Disclosure

R.R. Maheshwari: None. C.J. Jones: None. J.A.M. Shaw: Advisory Panel; Self; Novo Nordisk A/S. M.G. White: None.
Quantitative Proteomic Analysis on Human Islets—New Markers of Cellular and Metabolic Dysfunction
Authors: ASSUNTA CEFALO CHIARA MARIA , MEZZA TERESA , KULKARNI ROHIT , GIACCARI ANDREA
Published: June 2018

The pathogenesis of type 2 diabetes is characterized by a progressive β-cell dysfunction resulting in both quantitative and qualitative loss of insulin secretion. However, the molecular mechanisms underlying this progressive functional loss are still unknown. The aim of the study is to highlight changes in the proteome of pancreatic human islets that can anticipate and eventually predict diabetes’ onset. High performance liquid chromatography mass spectrometry (HPLC-MS) analysis was applied to islets isolated by laser capture microdissection (LCM) from human samples of both diabetic and no diabetic subjects, underwent to duodeno-cefalopancreatectomy for extra-pancreatic, low grading tumors. The subjects were classified on the basis of glucose tolerance assessed by an OGTT before surgery in normal tolerant (NGT, n:7), glucose intolerant (IGT, n:5) and diabetic subjects (T2DM, n:2). Qualitative and quantitative analysis were performed to detect differential protein expression among the 3 study’s groups. Sixty-seven proteins were found to be differentially regulated in diabetic subjects compare to NGT, with 29 upregulated and 38 downregulated proteins, while 95 proteins were differentially expressed in IGT compare to NGT with 49 upregulated and 46 downregulated. These proteins are mainly involved in cellular and metabolic processes. In particular IGT and DM, compared to NGT, showed a lower expression of proteins involved in cell proliferation such as PURA and NAP1L1, as well as proteins involved in the insulin cleavage process, like ERO1B. While other proteins involved in endoplasmic reticulum stress, CASP14, ERP27 and PDIA3, resulted upregulated in both IGT and DM compare to NGT. Our data suggest that metabolic and cellular processes of islet cells are already dysregulated in patients with impaired glucose tolerance, highlighting some proteins as potential early markers of β-cell dysfunction and novel therapeutic targets to slow the progression to type 2 diabetes.

Disclosure

C. Cefalo: None. T. Mezza: None. R. Kulkarni: None. A. Giaccari: Advisory Panel; Self; Sanofi-Aventis. Speaker's Bureau; Self; AstraZeneca, Merck Sharp & Dohme Corp., Amgen Inc..
Type 1 Diabetes Development Is Modulated by Select Lipid Signaling Derived from Beta Cells and Immune Cells
Authors: NELSON ALEXANDER , PARK MARGARET A. , CHALFANT CHARLES , RAMANADHAM SASANKA
Published: June 2018

Type 1 diabetes (T1D) is a consequence of autoimmune destruction of β-cells and we find that the Ca2+-independent phospholipase A2β (iPLA2β) is induced in β-cells and immune cells and mitigation of iPLA2β attenuates β-cell death and T1D incidence. Activation of iPLA2β leads to generation of pro-inflammatory or anti-inflammatory bioactive lipids and we previously reported that iPLA2β activation favors M1 pro-inflammatory macrophage phenotype. Here, we utilized UPLC ESI-MS/MS protocols to assess the lipid profile generated by macrophages as a means to identify select lipids that are associated with T1D development. Macrophages were treated with DMSO (vehicle), IFNγ+LPS (to induce M1 polarization), or IL-4 (to induce M2 polarization) and at 16h, the media was collected and processed for assessment of eicosanoid class of bioactive lipids. Comparison of spontaneous diabetes-resistant C57BL/6J and spontaneous diabetes-prone NOD mice revealed that nearly all pro-inflammatory eicosanoids were elevated in NOD during the prediabetic phase, relative to C57. Theabundances increased in the NOD between 4-8 and decreased by 15 weeks, correlating with onset of insulitis. These findings are consistent with a higher inflammatory status in the NOD vs. C57. To assess contribution of iPLA2β to the lipid profile, macrophage responses in NOD were compared with those in NOD.iPLA2β-/+. Such analyses revealed that production of select pro-inflammatory and anti-inflammatory lipids were decreased and increased, respectively, from macrophages and islets of NOD.iPLA2β-/+, relative to age-matched NOD-WT, and this correlated with reduced T1D incidence in the NOD.iPLA2β-/+. We further find that iPLA2β-derived lipids (iDLs) play a critical role in alternative splicing events that favor generation of pro-apoptotic variants of several factors (Bcl-x, capsase-9, and RAGE). These findings raise the possibility that iDLs are candidates for targeting to counter T1D development.

Disclosure

A. Nelson: None. M.A. Park: None. C. Chalfant: None. S. Ramanadham: None.
Increased Obesity Is Causal for Increased Inflammation—A Mendelian Randomisation Study
Authors: ZUYDAM NATALIE VAN , WIELSCHER MATTHIAS , MCCARTHY MARK , JARVELIN MARJO-RIITTA
Published: June 2018

