bluuebunny/crossref_metadata_2025_split huggingface dataset:
1676-P: Immune Responses to Insulin B:9-23–Related Peptides Were Observed in Fulminant Type 1 Diabetes
Authors: OIKAWA YOICHI , SATOMURA ATSUSHI , HAISA AKIFUMI , SUZUKI SEIYA , SHIMADA AKIRA 
Published: June 2020

Most type 1 diabetes (T1D) is considered to be a T helper (Th)-1 type anti-islet autoimmune disease. Recently, we reported that the immune responses to insulin B9-23-related peptides were strengthened in T1D. Meanwhile, fulminant T1D (FT1D), a novel subtype of T1D characterized by extremely rapid onset and complete deficiency of insulin due to destruction of pancreatic β-cells, has been believed to be caused by viral infection as the etiological mechanism; but, whether the etiology associates with anti-islet immune reactions or not remains unknown. This study aims to investigate whether the immune responses to insulin B9-23-related peptides associate with FT1D. Peripheral mononuclear cells (MNCs) were obtained from 72 patients with acute-onset T1D (AT1D; age, 46.2 ± 14.7 y/o; disease duration (DD), 8.8 ± 10.7 years), 15 patients with FT1D (age, 46.1 ± 14.2 y/o; DD, 4.7 ± 8.2 years) and 25 patients with type 2 diabetes (T2D; age, 63.3 ± 13.1 y/o; DD, 17.0 ± 10.2 years). The MNCs were stimulated by insulin B9-23, B10-24, B11-25 and B12-26 for 24 hours. Subsequently, ELISpot assay was performed for investigating a frequency of interferon (IFN)-gamma (g)-producing MNCs. The maximum number of IFN-g spots in the peptides-related data of each subject was used for analysis and expressed by the number of spot-forming cells (SFCs) per 2.5×105 cells. As a result, the spot number was significantly higher in AT1D (2.6 ± 3.0 SFCs) and FT1D (2.5 ± 2.6 SFCs) as compared to T2D groups (1.0 ± 1.1 SFCs) (P &lt;0.05 by t-test), suggesting that Th1-type immune responses to insulin B9-23-related peptides were observed in FT1D, and that the degree of response was comparable in the two groups, suggesting the association between anti-islet autoimmunity and FT1D. This cross-sectional study could not reveal an association between the Th1-type immune responses and disease activity which may be inferred from the residual serum C-peptide levels or disease duration, which warrant further investigation by accumulating more subjects.

Disclosure
Y. Oikawa: None. A. Satomura: None. A. Haisa: None. S. Suzuki: None. A. Shimada: Advisory Panel; Self; Astellas Pharma Inc. Speaker’s Bureau; Self; Eli Lilly Japan K.K., Novo Nordisk Inc., Sanofi-Aventis.
168-OR: The Inverse Association between Cardiorespiratory Fitness and Inflammation Is Modified by Plasma Levels of Omega-3 Polyunsaturated Fatty Acids
Authors: FARLEY GABRIELLE , RIGGS DANIEL W. , BHATNAGAR ARUNI , HELLMANN JASON 
Published: June 2020

Moderate intensity exercise has been shown to prevent the development and progression of chronic inflammatory diseases, including type 2 diabetes. Likewise, physical fitness is associated with reduced inflammation, morbidity, and mortality. Nonetheless, factors that contribute to the anti-inflammatory effect of physical fitness remain largely undefined. Using data collected from 437 participants in the 2003-2004 National Health and Nutrition Examination Survey, we assessed how the omega-6/omega-3 fatty acid ratio (ω-6/ω-3) modifies the relationship between cardiorespiratory fitness (VO2max) and inflammation (C-reactive protein; CRP). Participants were 45% female with an average age of 32.9 years and with a BMI of 27.7 kg/m2. In adjusted models for CRP, we observed significant interactions between VO2max and ω-6/ω-3 quartiles. Participants with the highest quartile of ω-6/ω-3 showed a positive association between VO2max and CRP levels. Conversely, the lowest quartiles of ω-6/ω-3 were negatively associated with VO2max and CRP, ranging from -22.4% to -33.7% CRP per 10 ml/kg/min VO2max compared with the highest ω-6/ω-3 quartile (interaction p-value= 0.0149). This negative relationship was strengthened when alternatively modeling the interaction between total ω-3 levels and VO2max (interaction p-value=0.0002), wherein higher ω-3 levels resulted in a stronger negative association between VO2max and CRP. This was predominantly driven by alpha-linoleic acid (ALA), docosahexaenoic acid (DHA), and eicosapentaenoic acid (EPA), but not docosapentaenoic acid (DPAn-3) levels. We also observed negative associations between circulating levels of monocytes and VO2max for participants with lower ω-6/ω-3 or higher ω-3 EPA levels. Collectively, these data suggest that the anti-inflammatory effect of physical fitness is enhanced with higher plasma levels of ω-3 fatty acids.

