bluuebunny/crossref_metadata_2025 huggingface dataset:
1953-P: PPARγ Agonism and FGF-21 Synergize to Robustly Induce Browning of White Fat
Authors: KROON TOBIAS , HARMS MATTHEW J. , NILSSON DANIEL , LINDBLOM ANNA , GENNEMARK PETER , O'MAHONY GAVIN , OSINSKI VICTORIA , MCNAMARA COLEEN A. , BOUCHER JEREMIE
Published: June 2019

Increasing brown adipose tissue abundance or activity or converting white into brown adipocytes holds promise for the treatment of type 2 diabetes and obesity. Several PPARγ agonists, including rosiglitazone, have been shown to robustly induce browning of white adipose tissue (WAT) in vitro but their effects in vivo are less pronounced, while PPARα agonists show modest browning effects both in vitro or in vivo. In the present study, we investigated whether PPARα and PPARγ dual agonism would have synergistic effects in inducing browning of WAT. We found that all tested dual PPARα/γ agonists robustly increased uncoupling protein 1 (Ucp1) expression in both mouse and human preadipocytes, with tesaglitazar giving the largest Ucp1 induction. Notably, tesaglitazar also strongly induced browning of WAT in vivo in both lean and obese mice at thermoneutrality, largely exceeding the increase in Ucp1 observed with rosiglitazone. Mechanistically, we found that selective PPARγ agonism was sufficient for the conversion of white into brown-like adipocytes in vitro, but dual PPARα/γ agonism was superior to selective PPARγ agonism at inducing white fat browning in vivo, at least in part via a PPARα-mediated increase in fibroblast growth factor 21 (FGF-21). In human preadipocytes in vitro, combined treatment with rosiglitazone and FGF-21 resulted in a robust and synergistic increase in UCP1 mRNA levels, superior to rosiglitazone- or FGF-21-monotreatments. Tesaglitazar-induced browning in obese mice was associated with increased energy expenditure, enhanced insulin sensitivity, reduced liver steatosis, and an overall improved metabolic profile compared to rosiglitazone and vehicle control groups.

In conclusion, we found that dual PPARα/γ agonism robustly promotes browning of white fat in vivo, via synergistic action of FGF-21 and PPARγ agonism. Our findings identify a novel opportunity to develop compounds with robust browning of WAT, via dual modulation of PPARα and PPARγ.

Disclosure

T. Kroon: Employee; Self; AstraZeneca. M.J. Harms: Employee; Self; AstraZeneca. D. Nilsson: None. A. Lindblom: Employee; Self; AstraZeneca. P. Gennemark: Employee; Self; AstraZeneca. G. O'Mahony: Employee; Self; AstraZeneca. V. Osinski: None. C.A. McNamara: None. J. Boucher: Employee; Self; AstraZeneca.
1979-P: GLP-1 Recruits Skeletal Muscle Microvasculature without Impacting Glucose Uptake or Protein Metabolism in Older Men during Postabsorptive Insulin Clamps
Authors: ABDULLA HAITHAM , PHILLIPS BETHAN E. , LIMB MARIE C. , WILKINSON DANIEL J. , JANDOVA TEREZA , CEGIELSKI JESSICA , BASS JOSEPH J. , RANKIN DEBBIE , LEWIS JONATHAN S. , WILLIAMS JOHN , SMITH KEN , ATHERTON PHILIP J. , IDRIS ISKANDAR RAUF
Published: June 2019

Introduction: Ageing muscle is associated with impaired blood flow, “anabolic resistance” to dietary protein, and impaired glucose uptake - culminating in muscle atrophy and insulin resistance. Since GLP-1 has been shown to recruit muscle microvasculature, and due to the presence of its receptors in local tissue(s) (e.g., endothelium, myocytes) we hypothesized that GLP-1 infusions in older people, would positively impact muscle microcirculation, protein and/or glucose metabolism - in an insulin-independent fashion.

Methods: We studied seven older men (∼70±1 y, mean±SEM) in a cross-over design. Baseline measures were taken (first 3 h) prior to insulin and glucose clamps where participants received co-infusions of octreotide, Vamin 14-EF (containing essential amino acids and dextrose) - with or without GLP-1, over the next 3 h. Four muscle biopsies were taken over 90-120 minute intervals. Muscle protein turnover was quantified via A-V balance, and direct incorporation of 13C6 phenylalanine via mass spectrometric techniques. Plasma GLP-1 and insulin were quantified via Milliplex Map, ELISA. Muscle microvascular flow was assessed via contrast enhanced ultrasound. Leg glucose uptake was determined via A-V balance.

Results: GLP-1 increased microvascular flow in +GLP-1 vs. -GLP-1 conditions (increase vs. without GLP-1 infusion: 7.2±4.7 vs. 0.40±0.26-fold, p=0.009) while leg glucose uptake was unchanged. Muscle proteolysis (nb. no effects on synthesis) was equally suppressed vs. basal with GLP-1 (32.1±5.3 vs. 67.4±3.2 nmol.100 g leg-1.min-1, p<0.05) and without GLP-1 (32.77±6.1 vs. 57.9±5.1 nmol.100 g leg-1.min-1 p<0.05, respectively).

Conclusion: GLP-1 recruits muscle microcirculation independent of insulin. A lack of impact of GLP-1 on muscle glucose uptake or protein metabolism, could reflect a requirement for fed-state insulin levels. GLP-1 may hold therapeutic value for muscle microvascular health.

Disclosure

H. Abdulla: None. B.E. Phillips: None. M.C. Limb: None. D.J. Wilkinson: None. T. Jandova: None. J. Cegielski: None. J.J. Bass: None. D. Rankin: None. J.S. Lewis: None. J. Williams: None. K. Smith: None. P.J. Atherton: None. I.R. Idris: None.