Inflammation is endemic to obesity and type 2 diabetes (T2D) but it is unclear if it is a cause or a consequence. If the latter, then inflammation may increase the risk of obesity and diabetes complications. Large genome-wide association studies (GWAS) of inflammation and cardiometabolic traits can be used to investigate causal mechanisms. Summary statistics were assembled from GWAS of 49 cytokine (N=840-80,000) and 20 cardiometabolic traits (N=2,447-465,333). Shared genetic background was measured using LD score regression. Polygenic scores (PGS) were generated from GWAS data for 49 cytokines (representing inflammation) and 20 cardiometabolic traits and used in two-way Mendelian Randomisation (MR) analyses. Causal relationships were identified using summary statistics for all traits, and individual data (UK Biobank, N=465,333) to test the association of cytokine PGS with body mass index (BMI) and T2D. LD score regression analysis demonstrated that BMI and T2D shared a genetic background with acute phase cytokines. However, MR analyses showed that genetically-driven variation in cytokine levels were not associated with increased BMI or T2D risk. This argues against a causal role for inflammation (as represented by these cytokines) in the development of obesity and T2D. In contrast, genetically driven obesity was associated with increased inflammation: a PGS for increased BMI was associated with increased C-reactive protein (Beta [95% CI], 0.40 [0.34,0.46], p=1.3×10-36) and activated plasminogen inhibitor (Beta [95% CI], 0.39 [0.19, 0.59], p=7.8×10-5) levels. Genetically-driven variation in cytokine levels were causally linked with chronic kidney disease in subjects with T2D: fibroblast growth factor 2 (OR[95% CI], 0.58[0.46,0.72], p=1.8×10-6) and interleukin 13 (OR[95% CI], 1.28[1.15,1.43], p=7.0×10-6) We conclude that chronic inflammation is a consequence, rather than a cause, of T2D and obesity, and may contribute to some of their complications.

Disclosure

N. van Zuydam: None. M. Wielscher: None. M. McCarthy: Research Support; Self; Eli Lilly and Company, Takeda Pharmaceuticals U.S.A., Inc., Novo Nordisk A/S. Advisory Panel; Self; Novo Nordisk A/S, Pfizer Inc.. Research Support; Self; Pfizer Inc., Sanofi, Boehringer Ingelheim GmbH, AstraZeneca, Merck Sharp & Dohme Corp.. M. Jarvelin: None.
AER Is a More Sensitive Predictive Factor than eGFR for CVD in Patients with Type 2 Diabetes—Results from a Perspective Study in Beijing Communities
Authors: ZHANG XUE LIAN , YUAN SHEN YUAN ,   BCDS GROUP
Published: June 2018

Objective: The predictive values of AER and eGFR for CVD outcomes in T2DM patients have not been well documented. In this study, the effects of AER and eGFR on outcomes of CVD were analyzed in a population of T2DM patients.

Methods: 1914 T2DM patients were enrolled in this 8 -year prospective study in Beijing Communities. The patients’ serum creatinine levels were lower than 176.8μmol/l. The risk of CVD onset was assessed according to CKD staging, which was categorized by urine albumin excretion (AER; mg/d) and estimated GFR (eGFR; ml min −11.73 m−2). The effects of AER and eGFR on risk of cardiovascular disease onset in type 2 diabetes after eight years’ prevention were also analyzed.

Results: The mean age and BMI of patients were 66.4 ± 10.1 years and 25.5 ± 3.5 kg/m2, respectively; the duration of diabetes was 5.7 to 17.8 years. The proportion of patients with hypertension and smokers were 71.11% and 16.84%. During the follow-up period (median 6.8 years), 71 CVD events occurred. At baseline, those with AER≥300 mg/d and co-existing eGFR 60-89 ml min −11.73 m−2 or <60 ml min −1/1.73 m−2 showed increased risk for CVD onset when compared with ’no CKD’. No increased risk was observed for patients with AER <30 mg/d and eGFR <60 ml min −11.73 m−2. The increased CVD risk was observed in patients progressed to AER ≥30mg/d during follow-up period, whereas patients who progressed to eGFR <90 ml min −11.73 m−2 alone showed no increased CVD risk. During the follow-up period, 8.7% patients with microalbuminuria (n=166) and 1.8% patients with overt nephropathy (n=34) reversed to normoalbuminuria or microalbuminuria.

Conclusions: AER is a more sensitive predictor than eGFR for CVD onset in T2DM patients. DKD, even patients with overt nephropathy can be reversed after multi-factorial intervention in a part of patients.

Disclosure

X. Zhang: None. S. Yuan: None.
Association of Maternal and Offspring Genetic Risk for Type 2 Diabetes with Offspring Birth Weight among African-Ancestry Populations
Authors: RAHMAN MOHAMMAD L. , SHRESTHA DEEPIKA , WORKALEMAHU TSEGASELASSIE , WU JING , ZHANG CUILIN , TEKOLA-AYELE FASIL
Published: June 2018

Background: Mother’s genetic risk for type 2 diabetes (T2D) may influence intra-uterine glycemia leading to increased birth weight. However, the U-shaped relation between offspring birthweight and later-life T2D risk suggests that relations of offspring’s T2D genetic susceptibility with birthweight may be U-shaped.

Methods: We investigated the effects of maternal and offspring genetic risk for T2D on birthweight using genotype data from 949 mother-offspring pairs of African ancestry from the Hyperglycemia Adverse Pregnancy Outcome study. We generated a genetic risk score for T2D (dGRS) for mothers and offspring as the weighted sum of 90 T2D risk alleles. Offspring dGRS was separately calculated for birth weight-reducing and -increasing alleles based on the direction of correlation between a T2D-risk allele and birthweight in our data. We used linear regression adjusting for maternal and offspring covariates and the proportion of African ancestry to estimate the change in birthweight by quartiles of maternal and offspring dGRS.