Disclosure
G. Farley: None. D.W. Riggs: None. A. Bhatnagar: None. J. Hellmann: None.

Funding
National Institutes of Health (GM127495, GM127607)
1680-P: A Novel Immunomodulatory Combination That Decreases Adipose Tissue and Systemic Inflammation and Improves Metabolism in Dio Mice
Authors: SATER ZAHRAA A. , CERO CHERYL , LEA HANNAH J. , ZHU KENNETH Y. , GAVRILOVA OKSANA , CYPESS AARON M. 
Published: June 2020

β3-adrenoceptors (B3AR) are major activators of lipolysis and thermogenesis in adipose tissue (AT). Obesity increases oxidative stress and AT inflammation leading to the attenuation of β3AR signaling. Alpha-lipoic acid (ALA) is a potent antioxidant. We hypothesized that the combination of ALA with CL316,243 (CL) -a selective B3AR agonist- restores the immune balance in diet-induced obesity (DIO) leading to increased B3AR function. We separated 40 DIO C57BL/6 male mice into 4 groups (n=10; individually caged) and treated them for five weeks with 1. vehicle control [IP (saline) + PO (drinking water)] 2. CL alone (0.01 mg/kg IP daily) 3. ALA alone (250 mg/kg orally in drinking water) 4. ALA+CL combination. All mice received ad-lib HFD (60% kcal fat). Systemic and epididymal white AT (epi-WAT) inflammation was decreased in ALA+CL compared to other groups, with lower serum IL-6 and IL-17 and lesser crown-like structures (CLS) on IHC for F4/80 and CD11c (M1-MΦ). Both IHC staining and mRNA expression of CD206 (M2-MΦ marker) confirm increased M2 MΦ’s in ALA+CL compared to other groups. Food intake was similar in all groups, yet mice receiving ALA+CL had a better metabolic profile: Lower body weight [+1.7 vs. -2.5g (-7%); p&lt;0.01] and % body fat (+1.0 vs. -2.8g; (-9%), p&lt;0.001) compared to controls preserved lean mass. Moreover, ALA+CL showed improved glucose tolerance compared to controls (intraperitoneal glucose tolerance test; AUC glycemia 31,033 ± 950 vs. 25,919 ± 620 min*ml/dl; p&lt;0.01; AUC insulinemia 441.2 ± 55.7 vs. 279.3 ± 42.2 min*ng/ml; P&lt;0.001). The downstream function of B3AR(lipolysis and thermogenesis) in WAT was improved in the combination group compared to all groups evidenced by markedly higher activated lipases and UCP-1 on Western blots (WB), and ucp-1 expression on mRNA analysis. Our study suggests that ALA, in combination with CL, modulates the immune response to obesity and augment catecholamine effect on lipolysis and thermogenesis in WAT.

Disclosure
Z.A. Sater: None. C. Cero: None. H.J. Lea: None. K.Y. Zhu: None. O. Gavrilova: None. A.M. Cypess: None.
1681-P: Alpha-1 Antitrypsin Suppresses Macrophage Activation Post Islet Transplantation
Authors: GOU WENYU , STRANGE CHARLIE , WANG HONGJUN 
Published: June 2020

