bluuebunny/crossref_metadata_2025_split huggingface dataset:
193-OR: Physiology-Informed Type 2 Diabetes Genetic Clusters and Gestational Diabetes
Authors: POWE CAMILLE E. , UDLER MIRIAM , HSU SARAH , KUANG ALAN , ALLARD CATHERINE , MANNING ALISA , BOUCHARD LUIGI , PERRON PATRICE , FLOREZ JOSE C. , LOWE WILLIAM , SCHOLTENS DENISE , HIVERT MARIE-FRANCE 
Published: June 2020

Background: We previously described physiology-informed genetically-anchored type 2 diabetes (T2D) clusters based on associations between glycemic traits and T2D-associated variants discovered in prior GWAS. Cluster polygenic scores (PSs) delineate T2D subtypes with distinct features (e.g., reduced beta-cell function, obesity, lipodystrophy-like). These PSs have not been tested for association with gestational diabetes (GDM).
Methods: We tested for associations between PSs (beta-cell, proinsulin, lipodystrophy, obesity, liver/lipid) and traits at 24-30 weeks gestation in the Gen3G cohort (N=582, 97% white non-Hispanic) with linear regression (β=change per 1 SD increase in PS). In 3 cohorts [Gen3G, HAPO (N=1369, 100% white non-Hispanic) MGH2(N=621, 62% white non-Hispanic)], we tested for associations between PSs and GDM using logistic regression and conducted fixed effects meta-analyses.
Results: The beta-cell PS was associated with higher 1-hr OGTT glucose (β=4.5 mg/dl, P&lt;0.001) and lower insulin secretory response (β=-45.7 Stumvoll 1st phase index, P=0.02). The obesity PS was associated with higher BMI (β=0.5 kg/m2, P=0.03) and HbA1c (β=0.03%, P=0.02). The lipodystrophy PS was associated with higher 1-hr OGTT glucose (β=2.6 mg/dl, P=0.03), higher fasting triglycerides (TGs) (β=6 mg/dl, P=0.01), and lower insulin sensitivity (β=-0.6 Matsuda index, P=0.005). The liver/lipid PS was associated with lower fasting TGs (β=-11 mg/dl, P&lt;0.001). In 3-cohort meta-analyses (297 cases), only the beta-cell PS was associated with GDM (OR 1.19, P=0.009). There was heterogeneity across cohorts for the liver/lipid PS-GDM association. No PS was associated with birth weight. Adjustment for maternal age and genetic principal components did not alter results.
Conclusion: Clusters of T2D-associated genetic variants derived based on physiology outside of pregnancy are associated with expected traits in pregnancy. A cluster representing reduced beta-cell function is associated with GDM.

Disclosure
C.E. Powe: None. M. Udler: None. S. Hsu: None. A. Kuang: None. C. Allard: None. A. Manning: None. L. Bouchard: None. P. Perron: None. J.C. Florez: Advisory Panel; Self; Doris Duke Charitable Foundation. Other Relationship; Self; Novo Nordisk Inc., Park Street School. W. Lowe: None. D. Scholtens: None. M. Hivert: None.

Funding
American Diabetes Association (1-15-ACE-26 to M-F.H.); National Institutes of Health (K23DK113218); Robert Wood Johnson Foundation; Harold Amos Medical Faculty Development Award (72456)
1930-P: The Knockdown of TRIL in POMC-Expressing Neurons Reduces Body Adiposity and Ameliorates Leptin Responsiveness
Authors: MOURA-ASSIS ALEXANDRE , JUNIOR JOSÉ , GASPAR JOANA M. , VELLOSO LICIO , NOGUEIRA PEDRO AUGUSTO S. 
Published: June 2020