Funding

Medical Research Council UK; Arthritis Research UK
1981-P: The Regulation of Incretin Secretion in Baby Rats in Response to a Lipid Meal
Authors: QU JIE , YANG QING , KO CHIH-WEI , LIU MIN , BLACK DENNIS D. , TSO PATRICK
Published: June 2019

Background: Glucose-dependent insulinotropic peptide (GIP) and glucagon-like peptide-1 (GLP-1) are incretin hormones that augment glucose-stimulated insulin secretion, promote pancreatic β-cell proliferation, neogenesis and inhibit β-cell apoptosis. Thus, they are excellent therapeutic targets against type 2 diabetes. Using a lymph fistula rat model, we studied the secretion of incretins into lymph. In adult rats, we found that GIP and GLP-1 concentrations in the fasting lymph were 132 ± 19 pg/ml and 31 ± 8 pM, respectively. After intraduodenal lipid infusion, their concentrations rose to 441 ± 144 pg/ml and 154 ± 23 pM, respectively.

Rationale: However, little is known about nutrient-stimulated secretion of GIP and GLP-1 in baby rats, whose gastrointestinal system and pancreatic β-cells are immature compared to adult rats.

Methods: Using our well-established lymph fistula model with conscious baby rats, just weaned (21 day after birth) and weighing 50-65 g, we assessed the secretion of GIP and GLP-1 in lymph after an intraduodenal challenge of lipid meal.

Results: Fasting GIP and GLP-1 levels were 295.4 ± 48.1 pg/ml and 59.6 ± 16.2 pM, respectively. After lipid infusion, GIP and GLP-1 levels increased to 1052 ± 90.1 pg/ml and 192.0 ± 19.7 pM respectively. Elevated incretin levels in baby rats (relative to adult animals) indicate possible involvement in the maturation of the enteroendocrine system and β-cells. This increase may be also important to ensure the adequate secretion of insulin to regulate blood glucose as the animals shift from a high-fat mother’s milk diet to a chow diet. Further studies are needed to determine the physiological role of the higher incretin levels in baby rats.

Conclusions: Lymph fistula baby rats can serve as an excellent model to study regulation of gut hormones by nutrient absorption at their early developmental stage. This model can also be used to study the influence of mother’s milk on the secretion of gut hormones and their impact on glucose and lipid metabolism in the newborn.

Disclosure

J. Qu: None. Q. Yang: None. C. Ko: None. M. Liu: None. D.D. Black: Advisory Panel; Self; Intercept Pharmaceuticals, Inc. P. Tso: None.

Funding

National Institutes of Health (DK59630)
1986-P: Hippocampal Overexpression of Triggering Receptor Expressed on Myeloid Cell 2 (TREM2) Ameliorates High-Fat Diet-Induced Neurocognitive Impairment via Modulating Microglial M1/M2 Polarization
Authors: WU MIN , XIAO XIAOQIU
Published: June 2019

Objective: Type 2 diabetes mellitus (T2DM) and Alzheimer’s disease (AD) share several common pathophysiological features. It was reported that rare variant of triggering receptor expressed on myeloid cell 2 (TREM2) increases risk of developing AD, suggesting the involvement of TREM2 and microglia in AD development. It is still unknown whether TREM2 is related to neurocognitive impairment of T2DM.

Methods: Male C57BL/6J mice were maintained high fat diet (HFD). TREM2 was overexpressed in hippocampus of mice 40 weeks after HFD feeding, using an adeno-associated virus vector (AAV)-mediated gene delivery. Morris water maze, Y maze-spontaneous alteration test, novel object recognition test, and open field test were performed 32-36 weeks and 48-50 weeks after HFD feeding. Mouse microglial BV-2 cells were used to examine microglial polarization after TREM2 overexpression or knockdown.

Results: HFD feeding caused rapid weight gain, glucose intolerance and significant impairment in learning and memory as well as anxiety-related behaviors. Compared with empty AAV delivery, TREM2 AAV significantly ameliorated glucose homeostasis and neurocognitive impairment without altering body weight in HFD mice. Hippocampal delivery of TREM2 upregulated synaptic protein spinophilin and PSD95, suggesting improvement in synaptic transmission. Microglial M2 polarization markers, such as Arg-1, YM1/2, CD206 and anti-inflammatory cytokines, were positively correlated to TREM2 levels either in the hippocampus of HFD mice or BV-2 cells in vitro.

Conclusion: Hippocampal TREM2 plays an important role in improving obesity-induced cognitive dysfunction and anxiety-like behavior via modulating microglial polarization toward M2 anti-inflammatory subtype. Our study also suggests that TREM2 might be a novel molecular target for the intervention of obesity-associated cognitive decline.

Disclosure

M. Wu: None. X. Xiao: None.

Funding

National Natural Science Foundation of China
1989-P: Ablation of Ghrelin Receptor in Myeloid Cells Attenuates Diet-Induced NAFLD in Mice
Authors: KIM DA MI , LEE JONG HAN , PAN QUAN , NOH JI YEON , WANG HONGYING , HAN HYE WON , WU CHIA-SHAN , GUO SHAODONG , SUN YUXIANG
Published: June 2019

Chronic low-grade inflammation is a hallmark of obesity, which is directly linked to nonalcoholic Fatty Liver Disease (NAFLD) and systemic insulin resistance. Ghrelin receptor, a growth hormone secretagogue receptor (GHS-R), mediates the stimulatory effect of ghrelin on food intake and fat deposition. We previously reported that global ablation of GHS-R in mice decreases adipose inflammation, and GHS-R knockdown in macrophage cell line reduces the expression of pro-inflammatory cytokine genes. To determine the physiological role of GHS-R in macrophages, we generated myeloid-specific GHS-R knockout mice (LysM-Cre;Ghsrf/f). Under diet-induced obesity (DIO), LysM-Cre;Ghsrf/f mice showed reduced circulating inflammatory cytokine levels, attenuated insulin resistance, but no change in body weight. Moreover, DIO-induced NAFLD was ameliorated in LysM-Cre;Ghsrf/f mice, showing reduced inflammation and lipid accumulation in the liver. To understand the regulatory mechanism of macrophage GHS-R in NAFLD pathogenesis and systemic inflammation and insulin sensitivity, we conducted ex vivo studies using bone marrow-derived macrophages (BMDMs). GHS-R deficient BMDMs showed reduced macrophage pro-inflammatory polarization and decreased expression of pro-inflammatory cytokine genes; consistently, exhibiting elevated mitochondrial respiration and improved mitochondrial dynamics. Moreover, the primary hepatocytes treated with conditioned media from BMDMs of LysM-Cre;Ghsrf/f mice showed gene expression of reduced inflammatory cytokines and increased insulin signaling. Taken together, our findings reveal that GHS-R is an important immunometabolic regulator and governs macrophage polarization through regulating mitochondrial function and cellular metabolism of macrophages, moderating hepatic and systemic inflammation and insulin sensitivity. GHS-R antagonist may present a novel therapeutic strategy for the metabolic disorders such as NAFLD.

