bluuebunny/crossref_metadata_2025 huggingface dataset:
2080-P: Lipid Profiles from Individual Cells of the Human Pancreatic Islet
Authors: RUBAKHIN STANISLAV , SWEEDLER JONATHAN V.
Published: June 2020

Lipids play multiple roles in normal and pathological functions of cells and organs including structural, energy balance, cell signaling and maintenance of homeostasis. In the pancreatic islets, lipids have been implicated in regulating aspects of membrane fusion and hence insulin (and other hormones) release. Imbalances in lipid metabolism, accumulation, and localization have been associated with T2D progression. Wide variety of lipid classes are observed in the pancreatic islets including glycerophospholipids, glycerophospholipids, and ceramides. Complex cellular organization of these structures suggests that lipid profiles of different cell types or even functional cell subtypes, may not be the same. In this work, high throughput single cell mass spectrometry (MS) was used to analyze cellular populations of isolated human individual pancreatic islets received from the Human Pancreas Analysis Program (UPenn). Besides lipids, peptide markers for the canonical islet cell types are observed in the same cells allowing us to associate specific lipid profiles with cell types. For example, beta cells exhibited larger signals matching to PC(34:1), SM(d34:2), and SM(d38:1). Epsilon cells, among others, have enriched SM(d42:2)- and SM(d42:1)-like signals. Alpha cells demonstrate the most complex lipid profile with 10 lipid species exhibiting higher relative intensities. Current work involves matching the single cell lipid peaks to specific lipids, and understanding how the high plasticity of lipid profiles in each cell type track disease progression. These observations create a base for further investigation of roles of lipids in normal and pathological functioning of the pancreatic islets.

Disclosure

S. Rubakhin: None. J.V. Sweedler: None.

Funding

American Diabetes Association/Pathway to Stop Diabetes (1-18-VSN-19 to J.V.S.)
2086-P: Peptidyl Hormone Detection in Formaldehyde-Fixed Pancreas Using Mass Spectrometry Imaging
Authors: LEE DONGKYU , RUBAKHIN STANISLAV , SWEEDLER JONATHAN V.
Published: June 2020

The availability of well-documented high-quality clinical pancreas specimens in tissue depositories such as the nPOD allows multiplatform comparative investigations of the biochemical correlates of T1D and T2D disease progression. Mass spectrometry imaging (MSI) is an emerging technique that allows us to determine, in an untargeted manner, the proteins and peptides and monitor their localization within the pancreas. However, its application to specimens preserved using standard formaldehyde-induced crosslinking is limited to molecules that do not have primary amines, and thus excludes endogenous peptides and proteins. Here we are creating a MSI-compatible chemical decrosslinking to enable us to use the tissue banked materials. We found that hydroxylamine hydrochloride in acidic tris(hydroxymethyl)aminomethane reverses the crosslinking between model peptides in vitro. Among amino acid moieties affected by formaldehyde-fixation, arginine, lysine and cysteine demonstrate best performance leading to the restoration of peptide detection in the treated samples. This treatment is compatible with formaldehyde-fixed pancreas and successfully enables the detection of glucagon, insulin and pancreatic polypeptides localized in the α-, β- and γ-cells. Our approach opens important opportunities for multimodal investigation of large sets of well preserved and documented clinical specimens.

Disclosure

D. Lee: None. S. Rubakhin: None. J.V. Sweedler: None.

Funding

American Diabetes Association/Pathway to Stop Diabetes (1-18-VSN-19 to J.V.S.)
209-OR: Evidence that Agrp Neuron Activation Drives Cold-Induced Hyperphagia
Authors: DEEM JENNIFER D. , FABER CHELSEA L. , PEDERSEN CHRISTIAN , PHAN BAO ANH N. , OGIMOTO KAYOKO , LARSEN SARAH A. , TRAN MEGAN A. , DAMIAN VINCENT , KAIYALA KARL , SCARLETT JARRAD , BRUCHAS MICHAEL R. , SCHWARTZ MICHAEL W. , MORTON GREGORY J.
Published: June 2020

Although hyperphagic feeding during cold exposure is widely viewed as a response to the negative energy state that results from increased thermogenesis, the underlying mechanisms are unknown. We investigated an alternative hypothesis, that during cold exposure, hyperphagia is mounted in anticipation of homeostatic need, rather than as a compensatory response. In support of this hypothesis, we report that in chow-fed mice acutely housed at 14°C, increases of food intake and thermogenesis occur both rapidly and simultaneously, implying hyperphagia is not mounted as a compensatory response. We next asked whether this cold-induced feeding response involves activation of agouti-related peptide (Agrp) neurons, which are implicated in other forms of hyperphagic feeding. Consistent with this hypothesis, we report that Agrp neurons are activated within 1-2 min of cold exposure and prior to the onset of hyperphagia, as judged by in vivo fiber photometry and confirmed by cFos induction (Agrp+/cFos+, 22°C: 5.3±4.5% vs. 14°C: 20.2±6.1%; p<0.01). To test whether Agrp neuron activation is required for cold-induced hyperphagia, we employed a chemogenetic strategy to silence Agrp neurons. As predicted, their silencing prevented hyperphagic feeding (2h food intake after housing at 14°C: 0.26±0.06 g vs. 0.10±0.04 g; p<0.05), but not the associated thermogenic response (0.63 vs. 0.62 kcal/hr; p=ns) during cold exposure. Interestingly, cold-induced hyperphagia is absent in mice with diet-induced obesity (DIO). Consequently, unlike chow-fed mice, these animals lose weight during cold exposure. These observations suggest that 1) Agrp neurons lie downstream of thermoregulatory neurocircuits, 2) during cold exposure, increased Agrp neuronal firing occurs rapidly as part of a homeostatic response that preserves body temperature and energy stores, and 3) this response appears to be impaired in mice with DIO. Future studies will map the neurocircuitry underlying these responses and clarify how DIO impairs their activation.

