bluuebunny/crossref_metadata_2025 huggingface dataset:
1568-P: One Methylation Risk Score Model, Two Racial Populations
Authors: MINNIEFIELD BRE A. , IRVIN MARGUERITE , HIDALGO BERTHA , TIWARI HEMANT K. , PATKI AMIT , CHAUDHARY NINAD
Published: June 2020

Metabolic Syndrome (MetS) is a cluster of conditions that can increase the risk of chronic diseases like, type 2 diabetes. Multiple reports show alterations in DNA methylation patterns are associated with the condition. Using cross-sectional data from Hypertension Genetic Epidemiology Network (HyperGEN) (N=614 African Americans (AA)) and Genetics of Lipid Lowering Drugs and Diet Network (GOLDN) (N=995 European Americans (EA)), we created a risk score model for MetS. First, we selected specific cytosine-guanine dinucleotides (CpG) sites measured on Illumina Methyl450 arrays previously found to be significantly associated to MetS (CPT1A cg00574958, PHOSPHO1 cg02650017, ABCG1 cg06500161, SREBF1 cg11024682, SOCS3 cg18181703, TXNIP cg19693031). Using the HyperGEN parameter estimates for the single CpGs as weights, we calculated a methylation risk score in HyperGEN which was validated in GOLDN. We then conducted an association analysis using a generalized linear mixed model to test the association between the methylation risk score (MetS-score) and MetS, adjusting for age, sex, study site, 4 ancestry principal components, and family structure. In HyperGEN we performed a 100,000-permutation test to determine statistical significance of the score with MetS. Results showed the risk score was significantly associated with MetS in both populations (HyperGEN p=6.36x-10; GOLDN p=2x-16), with a 1 standard deviation increase in the score being associated with 2.25 (95% CI, 1.79 - 2.86) higher odds of having MetS in HyperGEN and a 2.45 (CI 95%, 2.02 - 3.00) higher odds of having MetS in GOLDN. Lastly, we compared the model fitness of the GOLDN MetS model with and without the MetS-score. AIC criterion suggested the model including the score was a better fit (no score = 1173.99; with score = 1112.61).

In conclusion, we created a methylation risk score strongly associated with MetS which should be further investigated for its ability to predict MetS risk in both AAs and EAs.

Disclosure

B.A. Minniefield: None. M. Irvin: None. B. Hidalgo: None. H.K. Tiwari: None. A. Patki: None. N. Chaudhary: None.

Funding

National Heart, Lung, and Blood Institute (3R01HL123782-02S1)
1587-P: The Impact of Cannabis Use on Glucose Tolerance
Authors: ALSHAARAWY OMAYMA
Published: June 2020

Cannabis use prevalence has increased in the United States (U.S.) and worldwide. Cannabis exerts its effects on human function primarily through activating the cannabinoid system. In recent years, the cannabinoid system has emerged as an important regulator of energy balance. Pharmacological deactivation or genetic deletion of the cannabinoid-1 receptor in animals improved various metabolic outcomes. Human epidemiological studies, however, suggests a paradox. Increased caloric intake and decreased physical activity were observed in cannabis users. Surprisingly, cannabis use was associated with attenuated weight gain and lower levels of fasting insulin. The aim of this study is to examine the association of cannabis use and glucose clearance as indicated by glucose tolerance test among adults participating in the National Health and Nutrition Examination Survey (NHANES). NHANES is designed to recruit, interview and examine a nationally representative sample of the U.S. non-institutionalized population. For this study, we used publicly available NHANES data generated through surveys conducted in 2007-2016. After a 9-hour fast, a subsample of participants were asked to drink a calibrated dose (75 grams of glucose) of TrutolTM and had a second venipuncture 2 hours after drinking. Cannabis use questions were self-administered using the Audio Computer-Assisted Self-Interview in NHANES mobile examination center. Results from generalized regression modelling indicated an inverse association between cannabis use in the 30 days prior to NHANES and 2-hr blood glucose levels (β= -5.9; 95% CI = -9.9, -1.8). Adjusting for tobacco smoking, alcohol drinking or body mass index minimally affected the estimates. The relationship between 2-hr glucose levels and cannabis use frequency was stronger for occasional users compared to heavy users.

In conclusion, cannabis use is associated with improved glucose clearance. Additional studies are needed to confirm the results of this study and to investigate the underlying mechanisms.

Disclosure

O. Alshaarawy: None.

Funding

National Center for Complementary and Integrative Health (AT009156)
1601-P: Cardiovascular Risk Profiles among Women by Gestational Diabetes Status, 2007-2016
Authors: DORANS KIRSTEN S. , BAZZANO LYDIA A. , HE JIANG
Published: June 2020

Background: Women with prior gestational diabetes (GDM) are at elevated risk of type 2 diabetes and cardiovascular disease. We sought to compare cardiometabolic risk factors among parous U.S. women ages 20 to 44 by history of GDM.

Methods: We used data from the National Health and Nutrition Examination Survey (NHANES) from 2007-2016. We included 3562 parous women. We compared anthropometric measures, glycemic markers, blood pressure and lipids by self-reported history of GDM, both with and without adjusting for demographic and behavioral factors and waist circumference. Analyses took NHANES multi-stage survey design into account.

Results: Women without history of GDM were younger and, in crude analyses, had more favorable cardiometabolic risk factor profile than women with history of GDM (Table). Similar patterns were observed after adjusting for age, race/ethnicity, education and smoking. After also adjusting for waist circumference, only hemoglobin A1c, fasting glucose, high-density lipoprotein cholesterol and low-density lipoprotein cholesterol were significantly higher among women with history of GDM compared to women without GDM.

Conclusions: Among parous U.S. women ages 20 to 44, those with a history of GDM had less favorable cardiometabolic risk factor profiles than did women without GDM, though results were attenuated after adjusting for markers of demographics, behavior, and central obesity.

Disclosure

K.S. Dorans: None. L.A. Bazzano: None. J. He: None.