Results: Changes in birthweight across increasing quartiles of mother’s dGRS were 0 (reference), 83.3g (95% CI: 6.7, 160.0), 102g (95% CI: 25.1, 179.5), and 92.9g (95% CI: 12.7, 173.1) (P-trend=0.042). In contrast, changes in birthweight across increasing quartiles of offspring dGRS based on birthweight-reducing alleles were 0, −105.7g (95% CI: −180.9, −30.5), −105.5g (95% CI: −180.4, −30.5), and −143.2g (95% CI: −218.6, −67.8) (P-trend=0.001), whereas those of dGRS based on birthweight-increasing alleles were 0, 15.3g (95% CI: −60.7, 91.3), 113.3g (95% CI: 38.1, 188.6), and 142.4g (95% CI: 66.8, 218.1) (P-trend=0.0002).

Conclusion: Maternal and offspring genetic risk for T2D exhibited a differential association with birth weight. Further characterization of individual alleles in a larger dataset will enhance our understanding of the causal roles of fetal and maternal genetic determinants of T2D on fetal growth.

Disclosure

M.L. Rahman: None. D. Shrestha: None. T. Workalemahu: None. J. Wu: None. C. Zhang: None. F. Tekola-Ayele: None.
The Burden of Uncontrolled Glycemia—A Systematic Literature Review
Authors: FERNANDES GAIL , IFF JOEL , OVCINNIKOVA OLGA , NASSIM MARIA , SUN LUCY , RICHARDS MARGARET S. , RAJPATHAK SWAPNIL
Published: June 2018

The objective of this systematic literature review (SLR) was to understand the consequences of uncontrolled glycemia in patients with type 2 diabetes mellitus (T2DM) in terms of mortality, macro- or microvascular complications, as well as economic burden in terms of healthcare resource utilization (HCRU) and cost. A predefined search strategy was used in Medline, EMBASE, Cochrane CENTRAL, EconLit and NHS-EED, limited to the last five years, according to PRISMA guidelines. In terms of mortality and clinical data, twenty-two studies met our inclusion criteria. Five studies reported significantly increased risk of mortality for A1C greater than 7.5% (Hazard ratio (HR) ranged from 1.41-13.16). Three studies reported an increased risk of mortality for every 1% increase in A1C above 7% (HR from 1.10-1.40). In one landmark trial (ACCORD), however, comparing intensive (A1C target <6.0%) to standard glycemic control (A1C target 7.0-7.9%), a higher risk of all-cause mortality in the intensive group was reported. Six studies showed that A1C higher than 7% was associated with an increased risk of macrovascular complications (HR / odds ratio (OR) from 1.13-2.26) and three studies reported that every 1% increase in A1C above 7% resulted in an increased risk of different macrovascular complications (HR / OR from 1.05-1.38). Three studies reported higher risk (HR / OR from 1.13-1.40) of microvascular events at A1C higher than 7%. For the economic data, 1 U.S. study reported increased healthcare cost (OR 1.73 (95% CI 1.00-3.01) for T2DM patients with A1C greater than 10%. In another U.S. study, 1% increase in A1C between 7-10.5% was associated with additional diabetes-related pharmacy/medical cost burden of $1586 ($133-$3537) per patient per year. Three non-U.S. studies also reported similar results on the association of A1C and economic burden. Our SLR showed an association between high A1C and increased risk of clinical outcomes, including mortality. Furthermore, poor glycemic control was also associated with increased HCRU and costs.

Disclosure

G. Fernandes: Employee; Self; Merck & Co., Inc.. Employee; Spouse/Partner; Janssen Research & Development. Stock/Shareholder; Self; Merck & Co., Inc.. Stock/Shareholder; Spouse/Partner; Janssen Research & Development. J. Iff: None. O. Ovcinnikova: None. M. Nassim: None. L. Sun: None. M.S. Richards: None. S. Rajpathak: Employee; Self; Merck & Co., Inc..
Accumulation of Endogenous Asymmetric Dimethylarginine Contributes to Diabetic Cardiomyopathy via Endoplasmic Reticulum Stress in Diabetic Rats
Authors: XIONG YAN , HE YULIAN , FANG WEI-JIN , LEI YANPING , LIN YUAN , HE ZHI-MIN
Published: June 2018

Diabetic cardiomyopathy is a major cause of morbidity and mortality among diabetic patients. Endoplasmic reticulum (ER) stress has been involved in diabetic cardiomyopathy. Accumulation of endogenous nitric oxide synthase (NOS) inhibitor asymmetric dimethylarginine (ADMA) is associated with diabetic cardiovascular complications. This study was to determine whether ADMA accumulation contributes to diabetic cardiomyopathy and whether its mechanism is related to ER stress in streptozotocin-induced diabetic rats and cultured rat cardiomyocytes. Cardiac function was measured by M-echocardiography. ER stress was reflected by transcription of glucose-regulated protein78, phosphorylation of eukaryotic initiation factor 2α kinase, splicing of X box-binding protein-1 mRNA and expression of C/EBP homologues protein. Changes of ADMA signal pathway were evaluated by ADMA and nitric oxide contents, NOS and dimethylarginine dimethylaminohydrolase activities, as well as their protein expression. The levels of ADMA in serum and myocardium were significantly elevated in diabetic rats compared to control rats. This elevation was associated with left ventricular diastolic and systolic dysfunction of diabetic rats in comparison with control rats. Furthermore, ADMA accumulation was not only coupled with the disorder of ADMA signal pathway, but also tied to ER stress and oxidative stress in myocardium of diabetic rats. Treatment of diabetic rats with antioxidant N-acetylcysteine could reduce oxidative stress and ER stress, but also ameliorate cardiac dysfunction of diabetic rats. Similar results to diabetic rats were observed in the exogenous NOS inhibitor-treated normal rats and ADMA-incubated cardiomyocytes. These results indicate that endogenous ADMA accumulation plays a pivotal role in the development of diabetic cardiomyopathy, and the underlying mechanism is close related to ER stress and oxidative stress.