Chronic HFD intake induces low-grade inflammation in the hypothalamus, which is characterized by proinflammatory cytokine overproduction. This process occurs, at least in part, via TLR4 activation by circulant saturated fatty acids. The TLR4 signaling in the brain is mediated by the TLR4 interactor with leucine-rich repeats (TRIL), an upstream accessory protein. Our main purpose was to determine the putative involvement of TRIL in diet-induced hypothalamic inflammation. We employed male, 8-12 week-old C57BL and POMCCRE mice fed on chow or HFD. The mRNA levels in the hypothalamus were measured by RT-PCR, western blot analyses was used for determination of protein content and immunofluorescence microscopy for evaluate the distribution of TRIL in arcuate nucleus (ARC) of hypothalamus and leptin signaling in POMC-expressing neurons. Finally, bilateral viral injections were performed to non-specific knockdown of TRIL in ARC using shRNA-based lentivirus and Cre-dependent AAV to knockdown TRIL specifically in POMC neurons. The non-specific knockdown of TRIL in the ARC of mice fed on HFD attenuated body weight gain despite no alterations in food intake, improved glucose homeostasis and increased energy expenditure. Moreover, nlrp3, il1β and hspa5 mRNA expression were decreased upon knockdown of TRIL. POMC-specific knockdown of TRIL reduced body fat with no changes in food intake or body weight gain in mice fed on HFD for 8 weeks. The inhibition of TRIL in POMC neurons resulted in a trend of improved leptin signaling in those neurons. We also found increased expression of thermogenic genes in the iBAT and improved fasting glucose. POMC-specific inhibition of TRIL in mice fed on HFD for long-term (24 weeks) did not prevent POMC loss or diminishes cleaved-caspase 3 in ARC. Collectively our results suggest that TRIL is involved in the TLR4-mediated early response to dietary fats and TRIL targeting counteract hypothalamic inflammation and partially restores metabolic homeostasis.

Disclosure
A. Moura-Assis: None. J. Junior: None. J.M. Gaspar: None. L. Velloso: None. P.S. Nogueira: None.

Funding
São Paulo Research Foundation (2016/01245-5)
1987-P: Inflammation and Insulin Resistance in Adipose Tissue
Authors: SØNDERGAARD ESBEN , ESPINOSA DE YCAZA ANA E. , MORGAN-BATHKE MARIA , LYTLE KELLI , DELIVANIS DANAE A. , CARRANZA LEON BARBARA G. , JENSEN MICHAEL D. 
Published: June 2020

Background: Adipose tissue inflammation is linked to adipose tissue insulin resistance. However, whether this is a causal relationship remains controversial. Therefore, we aimed to explore the relationship between measures of adipose tissue inflammation and adipose tissue insulin sensitivity over a wide range of body fat distribution and adipose tissue insulin sensitivity.
Methods: We recruited a total of 87 volunteers (30 men and 57 women). Adipose tissue insulin sensitivity was quantified as the insulin concentration required for a 50% suppression of lipolysis (IC50). Lipolysis was measured as steady-state palmitate flux before and during a hyperinsulinemic-euglycemic clamp. Adipose tissue macrophage (ATM) was quantified by immunohistochemistry with antibodies against CD68 (total ATM), CD14 (pro-inflammatory ATM) and CD206 (anti-inflammatory ATM) and senescent preadipocyte content was quantified by β-galactosidase positivity using fluorescent microscopy in abdominal and femoral fat biopsies.
Results: Total ATM content was only weakly associated with IC50 in abdominal fat (r=0.26, p=0.01) and no association was observed for femoral fat. No association was observed between IC50 and pro-inflammatory ATM content. Anti-inflammatory ATM content was associated with IC50 in abdominal (r=0.46, p&lt;0.001) and femoral (r=0.33, p=0.02) fat. Senescent preadipocyte content was weakly associated with IC50 in femoral fat (r=0.36, p=0.01), but no association was observed in abdominal fat. Fat cell size was strongly associated with IC50 in both abdominal (r=0.6 p&lt;0.001) and femoral (r=0.48, p&lt;0.01) fat.
Conclusions: ATM content is only weakly associated to adipose tissue insulin sensitivity. Only the content of anti-inflammatory, not pro-inflammatory macrophages, was associated with adipose tissue insulin sensitivity. This argues against pro-inflammatory macrophages as mediators of insulin resistance in adipose tissue.