Funding

Tulane Center of Biomedical Research (P20GM109036)
1624-P: Family History of Type 1 Diabetes (T1D) and Comorbidity Status in Individuals with T1D: A Computational Deep Exploratory Data Mining Approach
Authors: TALLON ERIN M. , CLEMENTS MARK A. , LIU DANLU , BOLES KATRINA , STUCK RACHEL A. , SHYU CHI REN
Published: June 2020

T1D is a progressive, autoimmune-mediated condition associated with numerous comorbidities and significant premature mortality. The offspring of fathers diagnosed with T1D exhibit a 2-3 times higher risk of developing T1D than the offspring of mothers diagnosed with T1D. Individuals with T1D who have an affected father present with a more aggressive disease phenotype at diagnosis. Yet the impact of paternal T1D (P-T1D) vs. maternal T1D (M-T1D) on health outcomes in individuals with T1D is unknown.

We previously developed and validated a subgroup discovery algorithm that identifies highly contrasted patterns occurring with a significant difference in prevalence between two subgroups (e.g., P-T1D vs. M-T1D). Here we used this computational deep exploratory data mining method to analyze publicly-accessible data from the T1D Exchange Clinic Registry (2010-12; 2015-17). We used an Apache Spark high performance computing environment to apply our algorithm to family history, gender, and medical conditions data in the Registry.

We identified multiple phenotypic contrasts between individuals with P-T1D (2010-12 data: n = 1011; 2015-17 data: n = 652) vs. M-T1D (2010-12 data: n = 528; 2015-17 data: n = 342). Using Fisher’s exact tests to determine statistical significance, P-T1D associated with hypothyroidism and male gender in 7.2% of cases (47/652), compared to 3.2% of cases (11/342) of M-T1D (p = 0.01). M-T1D was associated with hypertension (HTN) in 36.0% of cases (123/342), compared to 22.3% of cases (146/652) in P-T1D (p < 0.001). M-T1D was associated with diabetes-related neuropathy and HTN in 10.8% of cases (37/342), compared to 4.9% of cases (32/652) in P-T1D (p = 0.001).

Complex patterns generated with this algorithm may yield contrast patterns that provide insights about personalized interventions to improve health outcomes. Whether contrast mining can be used to predict health trajectories of individuals with T1D remains to be determined.

Disclosure

E.M. Tallon: None. M.A. Clements: Consultant; Self; Glooko, Inc. Other Relationship; Self; Glooko, Inc. D. Liu: None. K. Boles: None. R.A. Stuck: None. C. Shyu: None.

Funding

National Institutes of Health (5T32LM012410)
1627-P: Real-World Comparison of Acute Complications in Adult Patients with Type 1 Diabetes in U.S. and European Registries
Authors: ZHOU FANG L. , HERMANN JULIA , PETTUS JEREMY , KERNER WOLFGANG , MILLER KELLEE , STECHEMESSER LARS , EDELMAN STEVEN , IBALD-MULLI ANGELA M. , HOLL REINHARD W.
Published: June 2020

The study compares acute complications among adults with type 1 diabetes (T1D) by glycemic control in a U.S. Electronic Health Record database (T1PCO) with the German/Austrian Prospective Diabetes Follow-up (DPV). T1PCO included 31,430 and DPV 5,190 individuals in the analysis (age ≥18 years, T1D duration ≥2 years). During baseline (12 months prior to index date), ≥1 HbA1c and ≥1 insulin prescription was required. Baseline demographics, severe hypoglycemia (SH) and diabetic ketoacidosis (DKA) were analyzed for overall populations, by age group, and glycemic control (HbA1c categories). In both databases, incidence of DKA was highest in those with HbA1c ≥9.0%. Additionally, the incidence of DKA (per year) increased with deteriorating control (HbA1c<8%: 1.3% vs. 1.7%, HbA1c≥10%: 19.9% vs. 22.9%; DPV and T1PCO respectively). In contrast, incidence of SH patterns differed between the databases. While in the T1PCO cohort SH increased with poorer glycemic control, the trend was reversed in DPV cohort (Figure). Real-world comparisons demonstrate the challenges in diabetes care across many countries around the world. Different patterns of incidence of SH by HbA1c may be related to differences in the capture of SH in databases, it may also reflect important differences in diabetes management including access to different types of insulin, CGM, pumps, education and type 1 savvy health care providers.

Disclosure

F.L. Zhou: Employee; Self; Sanofi. Stock/Shareholder; Self; Sanofi. J. Hermann: None. J. Pettus: Consultant; Self; Diasome Pharmaceuticals, Inc., Lexicon Pharmaceuticals, Inc., MannKind Corporation, Novo Nordisk Inc., Sanofi. W. Kerner: None. K. Miller: None. L. Stechemesser: None. S. Edelman: Advisory Panel; Self; Brightsight, InPen, Lexicon Pharmaceuticals, Inc., Lilly Diabetes, Novo Nordisk A/S. Board Member; Self; Senseonics. Speaker’s Bureau; Self; AstraZeneca, MannKind Corporation, Merck & Co., Inc., Sanofi. A.M. Ibald-Mulli: Employee; Self; Sanofi-Aventis. R.W. Holl: None.

Funding

Sanofi
1633-P: Women with Type 1 Diabetes (T1D) Experience a Shorter Reproductive Period Compared with Nondiabetic Women: The Pittsburgh Epidemiology of Diabetes Complications (EDC) Study and the Study of Women’s Health Across the Nation (SWAN)
Authors: YI YAN , EL KHOUDARY SAMAR R. , MILLER RACHEL G. , RUBINSTEIN DEBRA , MATTHEWS KAREN A. , ORCHARD TREVOR J. , COSTACOU TINA
Published: June 2020

Evidence suggests that insulin deficiency and hyperglycemia may disrupt the normal function of the female reproductive system, leading to delayed menarche and premature ovarian aging. We compared the length of the reproductive period of women with T1D and women without diabetes. We assessed age at menarche and natural menopause in women with childhood-onset T1D (diagnosed in 1950-80) who participated in the Pittsburgh EDC study’s 2010-16 follow-up (where reproductive history was assessed) and nondiabetic participants from the Pittsburgh site of the SWAN study. Women who had not yet reached natural menopause, had a hysterectomy/oophorectomy before menopause or received sex hormone therapy during the menopausal transition were excluded from analyses. In EDC, covariate data were selected from the follow-up visit where chronological age was closest to the baseline age in SWAN. Reproductive history was self-reported. The Women’s Ischemia Syndrome Evaluation historical and hormonal algorithms were also used to assess menopause status. Women with T1D (n=105) were younger (42.8 vs. 46.0 yrs, p=.0003), more likely to be white (96.2% vs. 68.3%, p<.0001), never smokers (67.3% vs. 48.1%, p=.002), with lower BMI (25.2 vs. 26.5 kg/m2, p=.002) and higher HDL-C (61.0 vs. 53.0 mg/dl, p<.0001) compared with women without diabetes (n=341). Women with T1D were also older at menarche (13.3 vs. 12.6 yrs, p<.0001) but younger at natural menopause (50.1 vs. 51.9 yrs, p<.0001). Adjusting for age, race, smoking status, BMI, HDL, and number of pregnancies, T1D was associated with 3.3 (95% CI: 2.5-4.0) fewer reproductive years. The length of the reproductive period of women with T1D is shorter, having delayed menarche and earlier natural menopause, compared with women without diabetes. Factors that could be related to a shorter reproductive period in T1D should be investigated.