Disclosure

J.D. Deem: None. C.L. Faber: None. C. Pedersen: None. B.N. Phan: None. K. Ogimoto: None. S.A. Larsen: None. M.A. Tran: None. V. Damian: None. K. Kaiyala: None. J. Scarlett: None. M.R. Bruchas: None. M.W. Schwartz: Research Support; Self; Novo Nordisk A/S. G.J. Morton: Research Support; Self; Novo Nordisk A/S.

Funding

American Diabetes Association (1-19-PDF-103 to J.D.D.); Dick and Julia McAbee Foundation; National Institutes of Health (T32-HL007028-39, R01DK089056, R01DK101997, R01DK124238, R01DK089056, P30DK017047, P30DK035816)
2091-P: Effects of Caveolin-1 on the Synthesis and Decomposition of Islet ß-Cell IAPP and Its Mechanism
Authors: LIU KUNYING , ZENG WEN , LIN SHUO , LIN CHUWEN , XU FEN , CAI NAN , ZENG LONGYI
Published: June 2020

Objective: Islet amyloid polypeptide(IAPP) is a major component of islet amyloid deposition in type 2 diabetic patients. Our preliminary study showed that membrane protein Caveolin-1 (Cav1) silencing inhibited β-cell apoptosis and promote insulin secretion. The purpose of this study was to investigate the regulation of Cav1 on islet β-cell IAPP and its mechanism.

Materials and Methods: Firstly, IAPP secretion of each group of primary islets was detected by ELISA, then Western blot and real-time quantitative PCR (qPCR) were used to detect the expression levels of IAPP, PAM(Peptidyl-glycine alpha-amidating monooxygenase), PC1(Prohormone Convertase-1), BACE2(Beta-secretase 2), IDE(Insulin-Degrading Enzyme), TXNIP(Thioredoxin interacting protein), and finally, it confirmed that Cav1 interacted with the protein of TXNIP by immunofluorescence confocal and co-immunoprecipitation (Co-IP).

Results: The secretion of IAPP was decreased in primary pancreatic islets and Cav1 deficiency significantly down-regulated the expression of IAPP in NIT-1 cells, while over-expression of Cav1 in βTC-6 cells significantly up-regulated the expression of IAPP. This effect correlated with Cav1 silencing leading to down-regulation of PAM expression associated with IAPP synthesis and over-expression of Cav1 leading to up-regulation of PC1 associated with IAPP synthesis. On the other hand,Cav1 deficiency resulted in the up-regulation of the expression of BACE2 and IDE, the enzymes associated with IAPP decomposition. While over-expression of Cav1 leaded to down-regulation of BACE2 and IDE. Further studies indicated that Cav1 co-localized with TXNIP, and it interacted with and regulated the expression of TXNIP, which regulates IAPP gene transcription.

Conclusion: Cav-1 can regulate the synthesis and decomposition of IAPP in islet β cell by interacting with TXNIP. This study suggests that Cav-1 may protect diabetic islet beta cells by reducing islet amyloid deposition.

Disclosure

K. Liu: None. W. Zeng: None. S. Lin: None. C. Lin: None. F. Xu: None. N. Cai: None. L. Zeng: None.

Funding

National Natural Science Funding of China (51873071); Natural Science Foundation of Guangdong Province (2018B030311012)
2103-P: Glucose-Dependent Insulinotropic Polypeptide (GIP) Protects against Cytokine-Induced Cell Death and Exerts Both Insulinotropic and Glucagonotropic Effects in Human Islets
Authors: JØRGENSEN ANNE , EGESKOV MERETE , KNOP FILIP K. , STØRLING JOACHIM
Published: June 2020

The incretin hormone glucose-dependent insulinotropic polypeptide (GIP) is secreted from enteroendocrine K cells following food intake and potentiates glucose-stimulated insulin secretion from β-cells. Recent studies suggest that GIP also has glucagonotropic properties under euglycemic and hypoglycemic conditions in humans. GIP may therefore be exploited as a safeguard against hypoglycemia in patients with type 1 diabetes (T1D). However, a direct glucagonotropic effect of GIP on human α-cells has not yet been investigated. In this study, we investigated the effects of GIP on glucagon and insulin secretion in isolated human islets during 30-min exposures to low (2 mM) and high (20 mM) D-glucose concentrations. We further investigated the potency of GIP to protect human islets from cell death induced by a 48-h exposure to pro-inflammatory cytokines (50 U/mL interleukin-1β + 1,000 U/mL interferon-γ) known to contribute to islet destruction in T1D. GIP (2.5 nM) significantly (P=0.048, n=4) increased glucagon secretion by 48% at 2 mM glucose in isolated human islets as compared to 2 mM glucose alone. At 20 mM glucose, GIP did not affect glucagon secretion, but potentiated insulin secretion by 45% (P=0.015, n=4). Further, cytokine-induced human islet cell death was decreased by 50% by co-treatment with GIP (P=0.001, n=3). Our findings demonstrate that GIP protects against cytokine-induced islet death and possesses glucagonotropic and insulinotropic effects on human islets at low and high glucose, respectively. These findings support GIP as a bifunctional blood glucose-stabilizer that might be exploited to treat T1D.