Disclosure

D. Kim: None. J. Lee: None. Q. Pan: None. J. Noh: None. H. Wang: None. H. Han: None. C. Wu: None. S. Guo: None. Y. Sun: None.

Funding

American Diabetes Association (1-15- BS-177 to Y.S.); National Institutes of Health
1999-P: Cysteine-Rich Protein 2 (CRIP2) Is a Novel Transcriptional Regulator of Uncoupling Protein 1
Authors: DARCY JUSTIN , WANG CHIH-HAO , LUNDH MORTEN , SHAMSI FARNAZ , LYNES MATTHEW , D'APRIGNY THOMAS , EMANUELLI BRICE , TSENG YU-HUA
Published: June 2019

Since traditional weight loss strategies have been unsuccessful at slowing the increasing rate of obesity, alternative strategies are in high demand. Brown adipose tissue (BAT) is a thermogenic tissue that efficiently expends energy. Thermogenesis in BAT occurs through the action of uncoupling protein 1 (UCP1). Identifying novel transcriptional regulators of UCP1 may lead to potential targets to increase BAT activity since its expression is under tight transcriptional control. Engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) is a modified CRISPR/Cas9 methodology that identifies novel transcriptional regulators. EnChIP uses dCas9 (lacks endonuclease activity) and guide RNAs (gRNAs) targeting a specific promoter region. The DNA-bound dCas9 can be immunoprecipitated, and the associated promoter region/transcriptional machinery can be sent for proteomic analysis to identify novel transcriptional regulators. Following β-adrenergic stimulation, which activates UCP1 gene transcription in brown adipocytes, we performed enChIP by using five gRNAs targeting the UCP1 promoter. EnChIP analysis of the UCP1 promoter revealed cysteine-rich protein 2 (CRIP2) as a putative transcriptional regulator of UCP1, which was validated by ChIP-PCR. We demonstrated that knocking down CRIP2 in brown adipocytes significantly decreased β-adrenergic-induced UCP1 expression and heat production. Mechanistically, we demonstrated that following β-adrenergic stimulation, CRIP2 was phosphorylated and translocated from the cytosol to the nucleus, where it interacted with peroxisomal proliferator-activated receptor gamma (PPARγ). Taken together, using the unbiased CRISPR-based enChIP approach, we identify CRIP2 as a novel transcriptional regulator that modulates UCP1 expression in response to β-adrenergic stimulation. These findings pave the foundation for developing new anti-obesity and antidiabetes therapies.

Disclosure

J. Darcy: None. C. Wang: None. M. Lundh: Employee; Self; Gubra. F. Shamsi: None. M. Lynes: None. T. d'Aprigny: None. B. Emanuelli: None. Y. Tseng: Other Relationship; Self; Chugai Pharmaceutical Co., Ltd.

Funding

National Institutes of Health (R01DK077097, R01DK102898, T32DK007260)
20-OR: GPR142 Agonists Increase Glucose-Dependent Insulin Secretion and Differentially Regulate Hormone Secretion in Preclinical and Phase 1 Studies
Authors: PRATT EDWARD J. , OURA TOMONORI , MA XIAOSU , LIN HUA V. , MA TIANWEI , EFANOV ALEXANDER M. , THOMAS MELISSA K.
Published: June 2019

Agonists of G-protein-coupled receptor 142 (GPR142) are reported to increase insulin secretion and lower glucose in preclinical models. We identified a novel agonist, LY3325656 (LY) that selectively activates GPR142 in humans and mice. In preclinical models, LY increased glucose-dependent insulin secretion (GDIS) and secretion of glucagon and incretin hormones GLP-1 and GIP. To elucidate mechanisms of LY in humans, we designed a multi-part, single-dose study in 24 healthy volunteers (HV) and 12 patients with type 2 diabetes (T2D) (NCT03115099). LY was well tolerated through dose escalation up to 1950 mg in HV, with a median time to peak concentration of 4 hours and half-life of 4.59 hours. A graded glucose infusion evaluated GDIS, and LY demonstrated increased insulin secretion at higher doses (750-1950 mg). A 4-period, crossover design consisting of placebo (Pb), LY (1950mg), LY+DPP-IV inhibitor, and GLP-1R agonist treatment investigated glucose-lowering and incretin pharmacology in patients with T2D. LY significantly reduced post-meal glucose levels (2-hour glucose AUC) by >10% during a mixed meal tolerance test. Increases in post-meal GLP-1 AUC concentrations with LY compared to Pb were seen in a few, but not all patients with T2D. Serum glucagon levels were consistently increased by LY in patients with T2D and HV. We conclude that GPR142 activation in humans increases GDIS and augments glucagon secretion but does not substantively augment GIP or GLP-1 secretion. Our data suggest that acute stimulatory effects of GPR142 on insulin and glucagon secretion translate from preclinical models to man, unlike preclinical effects on gut peptide secretion. Innovative and efficient single-dose studies can provide key mechanistic understanding of the translatability of preclinical findings for the treatment of T2D.