Macrophage activation contributes to islet graft loss after transplantation by secretion of proinflammatory cytokines. We showed in previous studies that treatment with alpha-1 antitrypsin (AAT) improved human islet survival after transplantation into the livers of streptozotocin-induced diabetic NOD-SCID mice. To further understand the cellular targets of AAT, we isolated macrophages from control or AAT-treated recipients at day 1 post transplantation (PT) and compared their mRNA profile using RNAseq analysis. 344 genes were changed in AAT-treated recipients compared to controls. Clusters of the inflammatory response, chemokine activity and cytokine and cytokine receptor Gene Otology categories and pathways were significantly affected by AAT-treatment. In vitro, AAT suppressed IFN-γ-induced Raw267.4 cell activation via suppression of STAT1 phosphorylation and iNOS production. This was translated in vitro when human islets were co-transplanted with IFN-γ-treated Raw cells, in the presence or absence of AAT, into diabetic NOD-SCID mice in which macrophages were depleted by liposome. At day 21, all macrophage-depleted diabetic recipients receiving islets alone achieved normoglycemia (n=6). In contrast, only 55.6% (n=9, p=0.001) receiving islets and IFN-γ-treated Raw cells achieved normoglycemia, compared to 90% receiving AAT-treated IFN-γ Raw cells (n=10, p=0.003), suggesting AAT improved islet graft survival via suppressing macrophage activation. The effect of AAT was further confirmed in a major mismatch allogeneic islet transplantation model (Balb/c, H-2b to C57BL/6 H-2d mice) in which macrophage activation contributed to primary nonfunction (PNF) of the transplanted islets. In this model, PNF occurred in 16.7% of AAT-treated recipients (n=6) but in 40% of controls (n=5) when 400 islets were transplanted.
In summary, this study further identified that AAT contributed to islet graft survival after transplantation by suppressing macrophage activation.

Disclosure
W. Gou: None. C. Strange: None. H. Wang: None.

Funding
National Institutes of Health (DK105183, DK120394, DK118529)
1685-P: Islet Encapsulation with Tannic Acid Delays Allogeneic Islet Graft Rejection
Authors: BARRA JESSIE , KOZLOVSKAYA VERONIKA , KHARLAMPIEVA EUGENIA , TSE HUBERT M. 
Published: June 2020

Type 1 diabetes (T1D) is defined by the destruction of insulin-producing cells. Islet transplantation can restore glucose homeostasis and prevent serious hypoglycemic events; however, long-term graft survival is low. Oxidative stress can mediate islet transplantation rejection and suppressing oxidative stress with nanothin islet encapsulation materials comprised of alternating layers of tannic acid (TA), an immunomodulatory antioxidant, and poly(N-vinylpyrrolidone) (PVPON), a scaffold polymer, may allow for improved graft survival without systemic immunosuppression. We hypothesize that (PVPON/TA)-encapsulation of islets will prolong graft function following allotransplantation into immunocompetent recipients without utilizing systemic immunosuppression. Allogeneic transplantation of encapsulated C57BL/6 islets into diabetic non-obese diabetic (NOD) mice (n=12) significantly delayed graft failure by more than 90 days compared to controls and significantly reduced free radical-adduct formation produced by neutrophils and macrophages at the graft site. Gene expression displayed a 5- and 20-fold increase of anti-inflammatory genes Retnla and Arg1, respectively, and 3-fold reductions in Ccl5, Cxcl10, Prf1, and Grzmb mRNA accumulation in (PVPON/TA) versus control grafts. Flow cytometry analysis of encapsulated grafts contained fewer numbers of infiltrating dendritic cells and CD8 T cells and reduced frequencies of CCL5+, CXCR3+, MHC-II+ macrophages and perforin+ CD8 T cells by 3-fold compared to controls. Encapsulated grafts also reduced the frequencies of TNF+ and MHC-II+ dendritic cells by 2-fold. Additionally, encapsulation increased the frequency of arginase-1+ and CD206+ alternatively-activated macrophages 4-fold, suggesting that (PVPON/TA)-encapsulation may be a promising technique to prolong clinical islet graft function by skewing macrophage phenotypes and suppressing cytotoxic T cell effector responses in the absence of systemic immunosuppression.

Disclosure
J. Barra: None. V. Kozlovskaya: None. E. Kharlampieva: None. H.M. Tse: None.