Disclosure

Y. Xiong: None. Y. He: None. W. Fang: None. Y. Lei: None. Y. Lin: None. Z. He: None.
Diabetic Ketoacidosis Triggered by Pembrolizumab in a Patient with Bladder Cancer
Authors: SHAMY TANIA AL , AGUASVIVAS MARNIE , SERHAN MUSTAPHA , FOJAS MA.CONCHITINA M.
Published: June 2018

This is a 71-year-old male with history of Hypertension, well controlled type 2 DM on metformin (HbA1c 6.6%), and high grade urothelial bladder cancer treated with transurethral resection of the bladder. Due to cancer progression, he required neoadjuvant chemotherapy followed by Pembrolizumab in July 2017. He presented with fatigue, polyuria and polydipsia in August 2017. He was diagnosed with DKA and managed in the ICU. Workup revealed HbA1c 9%, high GAD-65 antibody (>250 IU/ML), and low C-peptide (0.26 ng/ml). He was stabilized and discharged on insulin. T cell activity is regulated by programmed death ligand-1(PD-L1)/programmed death-1 (PD-1). Tumor cells select specific immune-checkpoint pathways as a mechanism of immune resistance against T cells and evade the antitumor immune response by overexpressing PD-L1. Pembrolizumab is a humanized monoclonal antibody against PD-1 acting as an immune checkpoint inhibitor (ICI). Pembrolizumab is approved for treating metastatic urothelial carcinoma, cisplatin-ineligible cancer or disease progression after cisplatin therapy. ICIs are associated with multi-system immune related adverse events (irAEs) due to the disinhibition of the host immune homeostasis. Most of the irAEs are reversible with supportive treatment, however life-threatening irAEs can occur including severe skin reaction, type 1 DM, hypophysitis, and rhabdomyolysis. In our patient, we suspected pembrolizumab was related to new onset type 1 DM. type 1 DM and DKA were reported in 6 (0.2%) of 2,799 patients on Pembrolizumab in 3 randomized, open-label, and active-controlled clinical trials. There are 12 reported cases of PD-1/PD-L1-associated type 1 DM. The onset of disease from use varied from 1 week to 12 months, with most patients presenting with type 1 DM with DKA. ICIs are breakthroughs in cancer treatment. Clinicians should be aware of their possible irAEs. The development of autoimmune diabetes is rare, but its complications are life threatening if not attended to immediately.

Disclosure

T. Al Shamy: None. M. Aguasvivas: None. M. Serhan: None. M.M. Fojas: None.
IGFBP-2 Ameliorates Vascular Insulin Resistance and Contributes to Cardiometabolism
Authors: LI ZHUO , LV YOU , GUO WEIYING , WANG GUIXIA
Published: June 2018

Insulin-like growth factor binding protein-2 (IGFBP-2) has been implicated in the etiology of insulin resistance. And previous studies have pointed that IGFBP-2 overexpression could prevent age-related hypertension. The present study was aimed to document the modulation of dual vascular actions of IGFBP-2 in human umbilical vein endothelial cells (HUVECs). HUVECs at passage 4-6 were cultured with either 20 μM hydrogen peroxide (H2O2) or 2mM glucosamine (GLU) to induce insulin resistance, which declined the expression of IGFBP-2 in HUVECs and resulted in endothelial nitric oxide (NO) and endothelin-1 (ET-1) production were reduced significantly compare to those control cells (p<0.01), the suppression of NO production was more notable. The insulin-induced phosphorylation of Akt at The3and endothelial NO synthase (eNOS) at Ser1177 were also slightly decreased in IGFBP-2 down-regulated HUVECS. Coherently, insulin-induced phosphorylated eNOS and extracellular signal-related kinase 1/2 (ERK1/2) were 3.5 (p<0.01) and 1.5 (p<0.05) folds increased by an overnight IGFBP-2 treatment, respectively. As the production of ET-1 was mediated by ERK1/2 and the secretion of NO was regulated by eNOS, we suspected that IGFBP-2 played an important role on endothelial diastolic and systolic function, may robustly improved vasodilation more than vasoconstriction in insulin resistant HUVECs.

Disclosure

Z. Li: None. Y. Lv: None. W. Guo: None. G. Wang: None.
Development of DQ-MHC Class II Monoclonal Antibodies—A New, Targeted Therapeutic Approach in Type 1 Diabetes
Authors: LOMBARDI ANGELA , ZHU MING , MORAN THOMAS , KRAUS THOMAS A. , HSIEH MICHAEL , STEVENS CHAD , MURPHY DENISE , DONOVAN CAROL B. , ATKURI KONDALA , MESSING DEAN M. , CHAMOUN JEAN , BROWN THOMAS P. , GUNTAS GURKAN , BENNETT DONALD V. , CONCEPCION ERLINDA , JOHNSON KRISTEN , TOMER YARON
Published: June 2018

Purpose: Type 1 diabetes (T1D) is a disease affecting almost three million Americans; every year 30,000 new cases are diagnosed, 50% of whom are children. There is currently no curative therapy for T1D and the only available treatment is insulin replacement. Substantial recent data demonstrate a strong association between the MHC Class II molecule DQ8 and the development of T1D. Indeed, DQ8 has been shown to present antigenic peptides (such as InsB:9-23 and GAD65) driving activation of CD4+ T cells in T1D patients. The goal of this study was to identify and optimize a monoclonal antibody that binds to DQ8 molecule and blocks the continued activation of DQ8 restricted T cells.