Disclosure

A. Jørgensen: None. M. Egeskov: None. F.K. Knop: Advisory Panel; Self; AstraZeneca, Merck Sharp & Dohme Corp., Mundipharma International, Novo Nordisk A/S, Sanofi. Consultant; Self; Carmot Therapeutics, Inc., Eli Lilly and Company, Novo Nordisk A/S. Research Support; Self; AstraZeneca, Gubra, Novo Nordisk A/S, Sanofi, Zealand Pharma A/S. Speaker’s Bureau; Self; AstraZeneca, Lupin Pharmaceuticals, Inc., Merck Sharp & Dohme Corp., Norgine B.V., Novo Nordisk A/S. J. Størling: None.
2118-P: Regulation of Ataxia-Telangiectasia and Rad3-Related (ATR)–Dependent DNA Damage Response by Nitric Oxide in ß-Cells
Authors: YEO CHAY TENG , OLESON BRYNDON , CORBETT JOHN A. , SCHNUCK JAMIE K.
Published: June 2020

Nitric oxide, generated in β-cells in response to cytokines, induces DNA damage that activates ataxia-telangiectasia mutated protein (ATM)-dependent DNA damage response (DDR). While the DDR does not contribute to DNA repair in β-cells, it does regulate β-cell apoptosis when the nitric oxide production falls below a threshold level. Indeed, nitric oxide at high micromolar levels, while inducing DNA damage, also inhibits DDR and protects β-cells from apoptosis. This ability of nitric oxide, a potent inhibitor of mitochondrial oxidation, to attenuate DDR activation correlates with a 10-fold reduction in ATP and the actions are selective to β-cells. Unlike most other cell types, β-cells cannot compensate for mitochondrial inhibition by increasing glycolysis, leading to a loss in ATP levels which correlates with DDR inhibition. We now show that nitric oxide also inhibits the activation of a second DDR transducer kinase, ATR. ATR is activated in response to single-strand DNA breaks and replication stress. We showed that low levels of nitric oxide activate ATR-dependent DDR signaling by inducing replication stress. However, much like with ATM, at higher concentrations nitric oxide and other inhibitors of mitochondrial respiration that reduce ATP levels by more than 10-fold, also inhibit ATR selectively in β-cells. Nitric oxide and these mitochondrial poisons do not inhibit ATR signaling in non-β-cells as they maintain ATP levels by increasing glycolysis under mitochondrial inhibition. Importantly, when non-β-cells are forced to utilize oxidative phosphorylation for ATP generation, nitric oxide and other mitochondrial inhibitors attenuate ATR signaling in a manner similar to β-cells. Together, these studies suggest that inhibition of the DDR by nitric oxide in β-cells is dependent on its ability to inhibit mitochondrial oxidative metabolism and deplete ATP, and suggest that the inhibition of DDR signaling may be a protective response to prevent β-cell apoptosis.

Disclosure

C. Yeo: None. B. Oleson: None. J.A. Corbett: None. J.K. Schnuck: None.

Funding

National Institutes of Health (DK052194, AI-44458)
2124-P: Raptor Levels Are Critical in the Adaptation of Beta Cells to High-Fat Diet
Authors: BLANDINO-ROSANO MANUEL , BERNAL-MIZRACHI ERNESTO
Published: June 2020

mTORC1 signaling is a central regulator of autophagy and altered autophagic activity has been implicated in the beta-cells of patients with type 2 diabetes, and in the beta-cells of obese mice. The importance of mTORC1 signaling and Raptor in beta-cells has been recently demonstrated using raptor knock-out models (βraKO mice). These studies have shown that complete deletion of Raptor impairs beta-cell proliferation and survival of the beta-cell, and both in function and insulin processing. However, nothing is known whether reduced mTORC1 levels impair beta-cell adaptation to diabetogenic conditions. Therefore, we decided to study mice with haploinsufficiency of Raptor in beta-cells (βraHET). Mice with heterozygous deletion of Raptor in beta-cells were generated by crossing raptorf/f with Rip-Cre mice (βraHET). βraHET and control mice were fed regular chow (RC) or high-fat diet (HFD). Mice fed with RC showed no difference at the metabolic level, islets morphology, or beta-cell function. βraHET mice fed HFD for eight weeks showed an impaired Intraperitoneal Glucose Tolerance Test, fed insulin secretion and a decreased beta-cell mass, compared to control mice. Isolated islets from βraHET mice fed HFD for eight weeks showed impaired insulin secretion either measured by perifusion or static incubation compared to control islets. Next, we tested whether isolated islets from βraHET mice are more susceptible to beta-cell stressors. Beta-cell death determined by TUNEL and cleavage Caspase 3 was increased (50 and 60%) in the βraHET islets after 24 h exposure to palmitate. In addition, autophagy markers such as LC3 and p62 were increased in islets from βraHET treated with palmitate. The increase in cell death and autophagy is specific to lipotoxicity since there βraHET islets exhibited no increase in cleavage Caspase 3 in TUNEL when they were treated with proinflammatory cytokines or ER Stress inducers.

In conclusion, we demonstrate that adequate levels of Raptor are critical in the adaptation of the beta-cell to HFD.

Disclosure

M. Blandino-Rosano: None. E. Bernal-Mizrachi: None.
2132-P: Single-Cell RNA-Sequencing of Mouse Islets after Inflammatory Cytokine Exposure
Authors: STANCILL JENNIFER , KHATUN ACHIA , KASMANI MOUJTABA , CUI WEIGUO , CORBETT JOHN A.
Published: June 2020

