bluuebunny/crossref_metadata_2025_split huggingface dataset:
2310-PUB: Chronic Western Diets Initiate Fibrosis Progression in Nonalcoholic Fatty Liver of Male Mice Modifying the PPARγ Pathway via Mir-27b-5p Regulation
Authors: ZHANG JIAN , POWELL CATHERINE , KAY MATTHEW K. , SONKAR RAVI , MERUVU SUNITHA , CHOUDHURY MAHUA 
Published: June 2020

Increased fructose intake in Western diets contributes to metabolic diseases. However, the epigenetic mechanisms are mostly unknown. Here, six week-old C57BL/6J male and female mice were fed with a low-fat diet (LFD), high-fat diet (HFD), and high-fat high-fructose diet (HFD-HF) for 20 weeks. We determined that HFD-HF or HFD mice experienced significant metabolic dysregulation compared to the LFD. Metabolic changes in male mice were significantly dysregulated compared to female mice. HFD-HF and HDF male-specific responses included physiological/metabolic changes such as significant increased body weight, increased liver size, increased fasting glucose levels and downregulated PPARγ (P &lt; 0.001), SCD1 (P &lt; 0.05 to P &lt; 0.01), and FAS (P &lt; 0.05) protein expression. In contrast, female mice were less affected by HFD and HFD-HF. Interestingly, none of the groups exhibited less insulin sensitivity after 20 weeks of feeding when compared to LFD. As miR-27b contains a seed sequence in PPARγ, it was discovered that these changes are accompanied by male-specific upregulation of miR-27b-5p, which is even more pronounced in the HFD-HF group (P &lt; 0.01 vs. LFD) compared to the HFD group (P &lt; 0.05 vs. LFD). Other miR-27 subtypes were increased but not significantly. Fat ballooning in hepatocytes were increased in HFD fed mice compared to HFD-HF fed mice, however, the HFD-HF liver showed an increase in the number of small cells. Here, we concluded that our chronic western diet-composition administered for 20 weeks surpassed the nonalcoholic fatty liver (NAFL) stage and initiated liver fibrosis via miR-27b-5p-induced PPARγ downregulation.

Disclosure
J. Zhang: None. C. Powell: None. M.K. Kay: None. R. Sonkar: None. S. Meruvu: None. M. Choudhury: None.

Funding
Morris L. Lichtenstein, Jr. Medical Research Foundation
2313-PUB: Alterations in Basal State of Insulin and mTOR-Dependent Signalings Closely Related to Impaired Incretin Profile and Type 2 Diabetes in Subcutaneous Adipose Tissue of Obese Patients
Authors: STAFEEV IURII , SKLYANIK IGOR , MICHURINA SVETLANA , SHESTAKOVA EKATERINA , YURASOV ANATOLIY , YAHYAEV KAMIL , KARMADONOV ANDREY , RATNER ELIZAVETA , MENSHIKOV MIKHAIL , VOROTNIKOV ALEXANDER V. , PARFYONOVA YELENA V. , SHESTAKOVA MARINA V. 
Published: June 2020

Objective: In our study we have compared basal insulin and mTOR signaling in subcutaneous fat of obese T2DM vs. obese subjects with normal glucose tolerance (NGT). We have performed correlation analysis of phosphorylation’s parameters against clinical parameters of carbohydrate metabolism and incretin secretion profiles.
Methods: 22 patients with long (&gt;10 years) and morbid (BMI &gt; 35 kg/m2) obesity, 12 of which had NGT and 10 had T2DM were enrolled in this study. Hyperinsulinemic-euglycemic clamp test and HOMA-IR were used to measure insulin resistance. Blood samples taken at 0, 30 and 120 min of food load test were used to assess incretin profile, insulin and glucose levels. Amount of total and visceral fat was determined by bioelectrical impedance analysis. Subcutaneous fat biopsies were obtained during bariatric surgery for all patients and signaling state was analyzed by western blots.
Results: As assessed by western blots of insulin receptor substrate (IRS), Akt, Raptor, Rictor, mTOR and S6K1, the basal insulin signaling and mTORC activities were comparable in NGT and T2DM groups, whereas phosphorylation of AS160 (pAS160-S318) was significantly lower and serum and glucocorticoid-induced kinase (SGK; pSGK-S422) was significantly higher in T2DM group. Various correlations were found between the degree of insulin resistance and amount of visceral fat, changes in incretin profile, glucose metabolic parameters and phosphorylation level of AS160, incretin secretion profile and phosphorylated levels of AS160 or SGK1.
Conclusions: Altered phosphorylation of AS160 and SGK1 is associated with obese T2DM phenotype and impaired incretin profile. We think that AS160 and SGK1 phosphorylations can be important markers of the prediabetes-to-diabetes transition.