Methods: Hybridoma clones were generated from spleens of BALB/c mice immunized with DQ8 molecule bound with the InsB:9-23 peptide and screened by high throughput flow cytometry using human B-cells expressing DQ8. 28 clones found positive by flow cytometry were functionally tested in a mixed lymphocyte reaction containing a murine T cell line expressing a human TCR specific for the InsB: 9-23 - DQ8 complex. DQ8-103, a murine version of our best candidate antibody (DQ8-27), was tested in transgenic DQ8 mice immunized with GAD65 peptide; Cytokine production from the recall response ex vivo and in vivo was evaluated using a Luminex assay. Immunophenotyping of blood, spleens and draining lymph nodes isolated from immunized DQ8 mice was done by mass cytometry (CyTOF).

Results: We generated a monoclonal antibody against DQ8 (DQ8-27), that was able to bind human DQ8 molecule expressed on human B-cells and on murine splenocytes isolated from transgenic DQ8 mice. The murine version of DQ8-27 (DQ8-103) was able to significantly inhibit T cell activation in DQ8 mice immunized with GAD65 both ex vivo and in vivo, by decreasing the production of pro inflammatory cytokines IL-2 and IFN-γ. Of note, DQ8-103 caused a significant depletion of DQ8+ B-cells in treated mice compared to mice injected with a control antibody.

Disclosure

A. Lombardi: None. M. Zhu: None. T. Moran: None. T.A. Kraus: None. M. Hsieh: None. C. Stevens: None. D. Murphy: None. C.B. Donovan: Employee; Self; Pfizer Inc.. K. Atkuri: None. D.M. Messing: None. J. Chamoun: None. T.P. Brown: None. G. Guntas: None. D.V. Bennett: None. E. Concepcion: None. K. Johnson: None. Y. Tomer: None.
Associations between Executive Dysfunction and Disordered Eating Behavior (DEB) Risk in Teens with Type 1 Diabetes (T1D)
Authors: CECILIA-COSTA RAQUEL , HANSMANN MEREL , HARRINGTON KARA R. , VOLKENING LISA , LAFFEL LORI M.
Published: June 2018

Background: The adolescent brain remains immature with ongoing development in areas such as executive function (EF). DEBs also commonly emerge during adolescence. We explored associations between EF and risk of DEBs in teens, ages 13-17, with T1D.

Methods: EF was assessed by the Behavior Rating Inventory of Executive Function (BRIEF), both teen self-report and parent proxy-report versions. Scores ≥60, as total (Global Executive Composite [GEC]) score or on 2 indices (Behavioral Regulation [BRI] and Metacognition [MI]) or 8 subscales, indicate EF problems. DEB risk was assessed by the Diabetes Eating Problem Survey Revised (DEPS-R). DEB risk was categorized by DEPS-R scores: low risk <10; moderate risk 10-19; high risk ≥20.

Results: Teens (N=169, 46% female) were aged 15.9±1.3 (M±SD) years and had T1D for 8.4±3.7 years; A1c was 8.5±1.2% and 44% were overweight/obese. Based on teen self-reported DEPS-R score, 59% had low DEB risk, 29% moderate risk, and 12% high risk. Higher DEB risk was significantly associated with EF problems by teen self-report on GEC, BRI, and MI and by parent report on BRI (Figure). EF subscales associated with DEB risk included emotional control, organization, task completion, goal setting, and problem-solving.

Conclusion: In teens with T1D, executive dysfunction was associated with DEB risk, suggesting need for future study of the relationship between EF and DEBs.

Disclosure

R. Cecilia-Costa: None. M. Hansmann: None. K.R. Harrington: None. L. Volkening: None. L.M. Laffel: Consultant; Self; Eli Lilly and Company, Novo Nordisk Inc., Sanofi US, MannKind Corporation, Roche Diagnostics Corporation, Dexcom, Inc., Insulet Corporation, AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Johnson & Johnson Diabetes Institute, LLC..
The Metabolic Drivers of Neuropathy in India
Authors: CALLAGHAN BRIAN C. , VISWANATHAN VIJAY , REYNOLDS EVAN L. , FELDMAN EVA L.
Published: June 2018

Recent studies demonstrate that diabetes, prediabetes, and obesity are the main metabolic components associated with neuropathy. However, none of these studies were performed in India, where different waist circumference cutoffs exist. As part of an ongoing study, we are enrolling new patients from the M.V. Hospital for Diabetes in Chennai, India. Neuropathy was defined as a score of 2.5 or greater on the Michigan Neuropathy Screening Instrument (MNSI) Examination. Secondary neuropathy outcomes included the MNSI questionnaire, monofilament, biothesiometer, and sural device. Metabolic syndrome (MetS) components were defined using the International Diabetes Federation definition with South Asian waist circumference cutoffs. We determined the prevalence of neuropathy stratified by glycemic (ADA definition based on the glucose tolerance test). Multivariable logistic regression models were used to assess the association of metabolic components with neuropathy. To date, 264 patients have been recruited. Neuropathy was present in 8.7% of those with normoglycemia, 10.3% with prediabetes, and 23.7% with diabetes (p=<0.01) (Table). Diabetes (OR=3.87, 95% CI 1.05-14.28) was the only metabolic component associated with neuropathy. The prevalence of neuropathy increases with worsening glycemic status, but larger numbers are needed to determine which specific MetS components, other than diabetes, are the main drivers of neuropathy in India.Neuropathy status stratified by glycemic statusNeuropathy outcomeNormoglycemiaN (%)Pre-diabetesN (%)DiabetesN (%)MNSI exam4 (8.7%)9 (10.3%)31 (23.7%)MNSI questionnaire2 (4.4%)3 (3.5%)26 (20.3%)Monofilament0 (0.0%)3 (3.5%)6 (4.6%)Biothesiometer0 (0.0%)5 (6.1%)28 (22.6%)Sural device5 (17.9%)16 (23.5%)45 (43.3%)