Disclosure

Y. Yi: None. S.R. El Khoudary: None. R.G. Miller: None. D. Rubinstein: None. K.A. Matthews: None. T.J. Orchard: None. T. Costacou: None.

Funding

National Institutes of Health (DK34818); Rossi Memorial Fund; National Institute on Aging; National Institute of Nursing Research; Office of Research on Women’s Health
1663-P: Gene Editing of the Human T-Cell Receptor Locus Enables Optimized In Vitro Testing of Autoreactive T-Cell Function in Type 1 Diabetes
Authors: PETERS LEEANA D. , POSGAI AMANDA L. , BRUSKO TODD M.
Published: June 2020

Gene editing of the human T cell receptor locus enables optimized in vitro testing of autoreactive T cell function in type 1 diabetes.

The generation of antigen specific T cells and subsequent implementation as “living drugs” has garnered great interest in the immunotherapy space. One of the main caveats to this approach is the potential for heterologous chain pairing between the α- and β-chains of the endogenous and de-novo T cell receptors (TCRs). The consequences of chain mispairing can range from reduced antigen specific functionality to off-target autoimmune reactivity. The latter is an important consideration when designing therapies for autoimmune and inflammatory diseases, specifically with regards to the introduction of regulatory T cells (Treg) adoptive cell therapy. To date, infusions of polyclonal Tregs have demonstrated safety but have failed to reverse established type 1 diabetes (T1D). This demonstrates the need for the generation of safe and effective antigen-specific Tregs capable of recognizing β-cells in T1D. We have previously shown the ability to generate GAD-specific Tregs which maintain lineage stability and suppressive phenotype. To extend this work, we deleted the endogenous TCR α-chain gene (TRAC) via CRISPR/cas9 in CD8+T cells. We achieved >90% editing efficiency through ribonucleoprotein (RNP) complex mediated single guide RNA delivery. Moreover, we found TRAC-KO CD8+ T cells displayed increased peptide MHC-dextramer staining, CD69 and CD25 upregulation after stimulation with cognate antigen, and increased antigen-specific killing of β-cell lines when compared to mock guide treated T cells. We believe this workflow can be applied to generate antigen specific Tregs, which we expect to provide increased suppressive capacity. Hence, we present a TCR-gene editing protocol in primary human T cells that results in efficient deletion of the endogenous TCR α-chain and robust enhancement of antigen-specific T cell function.

Disclosure

L.D. Peters: None. A.L. Posgai: None. T.M. Brusko: Board Member; Self; OneVax, LLC. Consultant; Self; Caladrius Biosciences, Inc., Sanofi-Aventis. Speaker’s Bureau; Self; CSL Behring.

Funding

National Institutes of Health (P01AI42288, R01DK106191, T325T32DK108736-03HIRN, UC4 P0063595)
1672-P: Characterization of Immune Checkpoint Inhibitor–Mediated Hyperglycemia: Beyond Insulin-Dependent Diabetes
Authors: LEITER AMANDA , CARROLL EMILY , BROOKS DANIELLE C. , SHIMOL JENNIFER BEN , EISENBERG ELLIOT , GALSKY MATTHEW , GALLAGHER EMILY
Published: June 2020

Immune checkpoint inhibitors (ICIs) treat cancer by augmenting patients’ (pts) immune responses against cancer. The immune activation from ICIs is associated with immune-related adverse events. Inflammation has a known association with hyperglycemia (HG) and insulin resistance. Little is known about the prevalence of post-ICI HG (pIHG) beyond autoimmune diabetes (DM). We hypothesized that: (1)Pts with pIHG are more likely to have steroid exposure, pre-existing DM, and obesity (body mass index ≥30kg/m2); (2)A subset of pts have pIHG that cannot be explained by autoimmune DM or steroid use. We analyzed a cohort of 411 cancer pts on ICIs between 2011 and 2017 at our institution. We included pts who had ≥3 post-ICI glucose measurements. We defined pIHG as a random glucose >200mg/dL from ICI initiation up to 6 months after cessation. We recorded DM diagnosis and steroid exposure as documented in the medical record. We compared pts with and without pIHG using the Fisher’s exact test. 385 pts had post-ICI glucose data. 105 (27.3%) had pIHG. Compared to those without pIHG, pIHG pts were more likely to be male (71% vs. 59%, p=0.04), to have pre-existing DM (54% vs. 10%, p<0.01) and to be obese (31% vs. 18%, p<0.01). Of the 105 pts with pIHG, 59(56%) had pre-ICI HG, 9(8.6%) had no pre-ICI glucose data, and 37(35.2%) had new onset pIHG. Of these 37 pts, 25(67.6%) had steroid-related HG and none had antibody-confirmed autoimmune DM. Of the remaining 12 pts, 3 had worsening of pre-existing DM. No obvious precipitant of HG was detected in the remaining 9 pts apart from ICI therapy. Timing of HG after ICI initiation ranged from 4 to 107 weeks (median 12.3 weeks). HG in 5 of these pts resolved without intervention.

In conclusion, pIHG occurred in over a quarter of pts. Risk factors for pIHG included pre-existing DM and obesity. The majority of new pIHG was related to steroid use. However, a subset of pts had new HG that may be related to ICI therapy, suggesting that ICIs may have a role in immune-related insulin resistance.

Disclosure

A. Leiter: None. E. Carroll: None. D.C. Brooks: None. J. Ben Shimol: None. E. Eisenberg: None. M. Galsky: Advisory Panel; Self; AstraZeneca, Bristol-Myers Squibb, Genentech, Inc., Merck & Co., Inc., Pfizer Inc. E. Gallagher: Advisory Panel; Self; Novartis Pharmaceuticals Corporation.