Disclosure
I. Stafeev: None. I. Sklyanik: None. S. Michurina: None. E. Shestakova: Employee; Spouse/Partner; AstraZeneca. Speaker’s Bureau; Self; Abbott, AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Novartis Pharma K.K., Novo Nordisk A/S, Sanofi-Aventis, Takeda Pharmaceutical Company Limited. A. Yurasov: None. K. Yahyaev: None. A. Karmadonov: None. E. Ratner: None. M. Menshikov: None. A.V. Vorotnikov: None. Y.V. Parfyonova: None. M.V. Shestakova: None.

Funding
Russian Science Foundation (17-15-01435)
2315-PUB: High Serum Ferritin Predicts Inferior Diabetes-Related Outcomes after Gastric Bypass but Not Sleeve Gastrectomy Bariatric Surgery
Authors: TONG JENNY , ALVAREZ RAFAEL , RUSSELL GREGORY B. , KHOURI ALEXANDER N. , SEELEY RANDY J. , MCCLAIN DONALD A. 
Published: June 2020

Objective: Because higher iron level is associated with increased risk of type 2 diabetes and bariatric surgery is known to alter iron homeostasis, we examined the effects of tissue iron stores on glycemic outcome after bariatric surgery.
Research Design and Methods: We performed a retrospective analysis of glycemia outcomes after Roux-en Y gastric bypass (RYGB) or vertical sleeve gastrectomy (VSG) in individuals with elevated hemoglobin A1c (HbA1c) as a function of serum ferritin before and after surgery, using two large and independent electronic health databases from the Carolinas Collaborative and University of Michigan.
Results: Improvement of HbA1c, weight, and alanine aminotransferase (ALT) was comparable between the two procedures 6-12 months after surgery. For gastric bypass procedures, however, the improvement in HbA1c and the chance of normalizing HbA1c were inversely proportional to preoperative ferritin, while no such relationship existed for gastrectomy. The improvement in ALT was correlated with the decrease in serum ferritin after surgery for both surgery types. Changes in hemoglobin were also related to preoperative ferritin.
Conclusion: Preoperative iron stores may modulate glycemic improvement after bariatric surgery differentially depending on procedure type. This has implications regarding the operation of choice in certain patient populations.

Disclosure
J. Tong: None. R. Alvarez: None. G.B. Russell: None. A.N. Khouri: None. R.J. Seeley: Consultant; Self; Ionis Pharmaceuticals, Inc., Kintai, Kintai, Novo Nordisk A/S, Sanofi, Scohia Pharma Inc. Research Support; Self; AstraZeneca, Novo Nordisk A/S, Pfizer Inc. Stock/Shareholder; Self; Redesign Health. D.A. McClain: None.