Disclosure

E.J. Pratt: Employee; Self; Eli Lilly and Company. T. Oura: Employee; Self; Eli Lilly and Company. Stock/Shareholder; Self; Eli Lilly and Company. X. Ma: Employee; Self; Eli Lilly and Company. H.V. Lin: Employee; Self; Eli Lilly and Company, Forkhead BioTherapeutics, Inc. Employee; Spouse/Partner; Novo Nordisk A/S. Stock/Shareholder; Self; Eli Lilly and Company, Forkhead BioTherapeutics, Inc. Stock/Shareholder; Spouse/Partner; Novo Nordisk A/S. T. Ma: None. A.M. Efanov: Employee; Self; Eli Lilly and Company. M.K. Thomas: Employee; Self; Eli Lilly and Company. Stock/Shareholder; Self; Eli Lilly and Company.
2003-P: The Role of Re1-Silencing Transcription Factor (REST) in Adipocytes
Authors: FUWA MASAYUKI , KAJITA KAZUO , TAGUCHI KOICHIRO , KITADA YOSHIHIKO , IKEDA TAKAHIDE , ISHIZUKA TATSUO , MORITA HIROYUKI
Published: June 2019

We have reported that several neuronal genes, glutamate receptor 7 and synuclein γ, are expressed in adipocytes. Since the individual roles of these neuronal genes are different, it is unlikely that these genes carry out any common functions together. Numerous neuronal genes are negatively regulated by REST. Therefore, we generated the mice lacking REST in adipocytes to investigate the role of these genes. Rest2lox/2lox mice were mated with Adipoq-Cre mice, and the resultant Adipoq-Cre/Rest2lox/2lox mice were compared with Adipoq-Cre mice. Male Adipoq-Cre/Rest2lox/2lox mice exceeding 10 weeks of age showed increased body weight and fat mass. There was no difference in food consumption, glucose tolerance evaluated by intraperitoneal GTT (ip-GTT) and adipocyte size. However, glucose tolerance was deteriorated in Adipoq-Cre/Rest2lox/2lox mice after treated with high fat diet (HFD) for 8 weeks. The characteristics of gene expression in epididymal fat isolated from Adipoq-Cre/Rest2lox/2lox mice was examined with microarray with Adipoq-Cre/Rest2lox/2lox mice as a control. Significant increased mRNA levels were detected numerous neuronal genes such as Gdap1, Nrxn1, Nrsn1 and Syn1, in Adipoq-Cre/Rest2lox/2lox mice. In addition, expression of several oncogenes, including Srrm3, Lhfpl5, Stmn3 and Chga (a marker of neuro-endocrine tumor such as pheochromocytoma) were increased in these mice. On the other hand, brown adipose tissue (BAT) in Adipoq-Cre/Rest2lox/2lox mice showed white adipose tissue (WAT)-like appearance. These results suggested that REST might regulate adipocyte differentiation and function in both WAT and BAT.

Disclosure

M. Fuwa: None. K. Kajita: None. K. Taguchi: None. Y. Kitada: None. T. Ikeda: None. T. Ishizuka: None. H. Morita: None.
2007-P: Age-Dependent Metabolic Response to Erythropoietin in Mice
Authors: ROGERS HEATHER , NOGUCHI CONSTANCE T.
Published: June 2019

Erythropoietin (EPO), the cytokine required for erythrocyte production, regulates glucose homeostasis and fat mass accumulation in animal models. For example, young (5-6 weeks) mice on high fat diet (HFD) treated with EPO for 5 weeks show a dose dependent increase in hematocrit and reduction in fat mass accumulation/weight gain. To assess potential age and sex dependent EPO metabolic response, C57/Bl6 mice beginning at 3 months, 4 months or 6 months of age were fed normal chow (NC) or high fat diet (HFD) and treated with EPO or saline for up to 6 weeks. After 3 weeks of EPO treatment, all mice exhibited the expected increase in hematocrit (hct up to 70%) that was accompanied by an improvement in glucose tolerance. Body weight gain was unchanged in the 3-month groups, while the 4- and 6-month males on HFD showed a reduced gain in body weight with 3-weeks of EPO treatment. Since earlier studies suggested an increased reduction in fat mass with extended EPO treatment, EPO treatment was continued for three more weeks. At 6 weeks EPO treatment, body weight gain was not affected in the 3-month groups on NC or HFD but was reduced in the 4- and 6-month groups on HFD in male and female mice. These differences in weight gain appear to be due to differences in fat accumulation. However, after 6 weeks EPO treatment, about 66% of the 3-month group, 70% of the 4-month group showed reduced hematocrit below baseline to low as 22% in the 3-month group and 11% in the 4-month group. Mice with the lowest hct showed the greatest impairment in glucose tolerance. In the 6-month group on HFD, EPO treatment for 6 weeks decreased body weight in both males and females. However, hct decreased in all 6-month mice treated with EPO for 6 weeks, falling as low as 15% and the improved glucose tolerance with EPO treatment seen at 3 weeks was no longer apparent. These data suggest an age dependent immune response at 6 weeks of human EPO treatment in mice and production of anti-human EPO antibodies with the potential to also reduce erythropoiesis and glucose tolerance associated with endogenous EPO.

Disclosure

H. Rogers: None. C.T. Noguchi: None.

Funding

National Institutes of Health
2018-P: Simultaneous Monitoring of Circulating Levels of Metabolic-Related Aging Biomarkers in Young and Elderly Mice
Authors: ZENG LING
Published: June 2019

Aging is associated with a decline in lean muscle mass and development of metabolic dysfunction. Skeletal muscle-derived, exercise-induced “myokines” are believed to have anti-inflammatory effects and substantially regulate glucose and fatty acid metabolism. To better understand the linkage between aging associated metabolic dysfunctions and aberrant myokine signaling, we use the MILLIPLEX® map Mouse Myokine Panel and the Mouse Aging Panel (under development) to measure circulating myokine and aging related biomarkers in youth and elderly C57BL/6 mice (Youth: ∼8 weeks old, n=16; Elderly: ∼48 weeks old, n=16). Impaired regenerative capacity and reduction of myokine secretion are hallmarks of aged skeletal muscle. In the elderly group, the myokine SPARC concentrations were 40% that of the youth group and the myogenesis inhibitor GDF8 concentrations were 7-fold higher in the elderly vs. youth. Interestingly, a 21-fold higher level of EPO and a 3-fold higher level of OSTN were observed in the elderly vs. youth groups suggesting a possible compensation effect in these aged mice. We observed the novel type 2 diabetes associated chemokine fractalkine level is 604-fold higher in elderly vs. youth groups. In addition, 41-fold higher IL-6 and 2-fold higher CCL27 in elderly vs. youth coincide with the theory of aberrant glucose metabolism being associated with elevated adipocyte inflammation in elderly populations. Body composition change, and aberrant energy sensing are pervasive features of aging. We found the starvation hormone FGF-21 level was 23-fold higher in elderly mice vs. youth, while the satiety hormone leptin was 3-fold higher. In brief, we measured multiple novel myokines and geriatric related metabolic biomarkers using the MILLIPLEX® map quantitative immunoassay platform. Analytical validation revealed excellent assay performance well suited for biomarker monitoring in mouse models of age-related disease research.