Funding

National Cancer Institute (P30CA196521, K08CA190770)
1674-P: Stearate Potentiates MIP-1a Production with TNF-a via TLR4/TBK1/IKKe /IRF3 Signal Axis: Significance for Insulin Resistance
Authors: AHMAD RASHEED , KOCHUMON SHIHAB P. , AREFANIAN HOSSEIN , ALMULLA FAHD
Published: June 2020

Increased levels of plasma free fatty acids (FAs) are an important contributor to chronic inflammation, insulin resistance and other obesity-related complications. However, the molecular mechanisms by which individual FAs contribute to obesity-associated inflammation and disorders remain incompletely understood. Here we report that dietary saturated FAs specifically stearate led to a profound production of macrophage inflammatory protein (MIP)-1α (CCL3) in the presence of TNF-α in monocytes, macrophages and adipose tissues. Using pharmacologic and genetic approaches, we identified the involvement of TLR4/TBK1/IKKε/IRF3 signal axis in this production of MIP-1α in response to stearate and TNF-α. Consistent with this, cultured adipose tissues from TLR4 knockout (KO) mice revealed that stearate and TNF-α treatment did not induce MIP-1α compared with TLR2 KO or wild type (C57BL/6) mice. Moreover, C57BL/6 mice fed a high fat diet (HFD) for 16 weeks showed elevated levels of plasma fatty acids, TNF-α and MIP-1α along with insulin resistance. MIP-1α was significantly correlated with TNF-α. Our human data shows that expression of TLR4/p-IRF3 was elevated in the PBMCs of obese individuals as compared to lean. Furthermore, elevated MIP-1α expression levels in obese fat tissues were significantly correlated with TNF-α and macrophage markers.

In conclusion, these findings provide a mechanistic link between stearate and TNF-α for the overproduction of obesity associated MIP-1α, which could be used as a new target for the treatment of obesity-related chronic inflammation/insulin resistance.

Disclosure

S.P. Kochumon: None. F. Almulla: None.

Funding

Kuwait Foundation for the Advancement of Science (RA-AH-2016-007)
1678-P: Insulin Peptide Reactive T-Cells May Reflect Disease Activity in Ketosis-Prone Type 2 Diabetes
Authors: SATOMURA ATSUSHI , SUZUKI SEIYA , HAISA AKIFUMI , OIKAWA YOICHI , SHIMADA AKIRA
Published: June 2020

Ketosis-prone type 2 diabetes with autoantibody negative and beta cell function positive group (KPD) is recognized as atypical diabetes characterized by the onset of disease at a relatively young age, obesity, sudden onset of ketosis and a transient decrease in insulin secretion. Previously we have reported that insulin peptide reactive T cells are detected in KPD patients (2017, ADA meeting). In this study, we examined the correlation between insulin peptide reactive T cells and clinical background such as C-peptide, HbA1c, and HLA types especially focusing on class I region to assess the significance of insulin peptide reactive T cells detected in KPD subjects. Peripheral blood was obtained from enrolled subjects with informed consent, and stimulated with insulin peptides (insulin B9-23, B10-24, B11-25, B12-26) for 24 hours, then interferon (IFN)-gamma spots were counted. Maximum spots more than 2.5 were considered as positive based on the data from control subjects. Seven out of 21 (33.3%) KPD participants (21 males, 0 females, mean age 40.3 years) showed positive result, and this frequency was similar to that in acute-onset type 1 diabetes (17 out of 50; 34.0%) (22 males, 28 females, mean age 44.5 years); these results were consistent with our previous observation. Only 1 out of 35 (2.9%) showed positive result in controls (26 males, 9 females, mean age 61.8 years). In KPD participants, serum C-peptide level was significantly lower in the ELISpot positive group than in the negative group (1.40±0.55 ng/mL vs. 2.35±0.76 ng/mL; p <0.05). Moreover, there was a significant positive correlation between the spot number and HbA1c at onset in the positive group (rs= 0.6232, (p <0.05 by Spearman correlation test). Furthermore, HLA A*02:01 or A*24:02 positive rate was significantly higher in the ELISpot positive group (n=7) than the negative group (n=14) (86% vs. 36%; p <0.05). These findings suggest that insulin peptide reactive IFN-gamma-producing T cells may reflect disease activity of KPD.

Disclosure

A. Satomura: None. S. Suzuki: None. A. Haisa: None. Y. Oikawa: None. A. Shimada: Advisory Panel; Self; Astellas Pharma Inc. Speaker’s Bureau; Self; Eli Lilly Japan K.K., Novo Nordisk Inc., Sanofi-Aventis.
1699-P: Mechanisms and Roles of Foxo1-Akt Negative Feedback Pathway in Adipocytes
Authors: OHNO TOMOHIRO , ONO HIRAKU , YOKOTE KOUTARO
Published: June 2020

Objective: FoxO1 is a substrate of serine/threonine-kinase Akt. Although FoxO1 is phosphorylated by Akt and inactivated by insulin stimulation, constitutive activation of FoxO1 is reported to enhance Akt phosphorylation. Therefore, constitutive activation of FoxO1 may mimic insulin’s functions via Akt phosphorylation.

Methods: Constitutively-active FoxO1 (CA-FoxO1) in 3T3-L1 adipocytes is overexpressed by adenovirus and Changes in insulin signaling and downstream actions was studied.

Results: CA-FoxO1 induced Akt phosphorylation in differentiated 3T3-L1 adipocytes. Regarding downstream actions, CA-FoxO1 significantly enhanced glucose uptake (148.6% compared to controls, P =0.030), GLUT4 translocation (297.6%, P <0.01), lipogenesis (175.9%, P =0.030) and tended to enhance glycogen synthesis (128.7%, P =0.154), while in contrast, under insulin-stimulated condition, significantly suppressed protein synthesis (56.5%, P <0.01) and DNA synthesis (44.4%, P <0.01). Regarding insulin signaling, CA-FoxO1 enhanced phosphorylations of GSK-3, PRAS40, and p70S6K. The enhancement of PI 3-kinase activity by 3nM insulin is larger than that by CA-FoxO1, while Akt phosphorylation by 3nM insulin is smaller than that by CA-FoxO1, suggesting the mechanism of FoxO1-Akt feedback is downstream of PI 3-kinase. In contrast to previous papers, protein amounts of IR-β, p110α subunit of PI 3-kinase, or Rictor were not significantly changed by CA-FoxO1.