Disclosure
E. Søndergaard: None. A.E. Espinosa De Ycaza: None. M. Morgan-Bathke: None. K. Lytle: None. D.A. Delivanis: None. B.G. Carranza Leon: None. M.D. Jensen: None.

Funding
National Institutes of Health (DK40404, DK45343)
200-LB: Ceramide Kinase CERK Regulates Inflammatory Responses in Monocytes/Macrophages Induced by TNF-a
Authors: ALRASHED FATEMA , AHMAD RASHEED 
Published: June 2020

Obesity is associated with elevated levels of TNF-α and proinflammatory CD11c monocytes/macrophages. TNF-α mediated dysregulation in the plasticity of monocytes/macrophages is concomitant with pathogenesis of several inflammatory diseases, including metabolic syndrome, but the underlying mechanisms are incompletely understood. Since Ceramide kinase (CERK; a critical mediator of eicosanoid synthesis, and its product, ceramide-1-phosphate (C1P)) is a key enzyme for ceramide production involved in inflammation, we investigated whether the CERK contributed to the inflammatory changes in the monocytes/macrophages induced by TNF-α. In this study, we demonstrate that the disruption of CERK activity in monocytes/macrophages either by chemical inhibitor NVP- 231 or small interfering RNA (siRNA)against CERK gene results in defects in the TNF-α mediated expression of CD11c and CD11b and HLA-Dr. Furthermore, blockage of CERK in monocytes/macrophages inhibited the secretion of IL-1b and MCP-1. Phosphorylation of JNK, p38 and NF-κB resulting from TNF-α stimulation was also attenuated by the inhibition of CERK. Moreover, NF-kB/AP-1 activity was blocked by the inhibition of CERK. Our human data show that CERK was elevated in PBMCs from obese individuals and positively corelated with TNF-α. These findings indicate that CERK acts, in a part, as a master switch in the TNF-α mediated inflammatory responses in monocytes/macrophages.

Disclosure
F. Alrashed: None. R. Ahmad: None.

Funding
Kuwait Foundation for the Advancement of Sciences (RA-AM 2016007)
2021-P: Comparing BMI and Physical Activity with Diabetes Risk in College Freshman: The HERD Study
Authors: POWELL ROBERT , CYPHERT HOLLY 
Published: June 2020

Obesity and Physical Inactivity are leading risk factors for insulin resistance and type 2 diabetes. West Virginia leads the U.S. in obesity rates and ranks 11th for no leisure time physical activity. Data is scarce examining modifiable risk factors in young adults transitioning from high school to college. The purpose of this analysis was to examine the relationship between BMI and Physical Activity on insulin levels in college-aged freshman enrolled in the HERD (Higher Education Reducing Diabetes) Study at Marshall University in Huntington, West Virginia. The HERD Study is a 4 year, randomized study examining the effects of a freshman healthy lifestyle intervention on type 2 diabetes risk. One hundred eighty three freshman enrolled into the study [age= 18.3±1.5 years; female=59.4%(n=107); 85.5% Caucasian(n=153); BMI= 27.3±8.1 kg/m2; VO2max=35.6 ±8.59 ml/kg/min]. Diabetes risk values were A1c= 5.09±.43%; fasting glucose= 85.9±9.66 mg/dl; insulin= 0.76±.60ng/ml. Fifty four percent (n=92) of students were deemed overweight or obese (BMI ≥ 25 kg/m2) while 67% (n= 122) did not meet current physical activity guidelines. No significant differences were found between BMI and A1c (p=.48) or fasting glucose (p=.82). However, students with a BMI ≥25 kg/m2 showed fasting insulin levels twice as high as students with a BMI &lt;25 kg/m2 (.50 vs. .98 ng/ml, p=.003). With physical activity, there was a significant, inverse correlation between VO2max and insulin levels (r=-.23, p=.048). However, no differences were found between students whom aerobic train vs. not aerobic train (p=.60). A trend was shown where resistance trained students had lower insulin levels compared to non-resistance trained (.52+.61 vs. .77+.37 ng/ml, p=.06). BMI and fitness measures appear to impact insulin levels in college-aged freshman thereby increasing their risk for diabetes. Future analysis will examine the effects of lifestyle intervention on improvements in BMI and cardiometabolic markers.