Type 1 diabetes is an autoimmune disease caused by destruction of pancreatic β-cells. During disease progression, β-cells are exposed to proinflammatory cytokines like interleukin-1β (IL-1β) and interferon-γ (IFN-γ), stimulating them to express inducible nitric oxide synthase (iNOS) and produce micromolar levels of nitric oxide. While some cytokine-mediated changes in islet gene expression are known, we do not have a global view of the genes induced by cytokines nor do we know how non-β-cells respond to cytokines. Following 6-hr treatment with IL-1β and IFN-γ, adult C57BL/6 mouse islets were subjected to single-cell RNA-sequencing using the 10x Genomics Chromium platform. iNOS mRNA (Nos2) was induced in 73% of β-cells in response to both cytokines. Unexpectedly, 72% of Δ-cells and 44% of α-cells expressed Nos2 at levels similar to that found in β-cells. This is surprising since β-cells are thought to be the primary islet source of nitric oxide. Of the 241 genes enriched in Nos2-expressing β-cells, 70% were also enriched in Nos2-expressing Δ- and α-cells, suggesting that these cells initially respond to cytokine exposure by activating similar pathways. Among the enriched genes were those indicative of anti-viral defenses, suggesting induction of protective responses. Nos2 expression was negatively correlated with expression of genes encoding heat shock proteins, such as Hsp70, consistent with previous reports that heat shock inhibits cytokine-induced iNOS expression. Finally, we observed IL-1β-induced repression of β-cell identity genes, including Mafa, Pdx1, Slc2a2, and Ucn3. Repression of these identity genes was negatively correlated with Hsp70 expression, suggesting that cellular stress prevents this dedifferentiation. While these studies are ongoing, they suggest that (1) Δ- and α-cells respond to cytokines by upregulating Nos2 mRNA, (2) cytokines repress β-cell identity genes, and (3) both of these outcomes can be inhibited by induction of cellular stress.

Disclosure

J. Stancill: None. A. Khatun: None. M. Kasmani: None. W. Cui: None. J.A. Corbett: None.
2139-PUB: Antihypertensive Effect of Azilsartan in Newly Diagnosed Stage 2 Diabetic Hypertensives
Authors: MAHAPATRA HARIBALLAV , KHUNTIA MONALISA , MISHRA SUSANT , MISHRA SANAT K. , PADHI RAJESH K. , JENA BISWARANJAN , DAS SREEYA , KHATUA RANJIT K. , JENA APURBA R. , MAHAPATRA RAMABALLAV , MAHAPATRA LAXMINARAYAN , SAHOO ABHAY K.
Published: June 2020

Background and Aim: Coexistence of hypertension (HTN) and type 2 diabetes (T2DM) multiplies the risk of cardiovascular events. Early identification and prompt management of elevated blood pressure in T2DM has shown to improve the quality of life and to reduce the economic burden to the patients. We studied blood pressure (BP) lowering effect of Azilsartan (AZST) monotherapy in newly diagnosed stage 2 hypertensives with a history of T2DM. We also analysed its effects on blood glucose and renal indices.

Methods: T2DM Subjects attending a specialized diabetes clinic, who were diagnosed with HTN for the first time, were invited to participate. Mean of 3 BP readings was considered for analysis after due consent. Subjects with stage 2 hypertension were administered AZST 40mg once daily. After 3 months they had a repeat BP recording. Subjects underwent serum creatinine, Na+, K+, fasting (FPG) and postprandrial (PPG) plasma glucose measurements at baseline and after 3moths’ of enrolment. We compared BP lowering efficacy and changes in plasma glucose and renal parameters at baseline and after 3months of AZST therapy. Diabetic, dyslipidemic and other chronic medicines were continued unchanged during the study period.

Results: Inclusion criteria were met by 474 subjects (117, 25% females). Mean age and duration of diabetes were 53.3 (95% CI 52.3-54.3) and 7.03 (CI 6.6-7.5) years respectively. With AZST therapy mean SBP dropped from 152.7 (CI 151.1-154.3) mm Hg to 136.4 (CI 134.9-138.0) mm Hg (p <0.05) and the post treatment DBP declined to 79.6 (CI 78.8-80.4) mm Hg from its baseline value of 90.5 (CI 89.6-91.4) mm Hg (p <0.05). The mean baseline FPG and PPG were 158.8 (CI 154.0-163.4) mg/dL and 237.3 (CI 231.9-242.7) mg/dL in order. There was a non-significant decline in plasma glucose, but no changes were seen in eGFR or electrolytes. No adverse event was reported.

Conclusion: Azilsartan monotherapy was found to be safe and effective in managing newly diagnosed stage 2 hypertension in type 2 diabetes without hampering renal function.

Disclosure

H. Mahapatra: None. M. Khuntia: None. S. Mishra: None. S.K. Mishra: None. R.K. Padhi: None. B. Jena: None. S. Das: None. R.K. Khatua: None. A.R. Jena: None. R. Mahapatra: None. L. Mahapatra: None. A.K. Sahoo: None.

Funding

Jyoti Diabetes Research Foundation
214-OR: Role of Hypothalamic MAPK/ERK Signaling in Diabetes Remission Induced by the Central Action of Fibroblast Growth Factor 1 (FGF1)
Authors: BROWN JENNY M. , PHAN BAO ANH N. , WASANWALA HUZAIFA , MATSEN MILES E. , SECHER ANNA , MORTON GREGORY J. , SCHWARTZ MICHAEL W. , SCARLETT JARRAD
Published: June 2020

The hypothalamic arcuate nucleus-median eminence (ARC-ME) is a target for sustained diabetes remission induced by fibroblast growth factor 1 (FGF1). To identify intracellular signaling cascades underlying this effect, we considered evidence that sustained signal transduction responses engaged by FGF1 are integrin-dependent. To determine if the sustained antidiabetic response of diabetic ob/ob mice to intracerebroventricular (icv) injection of FGF1 is also integrin-dependent, we investigated the response to a mutant FGF1 that is unable to activate integrin receptors (R50E FGF1). We found that although icv injection of R50E FGF1 mimics the transient anorexia, weight loss and blood glucose lowering responses induced by native FGF1 (P<0.001 vs. Veh for each), it fails to elicit sustained diabetes remission. Similarly, MAPK/ERK signaling is induced in the ARC-ME (primarily in tanycytes) for up to 24 h following a single icv FGF1 injection in both wild type and ob/ob mice, but the duration of this response is reduced by∼70% following icv R50E FGF1 injection (P<0.0001 vs. icv FGF1). We therefore hypothesized that owing to its integrin-dependent nature, FGF1-induced remission of diabetic hyperglycemia requires sustained activation of the MAPK/ERK signaling pathway in the ARC-ME. To test this hypothesis, 4 groups of diabetic ob/ob mice were studied. Each group received a single icv injection of either saline vehicle or recombinant FGF1 (3 µg), followed immediately by continuous icv infusion of either the MAPK inhibitor U0126 or its vehicle (DMSO) for 24 h. We report that the duration of the antidiabetic effect induced by icv FGF1 injection is attenuated by pharmacologic blockade of MAPK/ERK signaling. We conclude that the sustained remission of diabetic hyperglycemia induced by the central action of FGF1 depends upon its ability to activate hypothalamic MAPK/ERK signaling in a sustained manner.