Conclusion: Constitutive activation of FoxO1 induced Akt phosphorylation even in differentiated 3T3-L1 adipocytes, but FoxO1 activation mimicked only metabolic functions of glucose but not proliferative or oncogenic functions of insulin. This selective mimicking of insulin by FoxO1 activation in adipocytes may improve glucose metabolism with a low risk of tumorigenesis. The mechanisms of FoxO1-Akt feedback is suggested not to be the increase in IR-β or p110α subunit of PI 3-kinase as previously reported, but to be mainly downstream of PI 3-kinase.

Disclosure

T. Ohno: None. H. Ono: None. K. Yokote: Research Support; Self; Astellas Pharma Inc., Daiichi Sankyo, Eli Lilly Japan K.K., Kowa Company, Ltd., Kyowa Hakko Kirin Co., Ltd., Merck Sharp & Dohme Corp., Mitsubishi Tanabe Pharma Corporation, Mochida Pharmaceutical Co., Ltd., Nippon Boehringer Ingelheim Co. Ltd., Novartis Pharma K.K., Novo Nordisk Inc., Ono Pharmaceutical Co., Ltd., Pfizer Japan Inc., Sanofi K.K., Shionogi & Co., Ltd., Sumitomo Dainippon Pharma Co., Ltd., Taisho Pharmaceutical Co., Ltd., Takeda Pharmaceutical Company Limited, Teijin Pharma Limited. Speaker’s Bureau; Self; Abbott, Astellas Amgen, AstraZeneca K.K., Bayer Inc., FUJIFILM Pharmaceuticals U.S.A., Inc., Kaken Pharmaceutical Co., Ltd., Sanwa Kagaku Kenkyusho.
170-LB: Predictors of Diabetes First Diagnosed in Pregnancy: A Machine-Learning Model
Authors: DE CAMPOS MARIA A. , OPPERMANN MARIA LUCIA R. , GENRO VANESSA K. , LEITAO CRISTIANE B. , HIRAKATA VÂNIA N. , REICHELT ANGELA J.
Published: June 2020

Introduction: Pregestacional (PG) DM first diagnosed in pregnancy (diabetes in pregnancy, DIP) carries poor outcomes, which may be prevented by DM diagnosis before pregnancy. Aim: To identify risk factors (RF) for DIP that may help to diagnose DM before pregnancy.

Methods: Retrospective cohort, type 2 DM women; specialized prenatal care facility; public hospitals; Brazil. Data - age, ethnicity, schooling, DM family history (FH), previous obstetric history (number of pregnancies, GDM, preeclampsia (PE), macrosomia) - and PG BMI were collected from May 2005 to July 2019. Univariable analyses (student T and chi-square tests) with SPSS vs.18; two machine learning (ML) models (model 1: all variables; model 2: age, GDM, FH, number of pregnancies and PG BMI) with software Orange Data Mining vs. 3; random sampling statistics.

Results: Of the 466 women, 157 (33.7%, 95% CI 29-38%) had DIP. Baseline characteristics/main outcomes were similar between DIP and PG DM women. For model 1, AUC was 0.526 and precision, 0.566. For model 2 (Figure), AUC was 0.525 and precision, 0.556. Age was the most important RF, followed by previous macrosomia in women ≤ 28 years and by PG BMI in those > 28 years.

Conclusion: In type 2 women, RF identified by ML with the algorithms used were poor predictors of DIP diagnosis. They were the same as for known pregestational type 2 DM. Age was the most important determinant, followed by previous macrosomia (for DIP) and BMI (for PG DM).

Disclosure

M.A. de Campos: None. M.R. Oppermann: None. V.K. Genro: None. C.B. Leitao: None. V.N. Hirakata: None. A.J. Reichelt: None.
1711-P: Dapagliflozin Promotes Adipose Beiging and Lipolysis and Reduces Adipocyte Size in the Obese Prediabetic Dog
Authors: KABIR MORVARID , BERGMAN RICHARD N. , PICCININI FRANCESCA , STEFANOVSKI DARKO , GOPAUL V. SASHI , YANG HSIU-CHIUNG , KOLKA CATHRYN M.
Published: June 2020

Selective sodium glucose cotransporter-2 inhibition promotes urinary glucose excretion, thus reducing blood glucose. Previously we showed that 6 weeks of Dapagliflozin (Dapa) treatment improved insulin sensitivity and prevented weight gain in a canine model of mild type 2 diabetes. Little is known about the effects of Dapa on adipose tissue (AT) function. Eleven dogs were fed a high fat diet for 6 weeks, then received a single low dose of streptozotocin (18.5mg/kg) to induce mild type 2 diabetes. Animals were then exposed to Dapa (n=5, 1.25mg/kg) or placebo (n=6) once per day for 6 weeks, while remaining on the high fat diet. At the end of the study, subcutaneous (SC) and visceral (VIS) fat depots were obtained for adipocytes isolation and molecular studies and fasting plasma parameters were measured. Fasting plasma glucose and insulin levels were unaffected by Dapa treatment. Finite mixture modeling linear regression was used for adipocyte distributions. Dapa decreased adipocyte size and bimodal distributions in both SC and VIS by 10% (P<0.001). Dapa increased UCP1, PGC1α and Prdm16 gene expression by 2.68, 2.34 and 2.16 folds (<0.05) in SC and 2.5, 3.6 and 1.6 folds (<0.05) in VIS depots respectively. Immunostaining of UCP1 in SC and VIS tissues confirmed beiging process. Primary adipocytes were cultured for 4 and 24h. Lipolysis, indicated by glycerol release into the media increased by 40% in SC and 43% in vis after 24h (P<0.05). Adiponectin levels increased only in the SC adipocytes by 80% (P<0.05) after 4 and 24h cultures. Leptin levels increased in SC (by 3 folds P<0.05) and VIS by 2.6 P<0.05) after 24h adipocyte cultures.

In conclusion, Dapa promotes AT beiging, lipolysis, adiponectin and leptin release by adipocytes and decreased adipocyte size and distributions. These changes were independent of glucose and insulin. Thus, for the first time we are beginning to identify mechanisms by which Dapa enhances AT function and improves energy homeostasis in mild type 2 diabetes canine model.