Disclosure

L. Zeng: None.
2024-P: Protective Role of Lipocalin 2 in High-Fat Diet-Induced Gut Microbiota Dysbiosis
Authors: QIU XIAOXUE (SHERYL) , MACCHIETTO MARISSA , SHEN STEVE , CHEN XIAOLI
Published: June 2019

Diet is the most important factor that shapes diversity in gut microbiome. However, what host factors are involved in dietary regulation of gut microbiota remains largely unknown. Lipocalin 2 (Lcn2) is expressed in intestine, and its expression is upregulated during acute intestinal inflammation. Herein, we investigated the role of Lcn2 in shaping gut microbiota during high-fat diet (HFD)-induced obesity. First, we showed that 12 weeks of HFD feeding led to a significant increase in Lcn2 protein levels in ileum and colon, but a decrease in Lcn2 levels in feces of C57BL/6J mice. This suggests that Lcn2 release into the gut lumen decreases following long-term HFD feeding. Second, we revealed a time course of HFD-induced changes in gut microbiota. Feces were collected from wild type (WT) and Lcn2 knockout (LKO) mice at 2, 4, 8 and 16 weeks of HFD feeding for DNA extraction and 16S sequencing. Compared to WT mice, LKO mice displayed a trend towards lower bacterial diversity (p=0.078) at 4 weeks but not 16 weeks of HFD feeding, suggesting that Lcn2 may have a protective role against gut microbiota dysbiosis during short-term HFD consumption. More interestingly, species richness (the number of bacterial species) was increased at 4 and 8 weeks of HFD feeding and then declined 16 weeks after HFD feeding in WT mice; this time-dependent change in the richness was not observed in LKO mice. Principal coordinate analysis indicates a difference in gut microbiota composition between WT and LKO mice when all time points were considered for the analysis. The heatmap shows that the abundance of short-chain fatty acid-producing bacteria such as Lachnospiraceae and Muribaculaceae was significantly decreased in HFD-fed LKO mice. Lastly, upon 16 weeks of HFD feeding, LKO mice had increased levels of STAT3 and NFκB phosphorylation in colon compared to WT mice, suggesting increased intestinal inflammation. We conclude that Lcn2 plays a protective role in HFD-induced intestinal inflammation and gut microbiota dysbiosis.

Disclosure

X. Qiu: None. M. Macchietto: None. S. Shen: None. X. Chen: None.
2039-P: Age-Related Dysfunction of Brown Adipose Progenitor Cells
Authors: GRAJA ANTONIA , GOHLKE SABRINA , SARAIVA LUIS , SCHULZ TIM J.
Published: June 2019

Aging is a primary risk factor for the development of obesity and diabetes mellitus and is accompanied by impaired brown adipocyte formation. Deprivation of thermogenic capacity may contribute to an obesity-prone phenotype of aged individuals. To determine whether the age-related decline of brown adipogenesis is due to a cell-autonomous dysfunction of brown adipogenic cells, we isolated stem cells from brown adipose tissue of mice aged 2.5 or 25 months for characterization. While lipid accumulation capacity was unchanged, expression of brown adipocyte markers, such as Uncoupling protein-1 (Ucp1), were significantly reduced in aged cells after differentiation, suggesting that a cell-intrinsic defect is responsible for impaired brown adipogenesis during aging. We subsequently generated immortalized single stem cell clones from both age groups, which displayed a similar age-related defect. Transcriptomes generated from a total of 126 clones show that brown fat-derived stem cells from old mice more closely resemble stem cells isolated from white adipose tissue. Among the genetic pathways significantly altered in aged stem cells are changes to extracellular matrix function as well as cell-intrinsic pathways. Importantly, aged stem cells are characterized by an age-dependent induction of cellular senescence with increased expression of cell cycle inhibitors p16, p21 and p53. To further test the link between senescence and reduced brown adipogenic potential, we isolated brown adipogenic stem cells from a mouse model with conditional inactivation of p53. Confirming our hypothesis, p53-deficient stem cells displayed a marked increase in Ucp1-expression upon differentiation when compared to wild type stem cells. These data taken together suggest that aging of brown adipogenic stem cells may contribute to loss of brown adipocytes and that inhibition of cell cycle blockage may restore their ability to undergo full brown adipogenic differentiation.

Disclosure

A. Graja: None. S. Gohlke: None. L. Saraiva: None. T.J. Schulz: None.

Funding

German Center for Diabetes Research; German Research Foundation
204-OR: Advancing Our Understanding of the Relationships between Diabetes Distress and Depressive Symptoms with Biobehavioral Variables in Adolescents with Diabetes
Authors: ABUJARADEH HIBA , WONG JESSIE J. , HOOD KOREY K.
Published: June 2019

In adolescents with diabetes, associations of depressive symptoms and diabetes distress (DD) with self-care behaviors and glycemic control vary considerably across studies, yet in adults, DD is a stronger predictor than depressive symptoms. This secondary data analysis compared the strength of the association of DD and depressive symptoms on bio-behavioral (HbA1c and average daily blood glucose monitoring checks [BGM]) and perceived global health among adolescents with diabetes.