Funding
National Institutes of Health (R01DK097550)
2316-PUB: Lipidomic Profiling Reveals Distinct Differences in Plasma Lipid Composition in Overweight or Obese Adolescent Students
Authors: YIN RUILI , WANG XIAOJING , ZHAO DONG , YU KE , YANG LONGYAN 
Published: June 2020

The relationship between dyslipidemia and obesity has been widely reported, but the global lipid profiles associated with the development of obesity remain to be clarified, especially in the East Asia teenage population. We recruited 90 adolescent participants, including 30 individuals with normal body mass index (BMI) (BMI: 16-20), 30 obesity students (BMI: 20-26), and 30 overweight students (BMI: 26-37). All participants were performed anthropometric measurement by using InBody 770. Mass spectrometry coupled with liquid chromatography was applied to detect the global lipidome in the fasting plasma. Clinical biochemical indicators were measured by Cobas Elecsys 601. Both qualitative and quantitative analyses revealed a gradual change in plasma lipid features with obesity students exhibiting characteristics close to overweight students, but they differed significantly from students with normal. The levels of triglyceride, 18-Hydroxycortisol, Isohumulinone A, and 11-Dihydro-12-norneoquassin were up-regulated in obesity group, while phosphatidylcholine (PC), phosphatidylethanolamine (PE), LysoPC, LysoPE, and Phosphatidylinositol were significantly down-regulated in obesity group than in control and overweight individuals. Then conducted venn diagram and selected 8 significant metabolites from the 3 paired comparisons. Most of the selected features significantly correlated with anthropometric measurement.
In conclusion, the altered plasma lipidome in Chinese obesity and overweight students suggested that lipid features may provide a basis for evaluating risk and monitoring obesity development.

Disclosure
R. Yin: None. X. Wang: None. D. Zhao: None. K. Yu: None. L. Yang: None.

Funding
National Natural Science Foundation of China (81800768)
2317-PUB: Levels of Vaspine and Omentine in Circulation and Topography of Adipose Tissue Deposition
Authors: KRAVCHUN NONNA , IEFIMENKO TETIANA , MISURA KATHERINE , ALHAMID MOHAMAD 
Published: June 2020

Introduction: Vaspin and Omentin are adipokines that cause peripheral tissue sensitivity to insulin and are a link between type 2 diabetes and obesity. The question of the consequences of excess accumulation in visceral and glutemoral adipose tissue at the level of vaspin and omentin remains unclear today.
Objective: the content of vaspine and omentin in blood depending on the degree and topography of the deposition of adipose tissue in representatives of the Ukrainian population.
Materials and Methods: An enzyme-linked immunosorbent assay of vaspin and omentin levels was determined. Abdominal obesity was determined by the circumference of the waist: in women it was more than 0.80 m, in men - more than 0.94 m.
Results: A direct correlation was found between blood pressure and waist and a hip examination (r = 0.4314, (p &lt;0.001) and r = 0.5130 (p &lt;0.001), respectively). The presence of a relationship between the degree of deposition of adipose tissue on the anterior abdominal wall and the level of circulating omentin confirms the presence of an inverse correlation between this parameter and RT (r = -0.397; p &lt;0.001). The degree of correlation between the level of circulating omentin and the hips was determined: it was higher than with the waist (r = -0.532, p &lt;0.001). The latter indicates that not only visceral, but also gluteal-femoral adipose tissue is displayed at the level of circulating omentin.
Conclusion: The decisive role of abdominal obesity in pathological changes in the production of vaspin and omentin has been confirmed. At the same time, it was found that the circumference of the hips is an informative criterion for changes in the circulation of omentin and vaspin.

Disclosure
N. Kravchun: None. T. Iefimenko: None. K. Misura: None. M. Alhamid: None.
232-LB: Chronic Adrenergic Stimulation of Human Brown and White Adipose Tissue Increases Bile Acid Synthesis Enzyme Expression and Plasma Bile Acid Levels
Authors: ZHU KENNETH Y. , LEA HANNAH J. , CERO CHERYL , CYPESS AARON M. 
Published: June 2020