Disclosure

M. Kabir: None. R.N. Bergman: None. F. Piccinini: Employee; Self; Medtronic. D. Stefanovski: None. V. Gopaul: None. H. Yang: Employee; Self; AstraZeneca. C.M. Kolka: Research Support; Self; AstraZeneca.

Funding

AstraZeneca (D1690L00045-CSR212966)
1747-P: MicroRNA-30a Exerts Antifibrotic Effects in Subcutaneous White Adipose Tissue to Preserve Insulin Sensitivity in Obesity
Authors: CHERNIS NATASHA , SAHA PRADIP , FELIX JESSICA B. , MASSCHELIN PETER M. , LIAN LEQIN , COX AARON , KIM KANG HO , WU HUAIZHU , HARTIG SEAN M.
Published: June 2020

White adipose tissue (WAT) is an endocrine organ that dynamically expands and contracts to meet the metabolic demands of the organism. However, prolonged excessive caloric intake overwhelms WAT depots, resulting in peripheral fat deposition, systemic glucose dysregulation, tissue fibrosis, insulin resistance, and type 2 diabetes mellitus. The chronic triggers that degrade WAT function remain undefined, hindering the development of effective strategies to treat obesity and prevent its co-morbidities. We have identified a microRNA, miR-30a, that potently stimulates subcutaneous adipose tissue metabolic flexibility and enhances systemic insulin sensitivity. In follow-up studies, we created a genetically engineered mouse model to conclusively show miR-30a expression in white adipocytes is antidiabetic. Obese mice modified to express miR-30a at elevated levels in adipose tissues maintain insulin sensitivity coupled with reduced hepatic steatosis. Proteomic profiling indicated that miR-30a targets Plasminogen Activator Inhibitor-1 (PAI-1) to limit pro-fibrotic programs that would otherwise restrict subcutaneous WAT expansion and decrease insulin sensitivity. While visceral WAT depots remain a site of significant fibrosis in mice, subcutaneous WAT fibrosis is epidemiologically linked to metabolic disease risk in adult humans. In obese humans, miR-30a expression negatively correlates with PAI-1 levels in subcutaneous WAT, supporting an anti-fibrotic role for miR-30a that ultimately defends insulin sensitivity. Collectively, our observations linking miR-30a expression to insulin sensitivity suggest a new method that restores WAT homeostasis.

Disclosure

N. Chernis: None. P. Saha: None. J.B. Felix: None. P.M. Masschelin: None. L. Lian: None. A. Cox: None. K. Kim: None. H. Wu: None. S.M. Hartig: None.

Funding

American Diabetes Association (1-18-IBS-105 to S.M.H.); National Institutes of Health (R01DK114356)
1766-P: Androgen Action in Hypothalamic Arcuate Nucleus Neurons Increases Hepatic Glucose Production in Female Mice
Authors: MORFORD JAMIE , MAUVAIS-JARVIS FRANCK
Published: June 2020

Androgen excess predisposes females to type 2 diabetes. Using mouse models, we reported that androgen excess causes insulin resistance via activation of the androgen receptor (AR) in the brain. Neurons of the arcuate nucleus of the hypothalamus (ARC) regulate hepatic glucose production (HGP). Thus, we hypothesized that in female mice, androgen excess in neurons of the ARC causes peripheral insulin resistance by increasing HGP. To test this, we injected AaV-Cre-GFP or AaV-GFP into the ARC of ARlox/lox female mice to generate ARC-specific AR knockout (ARC-ARKO) and control mice, respectively. When exposed to a Western diet, control female mice chronically treated with dihydrotestosterone (DHT) developed insulin resistance and fasting hyperglycemia compared to vehicle-treated control mice. In contrast, DHT-treated ARC-ARKO mice remained insulin sensitive and normoglycemic compared to vehicle-treated ARC-ARKO mice. During a hyperinsulinemic-euglycemic clamp, insulin’s ability to suppress HGP was blunted in DHT-treated control mice. In contrast, insulin was still able to suppress HGP in DHT-treated ARC-ARKO females. Additionally, during the clamp, DHT-treated control mice showed no alteration in hepatic activation of AKT, a marker of hepatocyte insulin sensitivity, but exhibited reduced activation of hepatic STAT3, a marker of hypothalamic insulin sensitivity. In contrast, in DHT-treated ARC-ARKO mice activation of hepatic STAT3 was increased. Together these results suggest that in female mice exposed to a Western diet, androgen excess causes insulin resistance in ARC neurons via action at the AR leading to impairments in the brain-IL6-STAT3 pathway which results in unsuppressed HGP.

Disclosure

J. Morford: None. F. Mauvais-Jarvis: None.

Funding

National Institutes of Health (R01DK074970, R01107444); U.S. Department of Veterans Affairs (BX003725)
1768-P: The Role of Hypothalamic Growth Hormone Receptor–Expressing Neurons in Glucose Metabolism and Energy Homeostasis
Authors: LIMA JULIANA , DEBARBA LUCAS K. , AYYAR IVEN T. , UBAH CHIDERA , KHAN MANAL , ALROMDHAN HAWRAA , ALYSOFI MOHAMMED , SADAGURSKI MARIANNA
Published: June 2020

GH and the GH receptor (GHR) are present in regions known to participate in the regulation of feeding, energy balance and glucose metabolism, including the hypothalamus. We have previously demonstrated that GHR signals in nutrient-sensing neurons is critical for hepatic glucose production and lipid metabolism. Given the high expression pattern of GHR in the arcuate nucleus (ARC) of the hypothalamus, we aimed to define the specific role of the ARC GHR-expressing neurons in energy homeostasis and glucose metabolism. For this purpose, we have developed a novel GHR cell-specific molecular tool (GHRcre) using CRISPR/Cas9 gene editing technology. We show that GHR+ neurons in the ARC are mostly co-localized with AgRP, GHRH, and SST neurons and rapidly respond to fasting. Specific chemogenetic activation of GHR+ neurons in the ARC (ARCGHR+) by the DREADDs technique impaired glucose metabolism, suggesting the role of these neurons in metabolic homeostasis. Additionally, while activation of ARCGHR+ neurons increased energy homeostasis (RER, VO2) under metabolic resting phase, the responses to fasting were significantly attenuated, demonstrating the inhibitory role of GHR+ neurons in the ARC for fasting responses. Sirtuin 1 (SIRT1) is highly expressed in the ARC and had been associated with hypothalamic responses to fasting. The majority of ARCGHR+ neurons express SIRT1 and respond to fasting by upregulating the SIRT1 expression. Importantly, hypothalamic upregulation of SIRT1 in response to fasting is blunted in mice lacking GHR in AgRP neurons (AgRPEYFP∆GHR mice), suggesting a novel role of SIRT1 within the hypothalamic GH axis and providing evidence for a link between the somatotropic system and SIRT1 function in responses to fasting.