Disclosure

B.C. Callaghan: Research Support; Self; Impeto Medical. Other Relationship; Self; American Academy of Neurology. Consultant; Self; Immune Tolerance Network. V. Viswanathan: None. E.L. Reynolds: None. E.L. Feldman: None.
HBsAg-1018, a Two-Dose Hepatitis B Vaccine, Is Well Tolerated and Effective in Diabetic Patients Aged 60 Years or Older
Authors: HYER RANDALL N. , JANSSEN ROBERT S.
Published: June 2018

Background: Adult patients with diabetes mellitus (DM) have a greater risk of contracting hepatitis B virus (HBV) than the general population, including those ≥ 60 years of age. When infected, patients with DM have more severe HBV-related morbidity and accelerated progression. The CDC recommends HBV vaccination of all adults with DM who are ≥ 60 years of age at the discretion of the treating physician. Due to persistent challenges, including hyporesponsiveness with current 3-dose vaccines, 87% of individuals with DM who are ≥ 60 years of age remain unvaccinated. HBsAg-1018 (Heplisav-BTM) is a 2-dose HBV vaccine with demonstrated higher protection rates than a 3-dose vaccine, particularly in populations known to be hyporesponsive. In this post-hoc analysis of a phase 3 trial, we assessed the safety and efficacy of HBsAg-1018 in patients with type 2 DM (T2DM) who were ≥ 60 years of age.

Methods: Patients with T2DM and who were 60-70 years old received either 2-dose HBsAg-1018 at 0 and 4 weeks and placebo at 24 weeks or 3-dose HBsAg-Eng (Engerix-B®) at 0, 4, and 24 weeks. Peak (week 24, HBsAg-1018 vs. week 28, HBsAg-Eng) seroprotection rates (SPRs) and safety were assessed by subgroups (sex, body mass index [BMI], and smoking status).

Results: A total of 480 patients (HBsAg-1018, n=327; HBsAg-Eng, n=153) were included in this analysis. Among patients who received all study injections, the peak SPR was significantly higher with HBsAg-1018 at week 24 (88.2% [231/262]) than with HBsAg-Eng at week 28, giving an overall difference of 29.7% (95% CI: 20.5, 39.1; P<.0001). SPRs with HBsAg-1018 were significantly higher compared with HBsAg-Eng regardless of sex, BMI, or smoking status (P ≤ .01). Adverse events, serious adverse events, and deaths were comparable between HBsAg-1018 and HBsAg-Eng.

Conclusion: Two-dose HBsAg-1018 provides better protection against HBV than a 3-dose vaccine (HBsAg-Eng) with a similar safety profile in patients ≥ 60 years of age with T2DM regardless of subgroup.

Disclosure

R.N. Hyer: Employee; Self; Dynavax Technologies. R.S. Janssen: Employee; Self; Dynavax Technologies.
High Level of Gingival Inflammation in Subjects with Type 1 Diabetes Despite Regular Dental Visits and Limited Microvascular Complications
Authors: ROY MARGAUX , GASTALDI GIACOMO , MOMBELI ANDREA , GIANNOPOULOU CATHERINE
Published: June 2018

The aim of the study was to evaluate small fiber neuropathy and periodontal health conditions on a cohort of patients with type 1 diabetes. The study participants were 18 females and 23 males with a mean age of 37.9 years (age range: 18-83 years). First, a routine medical examination was conducted at the Diabetology Clinics of the Geneva Universtity Hospital. Electrochemical skin conductance (ESC) was measured by Sudoscan, a noninvasive tool for detecting diabetic small fiber neuropathy. Participants were further examined for presence of periodontal disease by means of Plaque and Gingival indices (PI and GI respectively), probing depth (PD) and bleeding on probing. Self-reported attitudes and behavior regarding dental care was also collected during the visit. The mean duration of affliction with diabetes was 14.6 ±11.9 years, with a mean glycosylated heamoglobin (HbA1c) of 7.96±1.68%. The Sudoscan measurements showed an average ESC value between right and left sides for the hands of 68.2±13.4 microSiemenns (µS) and for the feet an ESC value of 79±6.9 µS. Seven percent of the subjects presented moderate dysfunction (ESC values 60-40 µS) whereas only one subject presented severe dysnfunction (< 40 µS). The periodontal examination revealed that all patients (except from 2) had signs of gingivitis, defined as presence of at least 2 sites with a GI>1. Gingival inflammation and bleeding on probing was present in 66% of the participants in more than 20% of sites independently of the presence of small fiber neuropathy. Periodontitis was observed in 15% of the subjets with a mean of 13 sites with PD>4mm and bleeding on probing. The majority of subjects (70%) reported having regular dental visits at least once per year. In the present cohort of patients with type 1 diabetes the main finding was the occurrence of pronounced gingival inflammation in the majority of subjects independently of small fiber neuropathy and despite regular dental visits.

Disclosure

M. Roy: None. G. Gastaldi: Speaker's Bureau; Self; Abbott, Eli Lilly and Company, Novo Nordisk A/S, Medtronic, Dexcom, Inc., Sanofi, Roche Diabetes Care Health and Digital Solutions, Boehringer Ingelheim GmbH. A. Mombeli: None. C. Giannopoulou: None.
The Impact of Diabetic Peripheral Neuropathy on Pinch Proprioception
Authors: YAHYA ABDALGHANI , KLUDING PATRICIA , PASNOOR MAMATHA , SANTOS MARCIO
Published: June 2018

Background: Deficits in finger proprioception might impact the ability to perform tasks such as picking up pills, monitoring blood sugar and injecting insulin. However, no tests are available for finger proprioception with functional relevance for tasks using pinch between the index finger and thumb. This study investigates the impact of diabetic peripheral neuropathy (DPN) on pinch proprioception in people with type 2 diabetes (T2D).