Beyond enabling digestion of fats, bile acids (BA) are now known to be receptor ligands that improve glucose and lipoprotein metabolism. Classically, regulation of BA synthesis has thought to be exclusively hepatic, but emerging evidence in rodents indicates that BA production is influenced by sympathetic activation of brown and white adipose tissue (BAT and WAT). We recently showed that chronic activation of BAT and WAT in adult women with the β3-adrenergic receptor (AR) agonist mirabegron increased HDL cholesterol, insulin sensitivity, and total plasma BA. The largest increases were with the primary unconjugated cholic and chenodeoxycholic acids. We hypothesized that chronic adrenergic stimulation increased BA synthesis enzymes in adipose tissue, thus increasing flux through these pathways. We therefore measured expression levels of BA synthesis enzymes from patients undergoing elective surgery in the following tissues: subcutaneous WAT (scWAT), and from pheochromocytoma (pheo) patients who have chronic adrenergic activation, scWAT (PhW) and peri-renal BAT (PhB). These were compared to cadaveric liver. Liver had higher expression levels of BAAT, which conjugates BA. However, expression levels among liver, scWAT, PhW, and PhB were similar for the enzymes initiating BA synthesis (CYP7A1) and other intermediate enzymes (CYP7B1, AKR1D1, AMCAR). Furthermore, pheo patients had higher levels of CYP27A1, which initiates the acidic/alternative BA synthesis pathway. These data suggest that regulation of plasma BA is not exclusive to the liver. Rather, human BAT and WAT likely contribute to the BA pool, and chronic adrenergic activation may shift the equilibrium toward primary unconjugated species to improve glucose and lipoprotein metabolism.

Disclosure
K.Y. Zhu: None. H.J. Lea: None. C. Cero: None. A.M. Cypess: None.
242-OR: Rare Genetic Variants in the Islet Regulome Contribute to the Heritability of Type 2 Diabetes Risk
Authors: MANNING ALISA ,   TOPMED DIABETES WORKING GROUP
Published: June 2020

Genome-wide association studies of type 2 diabetes (T2D) show common variant heritability is enriched within gene regulatory regions of pancreatic islets. Whole genome sequence (WGS) data in cohorts allows us to determine the contribution of the complete spectrum of genetic variation to the heritability of T2D risk, including rare alleles that have recently arisen in the human population. A variant-based heritability analysis can partition disease risk into environmental and genetic components, and the genetic components can be further decomposed by variant frequency (a proxy of variant age) and pancreatic islets regulatory annotation (a proxy of functional relevance to T2D risk). We hypothesized that rare, non-protein coding variants would have a significant contribution to the heritability of T2D risk and that further restricting to gene regulatory regions active in pancreatic islets would also produce significant heritability estimates. We conducted a WGS heritability analysis (GREML method in GCTA software) of T2D risk in 15109 unrelated individuals (2215 with T2D; 12894 controls) of European ancestry from the NHLBI’s TOPMed study. We partitioned variants with allele count &gt; 5 by minor allele frequency (MAF): rare variants [MAF&lt;0.1%], and annotation: all non-coding variants, and non-coding variants in pancreatic islets regulatory regions. We further partitioned variants by linkage disequilibrium properties to reduce estimate bias. One of the rare, "all non-coding" variants sets showed a relatively large liability-scale heritability estimate (h2=0.6, P&lt; 0.001). Further significant estimates in rare variants were from active transcription start sites (h2=0.1, P&lt;0.01); active promoters (three variant sets with h2=0.07, 0.08 and 0.1, P&lt;0.05, after body mass index adjustment) and active enhancers from promotor capture Hi-C models (h2=0.07, P&lt;0.05). These findings show a role for rare, non-coding variants in the genetic architecture of type 2 diabetes.

Disclosure
A. Manning: None.