Disclosure

J. Lima: None. L.K. Debarba: None. I.T. Ayyar: None. C. Ubah: None. M. Khan: None. H. AlRomdhan: None. M. Alysofi: None. M. Sadagurski: None.

Funding

American Diabetes Association (1-18-JDF-063 to M.S.); Wayne State University
178-OR: The Causal Effect of BMI on Health-Related Quality of Life: A Mendelian Randomization Study
Authors: LAXY MICHAEL
Published: June 2020

Background: Previous studies showed that obesity is negatively associated with health related quality of life (HRQoL), however, this association might be biased by reversed causation and omitted variables. To overcome these limitations, this study aims to estimate the causal effect of body mass index (BMI) on HRQoL using genetic variants as instrumental variables - a method that is often called Mendelian Randomization (MR).

Methods: We use a) individual-level data on BMI, 77 identified single nucleotide polymorphism (SNPs), HRQoL measured by the physical (PCS) and mental component score (MCS) of the SF-12 of 3080 participants from a population-based German study, and b) meta-data on the SNP-BMI association of 77 identified SNPs from a meta-analysis with >300,000 individuals. We apply a) an ordinal least square (OLS) regression model, b) a one sample MR approach in which we fit a two stage least square (2SLS) regression model with a genetic SNP-score as instrument, and c) a two sample MR approach in which we estimate Wald estimates using the techniques of inverse variance weighting, simple and weighted medians, and MR Egger regression.

Results: The OLS model shows a 0.32 point decrease in PCS (p<0.001) and a 0.04 point decrease in MCS (p=0.54) per BMI unit. The 2SLS regression of the one sample MR approach indicates endogeneity (Durbin-Wu-Hausman-Test; p<0.05) and shows a nonsignificant 0.92 point decrease in PCS and a nonsignificant 0.32 point decrease in MCS per BMI unit. The two sample MR models also show consistently larger, but non-significant PCS and MCS decreases per BMI unit than the linear model.

Conclusion: The negative effect of BMI on HRQoL might be higher than previous studies suggested. This highlights the potential value resulting from obesity prevention. MR studies with larger sample sizes are needed to obtain more precise causal effect estimates.

Disclosure

M. Laxy: None.
1801-P: Novel De Novo INS P.T97P Variant Presenting with Severe Neonatal DKA
Authors: LAL RAYHAN , MOELLER HANNAH P. , FREEMAN RAQUEL , PRAHALAD PRIYA , ANNES JUSTIN P.
Published: June 2020

Objective: A 5 month old, premature male infant with history of hydrops fetalis and hernia repair presented unresponsive in respiratory distress requiring intubation and was found to have pH 6.93, bicarbonate 6 mmol/L, anion gap 23 mmol/L, glucose 669 mg/dL and HbA1c 14.2%. His condition improved with insulin drip, fluid and electrolyte management. Work-up revealed negative autoantibodies and the University of Chicago Neonatal Diabetes Sequencing Panel was performed. Testing revealed a heterozygous sequence variant of unknown significance in the INS gene (c.289A>C), resulting in the amino acid change p.T97P. In silico pathogenicity prediction tools provided contradictory results. Neither parent carried the variant. INS gene mutations have been associated with Mutant INS-gene-induced Diabetes of Youth (MIDY) that includes permanent neonatal diabetes and MODY 10. Expression of mutant proinsulin is thought to lead to misfolded complexes in the endoplasmic reticulum (ER), leading to ER stress and β-cell death.

Methods: To determine the effects of this variant on insulin processing and release, we utilized a validated luminescent insulin secretion assay where Gaussia luciferase is inserted into the C-peptide portion of proinsulin. We compared luciferase secretion for wild type and variant INS sequences.

Results: The mutation decreased luciferase release ∼10-fold versus wild type. Proteasome inhibition led to partial rescue. However, overall expression of proinsulin-luciferase peptide was unchanged between the wild type and mutant. Therefore, it is likely that the mutation causes disruptions in processes downstream of transcription and translation, such as folding, proteolytic processing and transport.

Conclusions: The novel INS p.T97P is the first identified A-chain mutation resulting in MIDY not involving the creation or deletion of cysteine residues. The substitution of the polar threonine with nonpolar proline may be sufficient to disrupt nearby disulfide bonding in the proinsulin folding pathway.

Disclosure

R. Lal: Consultant; Self; Abbott, Biolinq. H.P. Moeller: None. R. Freeman: None. P. Prahalad: None. J.P. Annes: None.

Funding

National Institutes of Health (1K12DK122550, T32DK007217, R01DK101530, T32GM113854, P30DK116074)
1809-P: Liver Pyruvate Carboxylase Knockout Mice Suggest Noncanonical Sources of Acetyl-CoA for Hepatic Lipid Synthesis
Authors: INIGO MELISSA MAE R. , DEJA STANISLAW , KUCEJOVA BLANKA , FLETCHER JUSTIN A. , FU XIAORONG , BURGESS SHAWN C.
Published: June 2020