Disclosure
R. Powell: None. H. Cyphert: None.
2030-P: The Efficacy of Laparoscopic Sleeve Gastrectomy in Severely Obese Patients with Varying Serum Triglyceride Levels
Authors: LI GUIFANG , HUANG YUEYE , ZHANG JUNYI , WANG XINGCHUN , QIAN CHUNHUA , CHENG XIAOYUN , QU SHEN 
Published: June 2020

Purpose: To investigate the metabolic improvements in obese patients with different serum triglyceride (TG) levels after laparoscopic sleeve gastrectomy (LSG).
Methods: 118 severely obese patients [body mass index (BMI)&gt;35kg/m2] underwent LSG with complete information of follow-up for 3 and 6 months were analyzed. Patients were divided into two groups based on their different baseline serum levels of TG.
Results: The metabolic parameters (BMI, NC, WC, HC, WHR, blood pressure, liver function, blood glucose, blood lipids, insulin resistance, HbA1c, and UA) of the normal and higher TG groups were significantly improved after surgery. Patients with normal TG had better improvement in BMI and WC compared with higher TG patients in both 3 and 6 months. After adjustment for age and gender, ΔWC positively correlated with ΔBMI (r=0.446, P&lt;0.001), creatinine level (ΔCr, r=0.365, P=0.005) in the normal group in 3 months follow-up. In 6 months after LSG, ΔWC positively correlated with ΔBMI (r=0.657, P&lt;0.001) and ΔCr (r=0.442, P=0.009) while ΔWC negatively correlated with the change in eGFR level (ΔeGFR, r=-0.374, P=0.009) in the normal group. In addition, multivariate linear regression analysis showed that decreased ΔWC levels significantly correlated with ΔBMI (β=1.333, R2 =0.259, P&lt;0.001) and ΔCr (β=0.336, R2 =0.160, P=0.005) both in 3 months and 6 months after LSG.
Conclusion: LSG can effectively improve metabolic disorders. Moreover, there were differences in weight loss between the normal and high TG groups after LSG, indicating that the baseline TG level can be used to predict short-term weight loss after LSG.

Disclosure
G. Li: None. Y. Huang: None. J. Zhang: None. X. Wang: None. C. Qian: None. X. Cheng: None. S. Qu: None.

Funding
National Key Research and Development Program of China (2018YFC1314100); National Natural Science Foundation of China (81970677); Shanghai Pujiang Program (2019PJD040); Shanghai Committee of Science and Technology (18411951803, 17DZ1910603)
2043-P: Hyaluronan Produced by Inflamed Islet Endothelial Cells Contributes to HIAPP Aggregation
Authors: CASTILLO JOSEPH J. , HOGAN MEGHAN F. , APLIN ALFRED , ESSER NATHALIE , TEMPLIN ANDREW T. , HULL REBECCA L. 
Published: June 2020

In type 2 diabetes (T2D), aggregation of human islet amyloid polypeptide (hIAPP) occurs within the islet extracellular matrix (ECM) and is toxic to β cells. We recently showed that hIAPP aggregation also induces a pro-inflammatory response in islet endothelial cells (ECs). In most tissues, this inflammatory response in ECs is mediated by upregulating production of ECM components, such as hyaluronan (HA). Indeed, we find that HA accumulates in the islet ECM of hIAPP transgenic mice, where it was colocalized with amyloid deposits, while no HA accumulation occurred in non-transgenic littermate controls (which do not develop islet amyloid). Therefore, we hypothesized that hIAPP aggregation results in HA production from islet ECs, and that this HA production exacerbates hIAPP deposition/toxicity. To test this hypothesis, we utilized immortalized murine islet ECs (MS-1 cells), which under normal conditions express low levels of HA synthase (Has isoforms 2 and 3). Treatment of these cells for 48 h with hIAPP (20 µM) increased Has2 and Has3 mRNA levels (23±4 and 8±1 fold over vehicle; n=4, p&lt;0.05 for both), which was abrogated upon co-incubation with Congo red, an inhibitor of hIAPP aggregation (1.4±0.3 and 1.4±0.2 for Has2 and Has3, respectively; n=4). Next, we used a thioflavin T assay, wherein fluorescence is proportional to fibril formation from amyloidogenic peptides, to determine the kinetics of hIAPP aggregation ± HA. We found co-incubation of HA with synthetic hIAPP peptide resulted in a marked acceleration (log time of 1 vs. 40 hours) and increase in maximal (greater than 2-fold) hIAPP accretion. In sum, exposure of islet ECs to aggregated hIAPP increases HA synthesis by these cells, which in turn further exacerbates aggregation of hIAPP to form amyloid. Thus, interventions targeting endothelial cell inflammation and/or HA production may be useful in blocking amyloid formation and its toxic effects in T2D.