Funding
National Institutes of Health (K01DK107836)
249-OR: A Randomized, Double-Blind Phase 2 Trial of the CXCR1/2 Inhibitor Ladarixin in Adult Patients with New-Onset Type 1 Diabetes
Authors: BOSI EMANUELE , POZZILLI PAOLO , GIORGINO FRANCESCO , COSSU EFISIO , LINN THOMAS , ROSE LUDGER , KEYMEULEN BART , GILLARD PIETER , DAFFONCHIO LUISA , RUFFINI PIER A. , ALLEGRETTI MARCELLO , PIEMONTI LORENZO 
Published: June 2020

In preclinical studies, the investigational CXCR1/2 inhibitor ladarixin was shown to prevent and reverse diabetes in NOD mice. On this basis, a multicenter clinical trial in adult patients with new onset type 1 diabetes (T1D) was conducted (NCT02814838).
Patients (Table 1) were randomized 2:1 to ladarixin (LDX) 400 mg BID or placebo (PLA) and randomized within 100 days from first insulin administration.
Patients received 3 treatment cycles, each consisting of 14 days on and 14 days off drug, and were then followed up for 12 months. The difference between LDX and PLA in mixed meal tolerance test (MMTT) AUC of C peptide was statistically significant neither at week 13 (primary endpoint, p=0.330) nor at week 26 (p=0.404). Conversely, at week 26 patients having HbA1c&lt;7% and daily insulin requirements &lt;0.50 IU/kg were 76.6% and 45.8% in LDX and PLA, respectively (p=0.0095).
In patients with baseline fasting C-peptide below the median value of trial population (prespecified subgroup analysis), the difference between LDX and PLA in MMTT AUC of C peptide trended at week 13 (p=0.111) and reached statistical significance at week 26 (p=0.041).
Adverse Reactions occurring in ≥10% of patients included dyspepsia (16% LDX vs. 0% PLA) and headache (16% LDX vs. 15.4% PLA).
Although primary endpoint was not met, secondary and subgroup analyses suggest further investigation of LDX in new onset T1D may be warranted.

Disclosure
E. Bosi: Consultant; Self; Abbott, Eli Lilly and Company, Medtronic, Novartis AG, Roche Diabetes Care, Sanofi. P. Pozzilli: Advisory Panel; Self; Abbott, AstraZeneca, Eli Lilly and Company. Research Support; Self; Medtronic, Sanofi. F. Giorgino: Advisory Panel; Self; AstraZeneca, Eli Lilly and Company, Novo Nordisk Inc., Roche Diabetes Care, Sanofi. Consultant; Self; AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., LifeScan, Inc., MedImmune, Merck Sharp & Dohme Corp., Roche Diabetes Care, Sanofi. Research Support; Self; Eli Lilly and Company, LifeScan, Inc., Takeda Development Centre Europe Ltd. E. Cossu: None. T. Linn: Advisory Panel; Self; Dompe Pharmaceuticals. L. Rose: None. B. Keymeulen: None. P. Gillard: None. L. Daffonchio: Employee; Self; Domp farmaceutici S.p.A. P.A. Ruffini: Employee; Self; Domp farmaceutici S.p.A., Domp farmaceutici S.p.A. M. Allegretti: Employee; Self; Domp Farmaceutici spa. L. Piemonti: Advisory Panel; Self; AstraZeneca, Roche Diabetes Care, Semma Therapeutics, Inc.
250-OR: Hypoproliferative CD8 Effector Memory (EM) Subsets Are Linked to Preservation of C-Peptide in Alefacept-Treated Recent-Onset Type 1 Diabetes (T1D) Subjects
Authors: SERTI ELISAVET , DIGGINS KIRSTEN E. , MUIR VIRGINIA S. , LU TINGTING , BALMAS ELISA , NEPOM GERALD T. , LONG S. ALICE , LINSLEY PETER 
Published: June 2020