Funding
National Institute of Diabetes and Digestive and Kidney Diseases (R01DK099550); JDRF (SRA-2016-270-S-B, JDRF 2-SRA-2019-692-S-B); National Science Foundation (NSF-DMR 1608728); National Institute of General Medical Sciences (T32GM008111)
1688-P: Peak Insulin Drip Rate Associated with Decreased Infections Post–Heart Transplant
Authors: UPADRASTA GAUTHAM , KUMAR SMITA J. , WALLIA AMISHA , MOLITCH MARK 
Published: June 2020

Hyperglycemia is associated with infection and rejection in post-solid organ transplant (Tx) patients (pts), but little is known about peak insulin requirements and morbidity. We previously reported an unexpected finding in a retrospective cohort study of 207 liver Tx pts that evaluated the relationship to peak IV insulin requirements and subsequent morbidity. Pts were separated into quartiles (Q) by peak insulin drip rate [U/hr]. In that study, pts in Q4 of insulin requirements had significantly reduced infections out to 6 months post-Tx compared to Q1-3.
In the present study, we sought to replicate these findings in 188 heart Tx pts. Peak drip rate Qs are shown in the Figure below. Compared to Q1-3, Q4 pts were similar in age (53 ± 10.7 vs. 50.3 ± 9.4) and BMI (30.4 ± 6.4 vs. 29.2 ± 6.8), but more had DM (42.6% vs. 38.3%, p=0.003). Q4 pts had significantly fewer infections (19.1% vs. 53.9%, p&lt;0.000001) up to 6 mos but similar rejections (4.3% vs. 11.3%, p=0.15). Also, we showed that avg blood glucose levels were not significantly different between Q1-3 and Q4 (p = 1.03).
These data in heart Tx pts confirm our findings in liver Tx pts that pts requiring higher insulin doses for glycemic control have decreased infections out to 6 months post Tx. We hypothesize that pts able to mount a greater counterregulatory hormone response to surgery may be healthier and able to better respond to infection. Alternatively, higher insulin doses could be protective.

Disclosure
G. Upadrasta: None. S.J. Kumar: None. A. Wallia: Research Support; Self; Eli Lilly and Company, Novo Nordisk Inc., UnitedHealth Group. M. Molitch: Advisory Panel; Self; Janssen Pharmaceuticals, Inc., Merck & Co., Inc., Pfizer Inc. Consultant; Self; AstraZeneca. Research Support; Self; Novartis Pharmaceuticals Corporation, Novo Nordisk Inc.
1690-P: In Vitro Differentiation of Pancreas-Derived Mesenchymal Stem Cells into Insulin-Producing Beta Cells
Authors: KALIVARATHAN JAGAN , SARAVANAN PRATHAB BALAJI , LEVY MARLON F. , KANAK MAZHAR A. 
Published: June 2020

Differentiation of insulin-producing cells (IPCs) from stem cells has been widely studied as an alternative source for cell transplantation therapy in diabetes. Mesenchymal stem cells (MSCs) are a useful therapeutic option for islet transplants due to their immunomodulatory properties and differentiation potential. In this study, we introduce a method for in vitro differentiation of pancreatic MSCs into IPCs. Research-grade pancreas from cadaveric donors was used to isolate islets, and the COBE remnant after islet purification was used for the generation of pancreatic MSCs. The remnant tissue was cultured in RPMI medium and plastic to isolate tissue-resident MSCs. Surface marker expression analysis and differentiation assays were performed to characteristically confirm that the cells obtained were in fact MSCs. Osteogenic, adipogenic and chondrogenic differentiation of MSCs were induced by StemPro® differentiation kits. The protocol for the differentiation of IPCs from stem cells was adopted from Pagliuca et al. 2014. Similarly, we have treated the pancreatic MSCs with molecules and growth factors at different stages of development to produce IPCs. The insulin secretion from differentiated IPCs also analyzed at all stages. The culture of remnant tissue after islet isolation resulted in plastic-adherent cells of fibroblast-like morphology after 3 passages. Flow cytometry analysis showed homogeneous expression of markers (CD90, CD73, CD29, CD105) and negative expression of CD14, CD34, and CD45 as classically characterized by cultured MSCs. Interestingly, there was a significant increase in insulin levels up to 5 and 100 folds in the media at developmental stages S5 (P= 0.0686) and S6 (P &lt;0.0001) respectively. Our results reveal that MSCs can be directly differentiated into IPCs. Human pancreas-derived MSCs may serve as an alternative source of insulin-producing beta cells that can be transplanted to type 1 diabetes patients.