Disclosure
J.J. Castillo: None. M.F. Hogan: None. A. Aplin: None. N. Esser: None. A.T. Templin: None. R.L. Hull: None.

Funding
National Institutes of Health (T32HL007028)
2049-P: The Chd4 Helicase of the Nucleosome Remodeling and Deacetylase Complex Modulates Pdx1 Target Gene Expression In Vitro
Authors: DAVIDSON REBECCA K. , CASEY NOLAN , SPAETH JASON 
Published: June 2020

During the progression of type 2 diabetes, β-cells fail and a subset lose features associated with their identity including reduced expression of key transcription factors (TFs). Among these is Pdx1, which is necessary for maintaining β-cell function and identity by controlling the expression of numerous β-cell genes including insulin, MafA, Glut2, and Ucn3. Pdx1, like most TFs, depends on the recruitment of coregulators to modulate its transcriptional activity. Numerous Pdx1-interacting proteins have been identified including Chd4, an ATP-dependent helicase of the chromatin remodeling NuRD complex. NuRD controls cellular processes including maintenance of cellular identity and mitochondrial function in a variety of tissues. The role Chd4 plays in modulating Pdx1 activity in the β-cell has yet to be explored. In this study, we performed co-immunoprecipitation and proximity ligation assay (PLA) in the Min6 murine insulinoma β-cell line and PLA in 8-week old C57BL/6 mouse pancreatic tissue, confirming Chd4:Pdx1 interactions in the nuclei of β-cells in both cell lines and tissue. Transient, RNAi-mediated knockdown of Chd4 in mouse β-cell lines altered the expression of Pdx1-regulated genes involved in β-cell function and identity including the MafA TF, which was reduced at both the RNA (41% ± 3 (p &lt; 0.05)) and protein level. Moreover, the nuclear-encoded mitochondrial Tfam TF (reduced 30% ± 10 (p = 0.09)), the mitochondrial Mipep enzyme (reduced 35% ± 18 (p = 0.10)), and pre-Ins2, the unprocessed Ins2 transcript (reduced 38% ± 9 (p = 0.13)) were also altered, although they did not reach significance. These data suggest that Chd4 plays an important role in modulating the expression of genes controlled by Pdx1 in the β-cell. Future efforts are focused on further characterizing the role of Chd4 in the β-cell with CRISPR/Cas9 Chd4 knockout β-cell lines in vitro and β-cell specific Chd4 knockout mice.

Disclosure
R.K. Davidson: None. N. Casey: None. J. Spaeth: None.

Funding
National Institute of Diabetes and Digestive and Kidney Diseases (K01DK115633)
2067-P: Cell Therapies for Diabetes Using Spontaneous Canine Diabetes as a Translational Model
Authors: WILDEY ALDYN , WILLIAMS SUSAN J. , HAMILTON MEGAN , HARRINGTON STEPHEN , OTT LINDSEY , KARANU FRANCIS 
Published: June 2020