In the ITN T1DAL trial, alefacept (LFA3-Ig fusion protein) preserved endogenous insulin production in 30% of recent onset T1D patients for two years after therapy. Although patients experienced immune alterations following treatment, including depletion of CD2hi effector T-cells and preservation of CD2lo regulatory T-cells, these effects were not predictive of overall response or outcome. However, using an unbiased approach and a combination of flow cytometry, RNA-sequencing, and CyTOF, we were able to identify a CD8 T-cell signature of response at ∼two years following treatment. Preservation of C-peptide was associated with a large transcriptional module (n= 738 genes; R= 0.4, p= 0.023) including activation and exhaustion-associated genes identified at two years following treatment using WGCNA. This module was further dissected into two distinct CD8 EM populations through correlation with hierarchically gated and clustered cytometry data. While both populations expressed multiple inhibitory receptors (IR) (TIGIT, KLRG1, T-bet, EOMES) and were hypo-responsive in in vitro proliferation assays, they were distinguished by higher expression either of markers seen in cytotoxic and terminally differentiated effector T cells (CD57+ Granzyme B+) or of PD-1. Gene expression signatures of both the CD57 and PD-1 subsets significantly overlapped with a previously identified partial exhaustion signature (p= 0.005, for both subsets). Our findings demonstrate an association of partially exhausted memory CD8 T-cells with beneficial clinical response to alefacept as we observed previously with teplizumab (anti-CD3) treatment. Our findings also demonstrate linkage of two phenotypically distinct IR-high, hypo-proliferative CD8 EM T-cell subsets with beneficial response. Together, our findings suggest that pathways regulating proliferation of EM CD8 cells may lead to successful immune interventions for T1D.

Disclosure
E. Serti: None. K.E. Diggins: None. V.S. Muir: None. T. Lu: None. E. Balmas: None. G.T. Nepom: None. S. Long: None. P. Linsley: Consultant; Self; Bristol-Myers Squibb.

Funding
National Institute of Diabetes and Digestive and Kidney Diseases/National Institute of Allergy and Infectious Diseases (UM1AI109565)
252-LB: Insulin Secretion Pathway Genes Correlate with Markers Associated with Viral Infections in Human Islets
Authors: NELSON GRACE , LENCHIK NATALIYA I. , RODRIGUEZ-CALVO TERESA , BALCACEAN DIANA , GERLING IVAN C. ,   NPOD-V
Published: June 2020

Type 1 diabetes (T1D) is an autoimmune disease hypothesized, to be enhanced or triggered by viral infections. In this study we used organ donor pancreas to isolate islets that expressed known signs of viral infections, and subsequently investigated pathways that may be secondarily affected through analysis of gene expression profiles. Using samples provided by the Network for Pancreatic Organ Donors with Diabetes, (nPOD), islets were collected, by laser-capture, from nondiabetic donors (Control), autoantibody positive nondiabetic donors (AB+) and donors with T1D (T1D). Islets were categorized based on the presence of markers associated with viral infection (VIMs), HLA, Mx1, dsRNA, and PKR, identified via immunohistochemical staining. From each donor islets were pooled based on the number of VIMs (0 VIMs, 1 VIMs or ≥2VIMs). RNA was extracted, and microarray used to assess transcriptomes. GeneSpring software was used to generate a list of genes with differential expression between donors/VIMs. Using WebGestalt we identified pathways enriched by this list. A total of 383 genes with a fold change of ≥1.1 and p-value=0.001 had differential expression between islets with 0 VIMs and ≥1 VIMs. Pathway analysis showed strongest enrichment for genes in the insulin secretion pathway (p&lt; 5.7E-5). As an example; CACNA1C and ABCC8 (important for the secretion of insulin), had lower expression in the islets with ≥1VIMs compared to 0 VIMs (p&lt;0.03). CACNA1C also had lower expression in islets with 0 VIM vs. ≥1VIMs when only comparing samples from control donors (p=0.02). Islets selected for high expression for genes associated with viral infection had decreased expression of genes important for insulin secretion. This may suggest a pattern of dedifferentiation and/or functional impairment of beta cells in the setting of viral infection.

Disclosure
G. Nelson: None. N.I. Lenchik: None. T. Rodriguez-Calvo: None. D. Balcacean: None. I.C. Gerling: None.

Funding
JDRF; National Institute of Diabetes and Digestive and Kidney Diseases; Lebonheur Children’s Hospital