Disclosure

J.M. Brown: None. B.N. Phan: None. H. Wasanwala: None. M.E. Matsen: None. A. Secher: Employee; Spouse/Partner; Gubra. Employee; Self; Novo Nordisk A/S. Stock/Shareholder; Self; Novo Nordisk A/S. G.J. Morton: Research Support; Self; Novo Nordisk A/S. M.W. Schwartz: Research Support; Self; Novo Nordisk A/S. J. Scarlett: None.

Funding

National Institutes of Health (K08DK114474, DK101997, DK083042, DK035816, P30DK017047, HL007312); University of Washington
2150-PUB: Chemotactic Cytokine Receptor 5 (CCR5) Gene Promoter Polymorphism (rs1799987) Is Associated with the Risk of Diabetic Kidney Disease in Polish Patients with Type 2 Diabetes
Authors: NABRDALIK KATARZYNA , POKRZYWNICKA PATRYCJA S. , KWIENDACZ HANNA , ADAMCZYK PIOTR , MOCZULSKI DARIUSZ , MOCZULSKA HANNA , DROZDZ KAROLINA , GRZESZCZAK WLADYSLAW , GUMPRECHT JANUSZ
Published: June 2020

Background and Aims: The environmental risk factors of diabetic kidney disease (DKD) do not allow identification of people at higher risk of it. A lot of expectations are being placed to the genetic predisposition. Among patients with diabetes, the presence of CCR5 gene polymorphism is suspected to be associated with DKD, yet the results are inconclusive. The aim of this study was to evaluate its association with DKD among Polish patients with type 2 (T2DM).

Materials and Methods: CCR5 gene polymorphism (rs1799987, 59029 A/G) was assessed among 467 consecutive patients with T2DM visiting one outpatient diabetology clinic in Upper Silesia, Poland. DKD diagnosis was based on urinary albumin creatinine ratio >300 mg/g and the presence of diabetic retinopathy. Information related to time of diabetes duration and HbA1c were collected.

Results: Clinical characteristic of studied patients is presented in Table 1. We found that both GG genotype (p=0.035) and allele G (p=0.02) were significantly associated with DKD occurrence and G allele was present in all of the DKD patients.

Conclusion: A significant association between rs1799987 polymorphism of CCR5 gene and the presence of DKD in the group of Polish patients with T2DM was found what is in accordance with some previous studies. Further studies are needed in this area to confirm this association.

Disclosure

K. Nabrdalik: None. P.S. Pokrzywnicka: None. H. Kwiendacz: None. P. Adamczyk: None. D. Moczulski: None. H. Moczulska: None. K. Drozdz: None. W. Grzeszczak: None. J. Gumprecht: Consultant; Self; Astra, Bioton, Boehringer Ingelheim International GmbH, Eli Lilly and Company, Merck & Co., Inc., Merck Sharp & Dohme Corp., Mundipharma International, Novo Nordisk A/S, Polfa Tarchomin S.A., Sanofi, Servier. Speaker’s Bureau; Self; Astra, Bioton, Boehringer Ingelheim International GmbH, Eli Lilly and Company, Merck & Co., Inc., Merck Sharp & Dohme Corp., Mundipharma International, Novo Nordisk A/S, Polfa Tarchomin S.A., Sanofi, Servier.
2157-PUB: The Effects of an External Shoe-Lift on Off-Loading the Diabetic Foot
Authors: ERSEN ALI , JUPITER DANIEL , RASPOVIC KATHERINE M. , LAVERY LAWRENCE A. , YAVUZ METIN
Published: June 2020

Diabetic foot ulcers (DFUs) are a complication of diabetes, reducing patients’ quality of life, increasing morbidities, and contributing to more the $30B annually in increased for the U.S. healthcare system. Total Contact Cast (TCC) is considered the gold standard for healing DFUs. Within the last decade a number of prefabricated TCC kits such as TCC-EZ have been introduced, which offer a user-friendly casting process for healthcare providers. However, these casts and their associated boots or shoes cause asymmetry in gait. They may also result in reduction of weight bearing on the casted side and eventually affect the healing of DFUs. The purpose of this exploratory study was to determine the effectiveness of a shoe balancer called Evenup in redistribution of plantar peak pressure (PP) during gait. Sixteen subjects provided informed consent, 5 of whom had diabetic neuropathy (one Type I, four Type II), and completed three trials of 30-m walking while fitted with TCC-EZ and TCC, either with or without Evenup, in a randomized order. A pair of automated wireless photogate sensors captured time to traverse the distance, and pedobarographic insoles (Novel, Germany) measured and recorded plantar pressures. For each trial, the mid-gait six steps for each side were analyzed. Paired t-tests were used to compare bilateral PP difference, as well as whether presence of Evenup changed differences in pressure side to side. Bilateral PP difference in TCC (p=0.166) and TCC-EZ (p=0.123) arms did not reveal significance due to presence of Evenup (p>0.05) while both modalities showed significant PP reduction side to side (p≤0.001). More research is necessary to reveal the potential clinical benefits of the use of shoe lifts in diabetic foot offloading, falls in diabetic patients and healing rates in a larger cohort.