Results: Data from 119 adolescents (75% T1D; diabetes duration of 4.8±4.5 years; HbA1c 8.7 ±2.3%; 62% female; mean age of 15.8±2.1 years; 36% Hispanic, 31% white) screened in clinic were analyzed. Teens completed the 2-item DD Scale (DDS2), the PHQ-9, and PROMIS Pediatric Global Health (PGH). Demographics, HbA1c, and BGM were obtained from medical chart review. Of the 119, 6% had severe depression (PHQ >10) and 22% had moderate to severe DD. DD was significantly correlated with HbA1c and BGM (r=.27, -.32;p=.004, .002; respectively); and both DD and PHQ were significantly correlated with the PGH (r=.43, -.57;ps<.001; respectively). In the regression models that included both DD and PHQ along with demographic and clinical covariates, DD was a significant predictor of HbA1c (ß= .31, partial r=.27, p=.004) and BGM (ß= -.24, partial r=-.23, p=.03). PHQ was not a significant predictor of either, but was a significant predictor of PGH (ß=-.6, partial r=-.47, p<.001). Type of diabetes was not a significant covariate.

Conclusion: DD had stronger associations with diabetes biobehavioral variables than depressive symptoms. Considering the outcomes are diabetes-specific, this is not surprising. However, it shows that simply screening for depression may miss an important aspect of the psychosocial health of adolescents with diabetes. Appropriate intervention should follow positive screens of DD to improve adolescents’ diabetes-related variables.

Disclosure

H. Abujaradeh: None. J.J. Wong: None. K.K. Hood: Consultant; Self; Lilly Diabetes. Research Support; Self; Dexcom, Inc. Speaker's Bureau; Self; Johnson & Johnson Diabetes Institute.
2045-P: Lipid-Induced Mitochondrial Dysfunction Interferes with Retinoic Acid Signaling in Hepatocytes
Authors: KARMAKAR ESHANI , DAS NABANITA , MUKHOPADHYAY SATINATH , ROY SIB SANKAR
Published: June 2019

Insulin Resistance (IR) plays a key role in pathogenesis of type 2 diabetes. NAFLD, considered by some as the hepatic manifestation of IR, leads to NASH, cirrhosis and rarely hepatocellular carcinoma. Hepatic Cirrhosis secondary to NAFLD is an emerging public health tsunami. IR is known to impair lipid induced hepatic Retinoid metabolism. Lecithin: Retinol Acyl Transferase (Lrat) plays a crucial role in retinoid homeostasis. Studies have shown the involvement of all-trans Retinoic Acid (atRA) in mitochondrial biogenesis; it functions mainly through PPAR-α and RXR. We identified overexpression of LRAT in liver of HFD-fed rats, hepG2 and huh7 cell lines. Lipid (Palmitic acid-PA) induced over expression of Lrat was associated with a significant reduction in RA production; thereby impairing PPAR-α mediated fatty acid oxidation (FAO) and fat accumulation in hepatocytes. PA induced overexpression of hepatic Lrat was accompanied by excess lipid influx into the mitochondria resulting in impaired mitochondrial biogenesis and bioenergetics, characterized by loss of membrane potential, elevated ROS generation and impaired FAO. Knockdown of Lrat gene restored reduced levels of PPARα resulting from overexpression of Lrat upon PA treatment. Silencing of hepatic Lrat abrogated neutral fat accumulation in PA treated liver tissue and cell lines. Lipid induced mitochondrial dysfunction and resulting elevation in ROS levels was associated with increased mitochondrial fragmentation and decreased membrane potential; Lrat silencing reversed this process. Taken together our study unravels a novel role of PA induced Lrat overexpression in neutral fat accumulation in the liver that may potentially lead to development of NAFLD and NASH in the short term and cirrhosis and rarely, HCC in the long term. This brings to the fore Lrat silencing as a future drug target to prevent and/or treat fat accumulation in the liver in setting of Insulin Resistance.

Disclosure

E. Karmakar: None. N. Das: None. S. Mukhopadhyay: None. S. Roy: None.
205-LB: Use of Internet and Mobile Technology for Health Communications in the Jackson Heart Study
Authors: ANUGU PRAMOD R. , VALLE KAREN A. , WINTERS KAREN , CORREA ADOLFO
Published: June 2019

The availability of internet and/or mobile phone technology (IMT) provides an opportunity to explore more practical approaches to collect and disseminate information related to diabetes prevention and control in cohort studies. However, the extent of use of IMT in cohort studies of high-risk populations is unclear. In the Jackson Heart Study (JHS), a community-based cohort study of cardiovascular disease among African Americans with an increased prevalence of diabetes, we evaluated the prevalence and demographic correlates of IMT use. As part of annual follow-up telephone interviews of JHS participants in 2017-2018, we conducted a survey on IMT use and examined prevalence and demographic correlates of IMT use. Of 3,810 JHS participants in annual follow-up interviews, 2,564 (67.3%) completed the IMT use survey, of whom 2,262 (88.2%) reported IMT use. Compared to non-users, IMT users were more likely to be men (37% vs. 26%), younger (mean age 70 vs. 81years), and to have attained high school or more years of education (89% vs. 57%) and middle-affluent income status (91% vs.78%) (Table 1). For a majority of JHS participants, it may be feasible to use IMT to conduct intensive interviews and pilot diabetes-related communications. For participants with more limited use of IMT, standard methods will continue to be needed and novel methods developed to capture and disseminate information on diabetes prevention and control.

Disclosure

P.R. Anugu: None. K.A. Valle: None. K. Winters: None. A. Correa: None.

Funding

Jackson State University (HHSN268201800013I); Tougaloo College (HHSN268201800014I); Mississippi State Department of Health (HHSN268201800015I); University of Mississippi Medical Center (HHSN268201800010I, HHSN268201800011I, HHSN268201800012I); National Heart, Lung, and Blood Institute; National Institute for Minority Health and Health Disparities
2054-P: Endothelial Dysfunction, Inflammation, and Oxidative Stress Urinary Biomarkers in Obese Children from Alabama
Authors: SELVARAJU VAITHINATHAN , AYINE PRISCILLA , PARRA EMILY P. , BROWN MICHAEL , JEGANATHAN RAMESH B. , THANGIAH GEETHA
Published: June 2019