Disclosure
J. Kalivarathan: None. P. Saravanan: None. M.F. Levy: None. M.A. Kanak: None.
1692-P: Ablation of Endothelial ROCK2 Promotes Fat Browning and Improves Metabolic Dysfunction
Authors: TAKEDA YUSUKE , MATOBA KEIICHIRO , KAWANAMI DAIJI , NAGAI YOSUKE , KANAZAWA YASUSHI , NISHIMURA RIMEI 
Published: June 2020

Intraperitoneal fat is considered a particularly important risk factor for impaired glucose tolerance, dyslipidemia, and coronary heart diseases. However, the underlying mechanisms in obesity are not fully understood. The small GTPase Rho and its downstream effector, Rho-kinase (ROCK), regulate various cellular functions, including organization of the actin cytoskeleton, cell adhesion and expression of genes. Recent studies have shown that Rho/ROCK signaling is strongly related to diabetic vascular complications. ROCK has two isoforms, ROCK1 and ROCK2. Systemic gene deletion studies in mice suggest that these isoforms have distinctive roles in regulating cellular function. In this study, we investigated specific roles of endothelial ROCK2 in the progression of obesity. We mated ROCK2 floxed mice with VE-cadherin-cre mice to generate endothelial-ROCK2 knockout mice (ER2KO). ER2KO mice are resistant to weight gain and glucose intolerance induced by high fat diet. White adipose tissue (WAT) weight was lower in ER2KO mice compared with wild type mice. Histological analysis revealed that adipose droplets were smaller in ER2KO than wild type mice. Browning, the conversion of WAT to a beige phenotype, activates thermogenic function, suppresses obesity and improves glucose and lipid metabolism. Interestingly, we observed an increase of mRNA expression of browning marker including PPARα, CIDEA, PRDM16, UCP1, and specific markers of M2 macrophages in WAT of ER2KO mice regardless of whether they had been fed a normal chow or high fat diet. Collectively, the present study suggests that endothelial ROCK2 regulates glucose and lipid metabolism by suppressing browning of WAT. ROCK2 could be an important therapeutic target against obesity.

Disclosure
Y. Takeda: None. K. Matoba: Research Support; Self; Sanofi K.K., Takeda Pharmaceutical Company Limited. D. Kawanami: None. Y. Nagai: None. Y. Kanazawa: None. R. Nishimura: Research Support; Self; Boehringer Ingelheim Pharmaceuticals, Inc. Speaker’s Bureau; Self; Abbott, AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Eli Lilly Japan K.K., Medtronic, Merck & Co., Inc., Novo Nordisk Inc., Sanofi K.K., Takeda Pharmaceutical Company Limited.
1694-P: Beta-Arrestin 1 Suppresses Myogenic Reprogramming of Brown Fat to Maintain Euglycemia
Authors: PYDI SAI PRASAD , JAIN SHANU , BARELLA LUIZ F. , MEISTER JAROSLAWNA , WANG LEI , CUI YINGHONG , GAVRILOVA OKSANA , WESS JÜRGEN 
Published: June 2020

Background: Adipocyte dysfunction links obesity to type 2 diabetes (T2D). To develop novel antidiabetic drugs, it is essential to delineate the signaling pathways that regulate adipocyte function under physiological and pathophysiological conditions. Recent studies have shown that β-arrestin-1 and -2 (barr1 and barr2, respectively), two intracellular signaling proteins known for their ability to modulate signaling via G protein-coupled receptors (GPCRs), regulate many important metabolic functions, sometimes even in a GPCR-independent fashion.
Objective: To understand the role of barr1 in adipocyte function, obesity, and whole body glucose and energy homeostasis.
Methods: To assess the role of barr1 in regulating adipocyte function, we used Cre-lox technology to generate mutant mice that lacked barr1 selectively in adipocytes(adipo-barr1-KO). These mutant mice, together with their control littermates, were subjected to a series of metabolic tests. Moreover, we used various molecular and biochemical techniques to probe the molecular mechanism through which barr1 modulates adipocyte function.
Results: We found that adipo-barr1-KO mice showed greatly impaired glucose tolerance and insulin sensitivity when consuming an obesogenic diet. In contrast, transgenic mice that overexpressed barr1 in adipocytes were protected against the metabolic deficits caused by a high-calorie diet. At the cellular level, barr1 deficiency led to a myogenic re-programming of brown adipose tissue (BAT), causing elevated plasma myostatin levels, which in turn led to impaired insulin signaling in peripheral tissues. Additional in vivo studies indicated that barr1-mediated suppression of myostatin expression by BAT is required for maintaining euglycemia.
Conclusion: Our data suggest that strategies aimed at enhancing barr1 expression or activity in BAT may prove beneficial for the treatment of T2D.