Methods: This study tested pinch proprioception in 8 healthy participants (3 males, 57±5.6 years), 11 participants with T2D without DPN (T2D-only) (5 males, 61±5.9 years), and 13 participants with T2D and DPN (T2D+DPN) (6 males, 59.9±6.4 years). We used a device previously tested for reliability. Following familiarization with a 15° target position of the index finger and thumb, subjects performed 3 trials where they attempted to reproduce the target position without visual feedback. The average of the absolute error matching the 15° target represented accuracy, and the standard deviation from the trials denoted precision.

Results: Descriptive statistics presented as Means ± SD. Accuracy: healthy (0.67±0.64), T2D-only (0.94±1.2), and T2D+DPN (3.51±2.24).

Precision: healthy (0.59±0.41), T2D-only (0.79±0.53), and T2D+DPN (2.81±2.4). Kruskal-Wallis test was significant for accuracy (p = .003) and precision (p = .001). Pinch proprioception was significantly different between the T2D+DPN group and the T2D-only group (accuracy and precision: p = .003) as well as between the T2D+DPN group and the healthy group (accuracy: p = .006; precision: p = .001). There were no significant differences between the T2D-only group and the healthy group: accuracy (p = .98) and precision (p = .5).

Conclusion: Our results showed that pinch proprioception was disrupted in people with DPN, but not in people with T2D without DPN. Detection of pinch proprioceptive deficits will potentially improve the neurological screening and can be used in future studies for other neurological populations.

Disclosure

A. Yahya: None. P. Kluding: None. M. Pasnoor: None. M. Santos: None.
Improving Management of Diabetic Dyslipidemia—Effect of Online CME
Authors: SPYROPOULOS JELENA , HEALY COLLEEN S.
Published: June 2018

Introduction: Although statins represent the cornerstone of dyslipidemia treatment, certain statins may increase the incidence of new-onset diabetes mellitus (DM). This study was conducted to determine if online continuing medical education (CME) could improve knowledge/competence of diabetologists/endocrinologists (D/E) and primary care physicians (PCPs) in tailoring statin therapy to reduce the risk for DM.

Methods: Physicians participated in at least one of two 30-minute video-based online CME activities. The activities addressed educational themes related to effects of statins on lipid metabolism and tailoring statin therapy to reduce the risk of DM. 6 matching pre-education/post-education questions associated with these themes were analyzed by comparing participant responses from pre- to post-education using a repeated-pairs design. For all questions combined, the McNemar’s chi-square test was used to assess differences from pre- to post-assessment.

Results: Significant improvements (P < .05) were seen after education across both CME activities for D/E (N=438) and PCPs (N=1023). For all questions combined, average relative improvements in knowledge/competence were 44% for D/E and 73% for PCPs (P<.05). Pre-assessment, the average correct response rate was 43% for D/E and 48% for PCPs, while post-assessment average correct response rates were 62% and 83%, respectively. Significant improvements were observed (all P<.05): effect of statins on glucose metabolism: 49% relative improvement among D/E (43% vs. 64%) and 103% improvement among PCPs (37% vs. 75%). Tailoring statin therapy to reduce the risk of DM: 64% improvement among D/E (50% vs. 82%).

Conclusions: This study demonstrates the success of online CME on improving knowledge/competence of D/E and PCPs regarding tailoring lipid-lowering strategies in high-risk patients. Properly implemented, this type of intervention can translate into improvements in guideline-based patient care and improve long-term outcomes for at-risk patients.

Disclosure

J. Spyropoulos: None. C.S. Healy: None.
Circulating PCSK9 in Statin-Treated Patients with Type 2 Diabetes Mellitus (T2DM)
Authors: WALUS-MIARKA MALGORZATA , KAPUSTA MARIA , KAWALEC EWA , IDZIOR-WALUS BARBARA
Published: June 2018

Proprotein Convertase Subtilisin/Kexin type 9 (PCSK9) is a natural circulating inhibitor of the LDL receptor. There is evidence that PCSK9 is a risk factor for coronary artery disease. In this study we aimed to determine factors associated with PCSK9 in patients with DMt2, according to statin treatment. Material included 116 consecutive patients with DMt2. Fasting serum lipids were determined enzymatically, apolipoproteins B100, A1 by immunoturbidimetry, apo C3, B48, pentraxin 3 (PTX3) and PCSK9 by ELISA, glycated hemoglobin A1c by HPLC. In the examined group 67 persons were statins users. Mean (SD) age of patients was 59.1 (11.1) years, HbA1c 8.6 (2.4%). Mean (SD) values of serum LDL-cholesterol were 2.6 (1.16) mmol/l and of serum PCSK9 255,4 (106,97) ng/ml. Patients treated with statins were older and had lower LDL-C: 2.38 (1.1) vs. 2.96 (1.2) mmol/l, p<0.05, apoB 100: 81.8 (20.6) vs. 99.7 (29.8) mg/dl, p<0.001 and higher waist circumference p<0,05. We did not observe differences in PCSK9 between patients receiving and not receiving statins: 248.6 (99.9) vs. 272.1 (112.5) ng/ml, p=0.2488 respectively. Median (IR) of apo B48 was lower in statin treated patients: 3529 (2370) vs. 5136.5 (3424.5) ng/dl, p<0.01. Correlation analysis revealed in both groups of patients significant correlation between PSCK9 and total cholesterol, LDL-C, triglycerides, PTX3 and apo B100. In statin users PCSK9 concentration was associated with apo A1 (p<0.001) and apo B48 (p<0.05). In patients not taking statins negative correlation between HbA1c and PCSK9 was found (p <0.05). The results indicate, that circulating PCSK9 is significantly associated with atherogenic markers - LDL-C, apo B 100, triglycerides and inflammation marker PTX3; in statin users also with apo A1, a negative risk marker, and apo B 48. In patients not taking statins significant negative association between PCSK9 and HbA1c was noted. These associations might be of value in understanding the pathways involved in atherosclerosis and dyslipidemia, mediated by PCSK9.