Disclosure

A. Ersen: None. D. Jupiter: None. K.M. Raspovic: Consultant; Self; Orthofix. L.A. Lavery: Advisory Panel; Self; Acelity, Bayer AG. Consultant; Self; Boehringer Ingelheim International GmbH. Research Support; Self; AstraZeneca, Cardinal Health, EO2 Concepts, Integra LifeSciences, MedImmune, Smith & Nephew. M. Yavuz: Research Support; Self; Integra LifeSciences. Other Relationship; Self; Vivonics Inc.

Funding

Integra LifeSciences
218-OR: Does Antibiotic Treatment before Bone Biopsy Affect the Identification of Bacterial Pathogens from Bone Culture?
Authors: LAVERY LAWRENCE A. , RYAN EASTON C. , TRUONG DAVID H. , JOHNSON MATTHEW J. , FONTAINE JAVIER LA , CRISOLOGO PETER A.
Published: June 2020

Our aim was to evaluate the effect of antibiotic exposure on bacterial pathogen yield in patients with diabetic foot osteomyelitis. In a retrospective cohort study, we evaluated 142 patients admitted to the hospital with diabetic foot osteomyelitis which confirmed by histologic evaluation. Histologic criteria for osteomyelitis included the presence of an inflammatory response, bone necrosis, and/or bone fragmentation. We compared bacterial pathogens in patients that received antibiotics before bone culture (n=114) and patients that did not receive antibiotics (n=28), as well as bacterial yield based on the duration of antibiotic exposure. We used the chi square test to compare antibiotic exposure and culture yield. There was not a significant difference in culture yield in patients that received antibiotics (77.9%) and patients that did not (85.7%, p=0.58). There was no difference in patients that received antibiotics for ≥4 days (n=56, 71.4%), 1-3 days (n=58, 84.5%) and no antibiotics (85.7%, p=0.15).

In conclusion, the results of bacterial recovery were similar in patients regardless of the length of antibiotic treatment. Antibiotic exposure did not change the bacterial yield, the type of bacteria or the number of bacteria. In patients with suspected diabetic foot osteomyelitis, there is no evidence to support withholding antibiotics prior to obtaining a bone biopsy.

Disclosure

L.A. Lavery: Advisory Panel; Self; Acelity, Bayer AG. Consultant; Self; Boehringer Ingelheim International GmbH. Research Support; Self; AstraZeneca, Cardinal Health, EO2 Concepts, Integra LifeSciences, MedImmune, Smith & Nephew. E.C. Ryan: None. D.H. Truong: None. M.J. Johnson: None. J. La Fontaine: None. P.A. Crisologo: None.
2202-PUB: Do Women with Diabetes in Pregnancy Have Better Medication Adherence to Long-Acting Insulin Compared with Intermediate Insulin?
Authors: NAZEER SARAH A. , HERRICK CYNTHIA J. , DUCKHAM HILLARY E. , CARTER EBONY B.
Published: June 2020

Objective: To determine whether women with pre-existing and gestational diabetes have better medication adherence and pregnancy outcomes when treated with long acting, compared to intermediate acting, insulin.

Methods: We performed a retrospective cohort study of all pregnancies complicated by pregestational and gestational DM from 2007-2017 at a Resident Diabetes in Pregnancy clinic at an academic tertiary care center. Patients were excluded if they did not take long acting or intermediate insulin or took an oral agent in addition, had end stage organ damage, multiple gestation, or fetal anomalies. The primary outcome was maternal medication adherence per physician report. The secondary outcome was a composite neonatal outcome including macrosomia, NICU admission, LGA, shoulder dystocia, individual components of the composite, and glycemic control. Demographic characteristics were compared between groups and logistic regression was used to adjust for potential confounders, including advanced maternal age, weight, race, body mass index, and substance abuse.

Results: Of 926 patients seen during the study period, 808 women met study criteria, by diabetes type: type 1 (n=48), type 2 (n=293) and GDM (n=467). Demographic characteristics were similar between those who used long-acting vs. intermediate insulin. Adjusted analysis using logistic regression demonstrated no significant difference in medication adherence between groups. There was also no difference in the neonatal composite outcome, components of the composite, or glycemic control. Results were similar in stratified analysis by diabetes type.

Conclusion: Medication adherence was similar between women using long and intermediate acting insulin and there was no change in neonatal outcomes. Our findings call into question the notion that the twice daily dosing of intermediate insulin in pregnancy provides tighter glycemic control, although medication adherence did not suffer.

Disclosure

S.A. Nazeer: None. C.J. Herrick: None. H.E. Duckham: None. E.B. Carter: None.

Funding

American Diabetes Association/Pathway to Stop Diabetes (1-19-ACE-02 to E.B.C.)
2225-PUB: Direct Actions of the Dual SGLT1/2 Inhibitor Sotagliflozin on Functional Recovery following Global Ischemia in Diabetic and Healthy Mouse Hearts
Authors: DOREY EMILY S. , COTTER BROOKE , MULLINS THOMAS P. , REICHELT MELISSA E. , GALLO LINDA A.
Published: June 2020

Sotagliflozin is a dual sodium-dependent glucose transporter (SGLT) inhibitor with the potential to be cardio-protective in people with and without diabetes. Here, we used an ex vivo Langendorff reperfusion model to investigate if direct infusion with sotagliflozin in hearts from diabetic and healthy mice improves functional recovery from global ischemia. Male C57Bl/6 mice were fed either a control chow (CON) or high fat diet (HFD; 60% of calories from fat) for 46-48 weeks. Fasting blood glucose measurements were performed and body composition measured using nuclear magnetic resonance. Constant pressure Langendorff perfusion experiments were performed on hearts from each diet group. Following equilibration, hearts were infused with either sotagliflozin (90 nM) or dimethyl sulfoxide (VEH) for 10 min prior to a 25 min period of global ischemia. The infusions were recommenced for the first 15 min of 45 min reperfusion. Post ischemic recovery was assessed as % baseline recovery. HFD mice were heavier (54 vs. 34 g) with increased fat mass (42 vs. 19 %) and increased fasting blood glucose (9.6 vs. 8.0 mmol/L). Paradoxically, their functional recovery from global ischemia was, in general, better when compared with hearts from CON mice and there was no effect of sotagliflozin infusion. In CON mice, early post-ischemic coronary flow was greater with sotagliflozin infusion when compared with VEH. Functional recovery in dP/dtmax and dP/dtmin was also greater with sotagliflozin infusion when compared with VEH.