Elevated hepatic de novo lipogenesis (DNL) is a key characteristic of patients with Nonalcoholic Fatty Liver Disease (NAFLD). Liver pyruvate carboxylase (PC) may be critical for DNL since PC replenishes oxaloacetate that is converted to citrate as source for acetyl-CoA during DNL. PC may also play a role in triglyceride (TG) synthesis by replenishing oxaloacetate that is used for glyceroneogenesis. Thus, PC is considered a lipogenic enzyme, though most evidence has been derived from non-liver cell culture models and indirect measures in livers of animal models. Liver-specific PC knockout mice (LPCKO) have lower circulating TG on a high fat diet, but fasting liver TG was not improved. To determine whether liver-specific PC is required for DNL and glyceroneogenesis, we examined LPCKO mice using a 1H/2H NMR approach to directly measure flux through DNL and TG-glycerol synthesis. PCf/f and LPCKO mice were exposed to either standard chow or a high sucrose diet (HSD) for 10 days. Ad libitum fed PCf/f and LPCKO mice were then administered D2O overnight. Liver lipids were extracted, and positional isotopomer analysis by 1H/2H NMR was used to determine DNL and TG-glycerol synthesis fluxes. DNL flux was not significantly different between PCf/f and LPCKO mice under standard chow conditions and only tended to be moderately lower in LPCKO mice after a short-term HSD treatment. TG-glycerol synthesis was also similar between PCf/f and LPCKO, regardless of diet. Surprisingly, liver triglyceride levels were modestly higher in standard chow fed LPCKO than PCf/f mice, and no significant differences were found between HSD-fed PCf/f and LPCKO mice. Our initial findings suggest that PC is not required for DNL and TG-glycerol synthesis during the fed state and therefore indicate that alternative pathways may exist to supply of cytosolic acetyl-CoA for lipid synthesis.

Disclosure

M.R. Inigo: None. S. Deja: None. B. Kucejova: None. J.A. Fletcher: None. X. Fu: None. S.C. Burgess: None.

Funding

Robert A. Welch Foundation (I-1804)
1873-P: Glycemic Impact of Pancreatic Enzyme Replacement in Patients with Diabetes and Exocrine Pancreatic Insufficiency: A Case Series
Authors: UWAIFO GABRIEL I.
Published: June 2020

Some patients with type 2 diabetes (T2DM) and/or pancreatogenous diabetes have exocrine pancreatic insufficiency (EPI). The impact of pancreatic enzyme replacement (PER) on diabetes (DM) control is largely undocumented and unknown. We present 4 cases suggesting PER may positively impact DM glycemic and lipids. Case 1 is a 46 yr Caucasian (C) man with T2DM and recurrent gall stone pancreatitis. Prior to onset his HBA1c was between 6.3-7.4 but with recurrent episodes reached peak of 10.8. He was diagnosed with chronic pancreatitis with EPI and commenced PER. After 3 mths on PER his HBA1c was 6.7 with resolution of hypertriglyceridemia (HTG) and improved total cholesterol (TC). Case 2 is a 57 yr old C man with T2DM and episodic HTG related pancreatitis. He had erratic medication access due to insurance limitations. He was recently diagnosed with EPI but initially could not afford PER. HBA1c at diagnosis of EPI was 9.7 with elevated TC and HTG. When he secured PER, within 8 wks of use HBA1c reduced to 7.6 with improved HTG and TC. Case 3 is a 24 yr old C man with known cystic fibrosis and cystic fibrosis related DM. Initial HBA1c was 8.6 and he had been of PER voluntarily for yrs. He had features of EPI and was urged to resume PER which he did. At 4 mth follow up after restarting PER his HBA1c was 6.4 with improvement in HTG. Case 4 is a 36 yr old C man with prior well controlled T2DM and known MEN-1. His prior mean HBA1c was 6.7. Due to recurrent bouts of severe hypoglycemia in the prior 2 yrs work up revealed multiple pancreatic insulinomas for which he had partial pancreatectomy. His DM control declined with peak HBA1c of 11.3. He developed symptoms of EPI and was commenced on PER. 3mths after starting PER his HBA1c was 7.1 with resolution of HTG and improved TC. Early recognition of EPI in patients with DM is important. Commencement of PER in such patients may positively impact glycemia and dyslipidemia especially HTG. Further study of the relationship of PER to DM metabolic control is warranted.

Disclosure

G.I. Uwaifo: None.
190-OR: Impact of Prenatal Exposure to Maternal Obesity and Child Physical Activity Levels on Neurocognition
Authors: ALVES JASMIN , ANGELO BRENDAN C. , LUO SHAN H. , CHOW TING , YUNKER ALEXANDRA G. , XIANG ANNY , PAGE KATHLEEN A. , ZINK JENNIFER , BELCHER BRITNI R. , CLARK KRISTI
Published: June 2020

Prenatal exposure to maternal obesity is linked to neurocognitive deficits in offspring. In contrast, engaging in physical activity (PA) is beneficial for cognition and brain development. We aimed to determine if greater PA is associated with higher intelligence quotient (IQ) scores and greater brain white matter integrity in children born to mothers with a range of pre-pregnancy BMI. Ninety-nine children (59 girls) ages 7-13 years completed the study. Maternal pre-pregnancy BMI was obtained from electronic medical records. Wechsler Abbreviated Scale of Intelligence was used to measure IQ. Diffusion tensor imaging was used to measure mean global fractional anisotropy (FA), a marker of white matter integrity. Time spent in moderate-to-vigorous PA (MVPA) and vigorous PA (VPA) was assessed using 3-day PA recalls. Linear regressions were used to assess whether maternal pre-pregnancy BMI or child MVPA and VPA were associated with child IQ scores and global FA. Maternal pre-pregnancy BMI ranged from 19.0-54.0 kg/m2 (43% obese). Mean±SD child IQ was 106.4±14.0, MVPA was 132.8±95.4 min/day, and VPA was 19.0±27.1 min/day. Maternal pre-pregnancy BMI was negatively associated with child IQ scores (β=-0.31; p=0.10); this marginal association remained after adjusting for child age, sex, puberty, BMI z-score, and socioeconomic status (β=-0.42; p=0.095). MVPA was not associated with child IQ (β=2.63; p=0.16). However, children who spent more time in VPA had higher IQ scores before (β=7.05; p=0.003) and after adjusting for the same co-variates (β=5.65; p=0.024) and further adjusting for maternal pre-pregnancy BMI (β=5.01; p=0.049). Children who spent more time in VPA also had greater mean global FA (unadjusted: β=0.01; p=0.017; adjusted: β=0.01; p=0.005). These findings suggest that engaging in VPA is beneficial for cognition and white matter microstructure in children, independent of maternal pre-pregnancy BMI.

Disclosure

J. Alves: None. B.C. Angelo: None. S.H. Luo: None. T. Chow: None. A.G. Yunker: None. A. Xiang: None. K.A. Page: None. J. Zink: None. B.R. Belcher: None. K. Clark: None.

Funding

American Diabetes Association (1-14-ACE-36 to K.A.P.); National Institute of Diabetes and Digestive and Kidney Diseases (R01DK116858); National Institute of Mental Health (F31MH115640)