Disclosure

M. Walus-Miarka: None. M. Kapusta: None. E. Kawalec: None. B. Idzior-Walus: None.
Maternal High-Fat Diet Persistently Alters Bone Marrow Immune Cell Proportions and Function in a Nonhuman Primate Model
Authors: NASH MICHAEL J. , SODERBORG TAYLOR K. , JANSSEN RACHEL C. , PIETRAS ERIC M. , FRIEDMAN JACOB E.
Published: June 2018

Maternal obesity is rapidly increasing in prevalence and predisposes offspring to many serious health risks such as cardiovascular disease, obesity, and nonalcoholic fatty liver disease. Maternal high-fat diet (MHFD) can alter offspring metabolism and predispose offspring to obesity through many mechanisms, including alterations in immune cell function. However, whether maternal diet impacts development of the neonatal immune system remains unclear. To investigate the impact of MHFD on offspring bone marrow, a major site of immune cell generation via hematopoiesis, we utilized two distinct cohorts of non-human primates (NHPs). Bone marrow from a cohort of early third trimester fetuses was examined to determine the effects of MHFD vs. chow diet on bone marrow cellularity and function. The second cohort, 3-year-old juvenile NHPs exposed to MHFD early in life, then shifted to a chow diet at weaning for the remaining 2.5 years, were studied to see whether bone marrow phenotypes resulting from MHFD exposure were reversible upon altering diet later in life. Colony-forming assays of plated fetal bone marrow cells showed a significant 34.5% relative increase in innate immune system cells at the expense of erythroid (-78.9%) and multilineage (-53.8%) progenitors, and a decrease in total numbers of all cell types. Bone marrow stem cells differentiated to macrophages were treated with LPS and showed significantly lower IL1B expression and trending decreased IL10 and TNF via qPCR, suggesting hyporesponsivity to inflammatory LPS stimuli. Similar phenotypes were found in the cohort of juveniles exposed to MHFD then shifted onto a chow diet, suggesting that exposure to MHFD has long lasting immunological consequences in offspring not correctable by dietary changes later in life. Further, our results suggest remodeling in macrophage function, potentially implicating an epigenetic programming event early in life driven by maternal diet.

Disclosure

M.J. Nash: None. T.K. Soderborg: None. R.C. Janssen: None. E.M. Pietras: None. J.E. Friedman: Advisory Panel; Self; Janssen Pharmaceuticals, Inc..
Combination of Premixed Insulin with Glucagon-Like Peptide-1 Receptor Agonist (GLP-1) for Uncontrolled Type 2 Diabetes Mellitus (T2DM) Patients
Authors: FAZELI SASAN , EHRHARDT NICOLE
Published: June 2018

Background: Despite multiple therapeutic options for people with T2DM, 48% of people do not achieve Hemoglobin A1c (HbA1c) goal. GLP-1 receptor agonists may be beneficial to patients with T2DM who are on large insulin doses and are obese. GLP-1 receptor agonists are often used with oral medications and/or basal insulin. GLP-1 agents are not typically used with those on prandial insulin.

Clinical cases: We present 2 cases of uncontrolled T2DM whose HbA1c significantly improved on GLP-1 and daily mixed insulin. Patient A is a 62 y/o M with T2DM for 15 years, weight of 112 kg (BMI 35), HbA1c of 10.2% (diabetes ref range > 6.4%), on 65u insulin detemir BID (total daily insulin 0.9 u/kg). HbA1c decreased to 7.1% by 2 month on dulaglutide 1.5mg weekly, insulin Humalog mix 75/25 45u BID (total daily insulin 0.8u/kg) and metformin ER 1000mg BID. HbA1c at 12 months was 6.1% and weight 112kg (BMI 35). Patient B is a 61 y/o F with T2DM for 17 years, weight of 92 kg (BMI 35), HbA1c of 10.9%, on 50u insulin glargine BID and not adherent to her insulin lispro (total daily insulin 1.6 u/kg). HbA1c decreased to 6.3% at 2 month on dulaglutide 1.5 mg weekly, insulin Novolog mix 70/30 30u BID (total daily insulin 0.7u/kg) and metformin ER 500mg BID. HbA1c at 12 months was 6.6%, and weight decreased to 87kg (BMI 34).

Discussion: Our patients were on metformin and high dose insulin without HbA1c improvement. Their regimens were changed to once weekly GLP-1 agonists with mixed insulin which showed improved and sustained glycemic control after one year without major hypoglycemic events.

Conclusion: These cases highlight the potential utility of GLP-1 receptor agonist therapy in combination with mixed insulin for patients with uncontrolled T2DM on high dose insulin. We hypothesize that increased compliance may be an import factor but further research needs to be completed.

Disclosure

S. Fazeli: None. N. Ehrhardt: None.