Obesity is a state of chronic low-level inflammation closely associated with the oxidative stress. Childhood obesity has been related to endothelial dysfunction, inflammation and oxidative stress markers individually. The aim of this study was to determine the association of biomarkers of the inflammation, oxidative stress and endothelial dysfunction in urine samples of healthy and obese children. Thirty healthy and thirty overweight/obese elementary school children aged between 6 to 10 years from Alabama participated in this study. Anthropometric measurements were measured using WHO recommendations. Urine samples were analyzed for biomarkers of low-grade inflammation such as interleukin-6 (IL-6), C-reactive protein (CRP), α-1-acid glycoprotein (AGP); oxidative stress such as 8-isoprostane, 8-hydroxy-2'-deoxyguanosine (8-OHdG); and endothelin-1 (ET-1). Linear regression analysis was performed to analyze the association between the parameters. The overweight/obese children have significantly increased levels of IL-6, CRP, ET1 and 8-isoprostane compared to normal weight. CRP, IL-6, AGP and 8-isoprostane have significant positive association with at least one of the obesity measures except the ET-1 and 8-OHdG. ET-1 is significantly positively correlated with IL-6 and 8-isoprostane. The increased urinary IL-6 and 8-isoprostane can be used as a non-invasive benchmark to monitor childhood obesity.

Disclosure

V. Selvaraju: None. P. Ayine: None. E.P. Parra: None. M. Brown: None. R.B. Jeganathan: None. G. Thangiah: None.

Funding

National Institute of Food and Agriculture
2057-P: HDL-Cholesterol Concentration Independently Predicts Future Accumulation of Intra-abdominal Fat in Japanese Americans
Authors: SONG SUN OK , HWANG YOU CHEOL , KAHN STEVEN E. , LEONETTI DONNA L. , FUJIMOTO WILFRED Y. , BOYKO EDWARD J.
Published: June 2019

Background: Both intra-abdominal fat (IAF) and HDL cholesterol (HDL-C) are known to be associated with cardiometabolic health problems, with an inverse correlation between them. However, there is little information about the association between HDL-C concentration with future accumulation of IAF. We conducted a prospective study to evaluate whether baseline HDL-C concentration was associated with the accumulation of CT-measured intra-abdominal fat (IAF) over 5 years.

Methods: All participants were Japanese-American men and women between the ages of 34 to 74 years. HDL-C concentration and CT-measurements of IAF, abdominal (SCF) and thigh (TF) subcutaneous fat cross-sectional areas were evaluated at baseline and 5 year follow-up visits. Data were analyzed to determine the association between HDL-C and future 5-year accumulation of IAF.

Results: A total of 545 subjects with and without diabetes (296 men, 249 women, and mean age 53.6±11.4 and 54.0±12.0 years, respectively) were included. The mean (SD) HDL-C concentration was 1.3 mmol/L (0.4) in men and 1.7 mmol/L (0.4) in women and mean (SD) IAF was 102 cm2 (53.8) in men and 74.5 cm2 (46.8) in women. Baseline HDL-C concentration was inversely associated with Δ IAF over 5 years using multivariable regression analysis adjusting for age, sex, diabetes family history, weight change over 5 years, and baseline measurements of BMI, IAF, abdominal SCF, abdominal circumference, TF, and HOMA-IR.

Conclusions: These results demonstrate that HDL-C concentration predicts significantly future accumulation of IAF over 5 years independent of age, sex, insulin sensitivity, and body composition in nondiabetic Japanese-American men and women.

Disclosure

S. Song: None. Y. Hwang: None. S.E. Kahn: Advisory Panel; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Eli Lilly and Company, Intarcia Therapeutics, Inc., Janssen Pharmaceuticals, Inc., Merck & Co., Inc., Novo Nordisk A/S. Consultant; Self; Neurimmune. Other Relationship; Self; Boehringer Ingelheim Pharmaceuticals, Inc., Merck & Co., Inc., Novo Nordisk A/S. D.L. Leonetti: None. W.Y. Fujimoto: None. E.J. Boyko: None.

Funding

National Institutes of Health DK31170, HL49293); University of Washington (DK017047, DK035816, RR-000037); VA Puget Sound Health Care System (to E.J.B., S.E.K.)
2065-P: Effects of Obesity on Productivity in the Australian Population: A Life Table Modelling Study
Authors: MENON KIRTHI ARAVIND , ZOMER ELLA , LIEW DANNY , COURTEN BARBORA DE
Published: June 2019

Background: The prevalence of obesity has tripled over the last 40 years, which represents a growing public health concern. Other than exerting a major health impact, the economic consequences of obesity are also significant. The cost of obesity in Australia in 2010 was estimated to be close to AUD $57 billion, including both direct and indirect costs. Absenteeism and presenteeism result in reduced or loss of work productivity, which significantly impacts the economy. The present study sought to estimate the loss of productivity due to obesity over the next 10 years among Australians.

Methods: Prevalence, incidence, mortality, and absenteeism data were obtained from published sources. Life table models were created to estimate the productivity of Australians aged 17-69 years, with follow-up for 10 years or until age 70 years. Productivity was measured in terms of productivity adjusted life years (PALYs) lived. The life table was constructed for the current cohort of Australians of working age with obesity (including incident cases) and then repeated for the same cohort assuming they were not obese. The differences in outcomes between the two modelled analyses reflected the impact of obesity on productivity. All outcomes were discounted by 5% per annum.

Results: Obesity resulted in over 54,000 excess deaths, and a loss of 285,000 (discounted) PALYs. If the economic value of each PALY was assumed to be equivalent to the Australian gross domestic product per equivalent full-time worker, which in 2015 was AUD $192,000, then the loss of PALYs attributable to obesity would equate to a loss of AUD $55 billion to the Australian economy.

Conclusions: The results support the need to implement measures, strategies and ultimately policies to manage and control the growing epidemic of obesity.

Disclosure

K. Aravind Menon: None. E. Zomer: Consultant; Self; Amgen Inc., AstraZeneca, Pfizer Inc., Shire. D. Liew: Advisory Panel; Self; AstraZeneca, Bayer AG. Research Support; Self; AbbVie Inc., AstraZeneca, Bristol-Myers Squibb Company, CSL Behring, Pfizer Inc. B. de Courten: None.
2083-P: Monogenic Obesity Mutations Lead to Less Weight Loss after Bariatric Surgery: A 6-Year Follow-Up Study
Authors: HU CHENG
Published: June 2019

Objectives: Bariatric surgery is emerging as the most effective treatment option for patients with obesity. Hypothalamic arcuate nucleus plays an important role in metabolic homeostasis. However, the influence of mutations related to the feeding center on weight loss after bariatric surgery is still unclear. We aimed to diagnose monogenic obesity by whole-exome sequencing (WES) and explore whether monogenic mutations influence the effectiveness of bariatric surgery.