In conclusion, HFD exerted a protective effect on heart function, despite inducing obesity and hyperglycemia. Ongoing studies are examining the effects of sotagliflozin in an alternative model of T2DM. In lean mice, sotagliflozin increased functional recovery from global ischemia, highlighting the potential benefits of an antidiabetic agent in acute coronary care.

Disclosure

E.S. Dorey: None. B. Cotter: None. T.P. Mullins: None. M.E. Reichelt: None. L.A. Gallo: None.

Funding

Diabetes Australia
2228-PUB: Utilizing a Diverse Stakeholder Group to Build a Patient-Centered Type 1 Diabetes Research Agenda
Authors: JEWELL VANESSA D. , ABBOTT AMY A.
Published: June 2020

Engaging patient’s in the research process is essential as it creates self-awareness, increases a collective understanding of needs and views, and allows for active participation in healthcare decisions. This stakeholder driven project aimed to develop a patient-centered research model to identify stakeholders of type 1 diabetes (T1D) for addressing the needs of patients and families in rural communities. From project inception, investigators used an innovative approach of incorporating stakeholders in the entire research process. A four-step engagement plan using the Methods in Community-Based Participatory Research for Health guided the project. In the first strategy, assessment of both personal and institutional capacities for active collaboration was completed. Specifically, the project lead (researcher, healthcare provider, and mother of a child with type 1 diabetes), caregiver, and advocacy organization executive met to discuss strengths and weaknesses and how to effectively collaborate. Next, identification of potential partners ensued with new and established networks, associations, and community leaders. We completed this step through years of networking and becoming involved within the community. Reframing the ultimate health issue for research was the third strategy. This was a dynamic, interactive process guided by a diverse group of stakeholder perspectives across two states in the development of a patient-centered research plan. Finally, developing and nurturing the sustained partnerships established between researchers and community partners was accomplished in strategy four. Specifically, the team developed a group process contract that allowed for fluctuation to meet individual and group needs. These relationships were built through sustained T1D community presence and networking. This shared decision-making process resulted in trust, accountability, and stakeholder empowerment while ensuring that individual’s voices were heard.

Disclosure

V.D. Jewell: None. A.A. Abbott: None.

Funding

Patient-Centered Outcomes Research Institute (10443-CU)
2235-PUB: Improving Rates of Monitoring for Type 2 Diabetes Complications
Authors: SINGH PUJA , WOLFGRAM PETER
Published: June 2020

Introduction: To compare our diabetes clinic’s baseline adherence to American Diabetes Association (ADA) and International Society of Pediatric and Adolescent (ISPAD) guidelines for obtaining urine creatinine/albumin ratio (ACR), liver enzymes (LFTs) and lipids at diagnosis with type 2 diabetes (T2D) to the rates of screening adherence following institution of educational flowcharts, revised note templates, and a best practice advisory (BPA).

Methods: Data was extracted from Epic including all patients seen initially in our diabetes outpatient clinic with recently diagnosed T2D from 7/2014-8/2019. Data included dates of initial patient visits to our clinic and dates of ACR, LFTs, and lipid order placement or previous lab results. Vizio flowcharts and a revised clinic note template reflecting ADA/ISPAD guidelines were created and presented to the clinic’s 8 attendings and 5 APNPs in 7/2016. In 10/2017, a BPA alert containing the recommended labs began firing if T2D was on the patient’s problem list. A provider was considered adherent to the guidelines if labs were available from the previous 6 months or ordered within 120 days of diagnosis. Pre and post-screening rates were compared using the t-test.

Results: Analysis included 40 patients diagnosed with T2D before any interventions and 39 after the 10/2017 BPA began. The improvement in urine ACR ordering rate was significant as it went from a baseline of 32.5% up to 82.1% (p<0.001), while improvements in the LFT ordering rate (60.0% up to 74.4%; p=0.2) and the lipid order rate (70.0% up to 79.0%; p=0.4) were not significant.

Conclusion: With the implementation of the T2D flowchart, templated note, and BPA, adherence to ADA/ISPAD of ordering baseline screening labs rose to the 70-80% range. Given our aim to detect and treat complication as early as possible, and in order to increase reliability of our care, we are going to begin posting quarterly run charts so both the ordering provider and the diabetes nurses are consistently updated on our adherence rates.

Disclosure

P. Singh: None. P. Wolfgram: None.
2240-PUB: Predictability of HbA1c Control Based on Pharmacotherapy Regimen or the Number of Diabetes-Related Touches in a Rural Population
Authors: VASCIMINI ANGELINA , SULLIVAN TARA , CURTIS STACEY D. , DEREMER CHRISTINA E.
Published: June 2020

Given the explosion of pharmacologic agents indicated for patients diagnosed with type 2 diabetes, coupled with the decline in physicians practicing in rural settings, pharmacists are positioned to serve as extenders. In this resource-limited environment, assumptions are often applied to predict outcomes based on patient’s frequency of appointments or pharmacotherapy regimen. The goal of this study is to determine if select medications or the number of clinical touches support assumptions predicting a patient’s HbA1c control.

A single center retrospective chart review was conducted for patients with an ICD-10 code related to type 2 diabetes. Data extracted from the electronic medical record included demographics, HbA1c with trend, dates of diagnosis, pharmacotherapy regimen, and number of clinical diabetes related touches. Touches were defined as documentation of diabetes related management through office appointment or remote communication. This project was approved by the institutional quality improvement performance registry process. Fisher exact, Wilcoxon test, and descriptive statistics were used for analysis.