Disclosure
S. Pydi: None. S. Jain: None. L.F. Barella: None. J. Meister: None. L. Wang: None. Y. Cui: None. O. Gavrilova: None. J. Wess: None.

Funding
National Institute of Diabetes and Digestive and Kidney Diseases
1698-P: Obesity-Induced Increase in Cystatin C Alleviates Tissue Inflammation and Hepatic Insulin Resistance
Authors: DEDUAL MARA A. , WUEEST STEPHAN , CHALLA TENAEGENE DELESSA , LUCCHINI FABRIZIO , AEPPLI TIM , BORSIGOVA MARCELA , MAURACHER ANDREA A. , VAVASSORI STEFANO , SCHMID JANA PACHLOPNIK , BLÜHER MATTHIAS , KONRAD DANIEL 
Published: June 2020

Removal of one kidney (uninephrectomy; UniNx) in mice reduced high fat-diet (HFD)-induced adipose tissue inflammation thereby improving adipose tissue and hepatic insulin sensitivity. Such finding was accompanied by increased circulating cystatin C levels in UniNx compared to sham-operated mice. Importantly, cystatin C may have anti-inflammatory properties, and circulating cystatin C levels were reported to positively correlate with obesity as well as type 2 diabetes. However, the causal relationship of such correlation remains unclear. Herein, HFD feeding of cystatin C-deficient (CysC KO) mice aggravated obesity-associated adipose tissue dysfunction and inflammation, as reflected by an accumulation of pro-inflammatory monocytes and elevated expression of pro-inflammatory mediators. Moreover, markers of adipocyte differentiation were decreased. Similarly to findings in adipose tissue, expression of pro-inflammatory cytokines was increased in liver and skeletal muscle of HFD-fed CysC KO mice. In line, HFD-induced hepatic insulin resistance and impairment of glucose tolerance were further aggravated in knockout mice. Consistently, chow-fed CysC KO mice were more susceptible to lipopolysaccharide (LPS)-induced adipose tissue inflammation. In people with obesity, circulating cystatin C levels correlated negatively with mRNA expression of pro-inflammatory cytokines such as Hif1α as well as IL-6. Moreover, healthy (i.e., insulin-sensitive) subjects with obesity depicted significantly higher mRNA expression of cystatin C in subcutaneous white adipose tissue.
In conclusion, cystatin C is upregulated under obesity conditions and thereby counteracts inflammation of peripheral insulin-sensitive tissues and, thus, obesity-associated deterioration of glucose metabolism.

Disclosure
M.A. Dedual: None. S. Wueest: None. T. Challa: None. F. Lucchini: None. T. Aeppli: None. M. Borsigova: None. A.A. Mauracher: None. S. Vavassori: None. J. Pachlopnik Schmid: Advisory Panel; Self; Novartis AG, Swedish Orphan Biovitrum. M. Blüher: Advisory Panel; Self; AstraZeneca, Boehringer Ingelheim Pharmaceuticals, Inc., Lilly Diabetes, Novo Nordisk A/S, Sanofi. Speaker’s Bureau; Self; Amgen, Novartis AG. D. Konrad: Research Support; Self; Novo Nordisk A/S.

Funding
Children’s Research Centre; University Children’s Hospital Zurich; University of Zurich (FK-18-027); Swiss National Science Foundation (310030-160129, 10030-179344); Wolfermann-Nägeli Foundation