FDA approved Clinical Trials are already under way using stem cell-derived insulin-producing cells (IPCs) for the treatment of type 1 diabetes in humans. Differentiation methods used to derive beta-cell like IPCs have greatly improved over the last few years, resulting in highly efficacious treatment of high blood glucose in diabetic animals models. However, there still remains major challenges of developing an effective cure in humans due to challenges of delivery and long term survival and function of grafts. Like humans, spontaneous canine diabetes is a commonly diagnosed, multifactorial endocrine disorder in dogs which clinically presents as a type 1 diabetes disorder. In order to develop successful human diabetes therapies using stem cells, spontaneous canine diabetes offers a very attractive translational model. Dogs have major similarities to humans including the clinical signs, pathophysiology and long-term clinical management of the disease by use of exogenous insulin injections. Using cadaveric canine islets and our Core-Shell Spherification (CSS) microencapsulation methods, we have successfully reversed hyperglycemia in dogs for varying time periods. Our data confirms that similar to humans, islet transplantation is a viable option for diabetes treatment in dogs. To further develop canine diabetes as a translational model for human diabetes, while developing stem cell therapies for dogs, we have successfully generated IPCs from human pluripotent stem cells. The IPC clusters express key beta-cell biomarkers including PDX1, NK6.1, NeuroD1, and insulin. The majority of endocrine cells in clusters are single hormonal where separate cells express ARX and glucagon. These cell clusters corrected induced diabetes in mouse models when transplanted under the kidney capsule. Our studies will further allow us to further test the use of the canine model, both in allo- and xenogeneic transplant environments, for further pre-clinical development in humans.

Disclosure
A. Wildey: None. S.J. Williams: None. M. Hamilton: None. S. Harrington: None. L. Ott: Employee; Self; Likarda, LLC. F. Karanu: Employee; Self; Likarda, LLC.
2076-P: Increased Gap Junction Coupling in the Islet of Langerhans following Caloric Restriction
Authors: AMARAL MARIA C. , KRAVETS VIRA , DWULET JAEANN M. , FARNSWORTH NIKKI L. , PISCOPIO ROBERT A. , SCHLEICHER WOLFGANG , MIRANDA JOSE , BENNINGER RICHARD K. 
Published: June 2020

Intercellular communication mediated by connexin 36 gap junction channels coordinate and regulate the dynamics of islet calcium oscillations and insulin secretion. Impaired gap junction coupling occurs in prediabetic mice under a high fat diet. Dietary interventions, such as calorie restriction, can regain weight and modulate insulin secretion. However, it is unknown whether caloric restriction can recover impaired gap junctions coupling and calcium regulation in islets of mice fed a high fat diet. Here, we tested if caloric restriction can protect against obesity-induced decreases in connexin36 coupling and altered islet function. We used C57BL6 mice under a normal diet (Envigo 2020X), or a high fat diet (HFD) (Envigo TD.03584) which causes prediabetes after three months. HFD mice were also submitted to 40% caloric restriction (2g/day) for one month. We performed intraperitoneal glucose tolerance tests, insulin tolerance tests, in vivo insulin level measurements. In isolated islets we measured glucose-stimulated insulin secretion, islet Ca2+ dynamics, metabolic activity, along with gap junction permeability. We observed that caloric restriction improved glucose tolerance (p=0.002), insulin sensitivity (p=0.001), and islet function. This included normalization glucose-induced insulin secretion (p=0.01), enhanced Ca2+ oscillation amplitude (p=0.002) and greater coordination (p=0.04). These improvements correlated with restoration in gap junction permeability (p=0.02). Caloric restriction also promoted a slight increase in NAD(P)H increase by elevated glucose (p=0.04) indicating improved metabolic activity and viability of these cells. We conclude that the decline in gap junction coupling. mediated by connexin 36 in islets that occurs in diet-induced obesity, is improved by short term caloric restriction. This improvement contributes to improved islet function.

Disclosure
M.C. Amaral: None. V. Kravets: None. J.M. Dwulet: None. N.L. Farnsworth: None. R.A. Piscopio: None. W. Schleicher: None. J. Miranda: None. R.K. Benninger: None.

Funding
JDRF (5-CDA-2014-198-A-N); National Institutes of Health (R01DK102950, R01DK106412 to R.K.B.); Fundação de Amparo à Pesquisa do Estado de São Paulo (2018/04536-6)