Sample data for 300 patients revealed baseline demographic information for study population’s average age of 64 years and 43% were female. The number of clinical touches on predicting HbA1c control was not found to be statistically significant (P = 0.1582). However, there was a significant association for select medications when evaluated in combination with frequency of clinical touches to predict control. Insulin was the only agent that when evaluated solely resulted in a significantly lower HbA1c (p=0.03).

Sample population data shows that increased touches did not predict improved HbA1c values, however it could be considered that less controlled HbA1c were being followed more frequently and thus altering outcomes. Additional evaluation is needed to more closely investigate the combination of agents.

Disclosure

A. Vascimini: None. T. Sullivan: None. S.D. Curtis: None. C.E. DeRemer: Advisory Panel; Self; Bristol-Myers Squibb. Speaker’s Bureau; Self; Bristol-Myers Squibb, Pfizer Foundation. Stock/Shareholder; Self; Portola Pharmaceuticals, Inc.
2252-PUB: Fetal Cardiac Evaluation by Ultrasound to Assess Pregnancies Complicated by Maternal Diabetes
Authors: LEE-TANNOCK ALISON , HAY KAREN , KUMAR SAILESH
Published: June 2020

Objectives: To evaluate fetal and neonatal cardiac morphology by echocardiography in pregnancies complicated by diabetes compared to normal healthy controls using ultrasound.

Methods: Fetal echocardiography was performed in 121 women with uncomplicated singleton pregnancies and 50 women with diabetes in pregnancy. The women underwent ultrasound scans every four weeks from enrolment (between 18-28 weeks gestational age) until delivery. Neonatal echocardiography was performed on the offspring. M-mode assessment was used to assess interventricular septal (IVS) thickness, left ventricle diastolic dimensions (LVED), fractional shortening (FS), tricuspid annular plane systolic excursion (TAPSE) and mitral annular plane systolic excursion (MAPSE).

Results: There were differences in IVS thickness, LVED, FS and TAPSE values between women with uncomplicated pregnancies and women with diabetes in pregnancy. These differences persisted into the neonatal period.

Conclusion: M-mode is a simple ultrasound technique to assess fetal cardiac function. Maternal diabetes was associated with increased IVS thickness, and reduction in TAPSE and FS measures. Prognostic implications of these findings may be useful in assessing pregnancies complicated by maternal diabetes in addition to current ultrasound assessment of fetal size and Doppler studies of fetal vessels.

Disclosure

A. Lee-Tannock: None. K. Hay: None. S. Kumar: None.

Funding

Mater Health Services; Australasian Society for Ultrasound in Medicine
2256-PUB: Clonal Hematopoiesis of Indeterminate Potential (CHIP), Glycemic Traits, and Type 2 Diabetes (T2D) Risk
Authors: TOBIAS DEIRDRE K. , RAGHAVAN SRIDHARAN , BICK ALEXANDER , RAFFIELD LAURA , WU PEITAO , DICORPO DANIEL A. , DUPUIS JOSEE , LANGE LESLIE , ROTTER JEROME I. , KOOPERBERG CHARLES L. , MEIGS JAMES B. , MANSON JOANN E.
Published: June 2020

Background and Objective: CHIP is the aging-related accumulation of genetic mutations in the hematopoiesis-associated genes of stem cells. CHIP was recently associated with total mortality and atherosclerotic cardiovascular disease. We tested the hypothesis that CHIP is associated with adverse glycemic traits and incident T2D risk.

Methods: This study includes a subset of n=9,704 individuals in the NHLBI Trans-omics for Precision Medicine (TOPMed) project from the Framingham Heart Study (FHS), Jackson Heart Study (JHS), Multi-Ethnic Study of Atherosclerosis (MESA) and Women’s Health Initiative (WHI). We analyzed individuals without a history of T2D, cancer, or cardiovascular disease, and collected blood for fasting glycemic traits and CHIP carrier status. CHIP was recently determined from blood DNA-derived whole genome sequencing using the CATK MuTECT2 somatic variant caller in pre-specified leukemogenic driver mutations in candidate clonal expansion genes. We are conducting multivariable linear models examining both cross-sectional associations of CHIP with glycemic traits and survival models for baseline CHIP with incident T2D, adjusting for age, sex, body mass index, smoking, cohort, and family history of diabetes.

Results: Mean (SD) age was FHS = 50 (14), JHS = 55 (13), MESA = 60 (10), and WHI = 66 (7) years. We identified n=460 (4.7%) as CHIP carriers. The most common variant was located in DNMT3A gene (60% of carriers). Among CHIP carriers, the prevalence of having CHIP present in 1 of the 4 genes previously implicated in atherosclerotic heart disease (DNMT3A, TET2, ASXL1, JAK2; Jaiswal, et al, NEJM 2017) was lowest in FHS (75%) and highest in JHS (96%). Results for CHIP in relation to glycemic traits and incident T2D (∼1,500 cases) will be presented in detail. We will additionally present the associations for gene-specific CHIP mutations.

Conclusions: These cutting-edge TOPMed data will elucidate the role of CHIP, a novel aging biomarker, in relation to T2D risk.

Disclosure

D.K. Tobias: None. S. Raghavan: None. A. Bick: None. L. Raffield: None. P. Wu: None. D.A. DiCorpo: None. J. Dupuis: Consultant; Self; Merck Sharp & Dohme Corp. L. Lange: None. J.I. Rotter: None. C.L. Kooperberg: None. J.B. Meigs: Consultant; Self; Quest Diagnostics. J.E. Manson: None.

Funding

National Institute of Diabetes and Digestive and Kidney Diseases (U01DK078616 to J.B.M., J.D., D.A.D., P.W.), (U01DK105554 to J.D., D.A.D.)