Methods: We collected obese patients aged 15 to 55 with a BMI >28 kg/m2 and who underwent laparoscopic sleeve gastrectomy from March 2011 to June 2017 in Shanghai. Data related to weight loss and metabolic characteristics preoperatively and postoperatively were collected, including fasting blood glucose (FBG), high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol and triglycerides. WES was performed in obese patients using genomic DNA from whole blood samples.

Results: We investigated the proportion of 131 obese adults with one mutation as high as to 8.4% and then evaluated the association between these mutations and weight loss. Mutation carriers had less weight loss over both short-term and long-term periods. Survival analyses indicated it was harder to attain the goal of 20% weight loss for mutation carriers (Plog-rank=0.001; Pbreslow<0.001), and the difference remained significant with a Cox regression model. Improvement in FBG, HDL cholesterol and triglycerides postoperatively were observed in both groups, while there were significant differences between the two groups.

Conclusions: Our data indicated that 8.4% of obesity cases were caused by change in genetics, and mutations had negative effects on the efficacy of bariatric surgery.

Disclosure

C. Hu: None.
2087-P: Association between L-FABP with Lipid Metabolism and Alteration after Laparoscopic Sleeve Gastrectomy in Chinese Obese Subjects
Authors: YOU HUI , LIN ZIWEI , ZHU CUILING , WEN XIN , ZHANG MANNA , QU SHEN
Published: June 2019

Liver fatty acid-binding protein (FABP1) is associated with intracellular transportation of fatty acids. However, only few studies indicated the relationship between serum FABP1 and metabolism. This study aimed to investigate the association of FABP1 levels and metabolism factors, and its change with laparoscopic sleeve gastrectomy (LSG) in Chinese. The cross-sectional analysis included a total of 187 subjects, of which 65 were with normal weight (BMI 18-24 kg/m2), 59 with over weight (BMI25-27 kg/m2), and 64 with obesity (BMI≥28 kg/m2). We found FABP1 levels in obese (303±235pg/ml, p<0.05) and over-weight (189±187pg/ml, p<0.07) subjects were significantly higher than those in normal-weight subjects (142±93pg/ml), and higher in NAFLD (225±220pg/ml, p< 0.01) than in those without (129±74pg/ml) as well. In order to further investigated the correlation between FABP1 and metabolic factors, we performed a linear-regression analysis. We found FABP1 was positively correlated with BMI (R2=0.201, p<0.001), ALT(R2=0.324, p<0.001), AST(R2=0.387, p<0.001), m-AST(R2=0.160, p<0.001),[[γ]]-GT(R2=0.106, p<0.001), DBIL(R2=0.078, p= 0.001), UA(R2 =0.111, p<0.001), FBG(R2=0.066, p<0.001), LDL(R2 =0.042, p=0.005) and negatively correlated with HDL(R2 =0.051, p= 0.002). Moreover, we also enrolled 25 severely obese patients who underwent LSG to observe its effects on FABP1. We found with the decline of BMI after surgery, FABP1(294±211pg/ml) were also gradient decreased at 3-month (230±227pg/ml, p=0.463), 6-month (144±154pg/ml, p=0.09) and 12-month (115±32pg/ml, p=0.045).

In conclusion, serum FABP1 were correlated with obesity and NAFLD, and it might be an important mediator participating in metabolism disorders.

Disclosure

H. You: None. Z. Lin: None. C. Zhu: None. X. Wen: None. M. Zhang: None. S. Qu: None.

Funding

National Key Research and Development Program of China (2018YFC1314101); National Natural Science Foundation of China (81700752, 81500687)
2094-P: Decreased Human Adipose Tissue-Resident Eosinophils in Obese Subjects Is Associated with Increased Insulin Resistance
Authors: HERNANDEZ JAMES D. , LI TING , RAU CASSANDRA , MASUDA MIA Y. , MADURA JAMES A. , JACOBSEN ELIZABETH A. , DEFILIPPIS ELEANNA
Published: June 2019

Eosinophils (EOS) have been shown to play a major role in animal adipose tissue (AT) metabolism by promoting insulin sensitivity (IS) through an increase in alternatively activated macrophages (M2). Additionally, AT EOS are known to be decreased in obese mice. Currently, translational evidence is lacking for the role of EOS in human AT metabolism and IS. We investigated whether human AT-resident EOS numbers, as in animal models, are decreased in obese subjects and correlate with their degree of insulin resistance (IR). To date, we have studied 16 lean subjects (F15/M1, age 43 +2 years old; BMI 22.5+0.4kg/m2; HOMA-IR: 0.99 +0.38) and 22 obese, IR subjects (17F/5M, age 42 +1.3 years old; BMI: 38.9 +1.0kg/m2; HOMA-IR:5.0 +1.6) undergoing elective laparoscopic surgery. During surgical procedure, subcutaneous (SC) and omental (OM) fat was harvested. Stromal vascular fraction (SVF) was isolated after collagenase digestion of the fat samples. SVF was subjected to FACS analysis using BD Biosciences FACSAria III Cell Sorter with EOS count normalized as percent of CD45+ cells. Viable EOS were isolated and cytospin preparation was stained with a Diff-Quik. EOS isolated from SC AT of lean(L) subjects were found to significantly outnumber those obtained from obese (Ob) subjects by a 2.1:1 ratio (SC L:2.34 +0.29 vs. SC Ob: 1.11 +0.21, P<0.03). A similar, but non-significant, trend was observed in OM. Furthermore, the number of SC AT-EOS was significantly inversely correlated with BMI, % fat and QUICKI index. qPCR analysis of SC AT mRNA revealed significantly increased classically activated macrophage markers (M1) (IL-1, IL-6, TNFα) in Ob vs. L, respectively while M2 markers (ARG-1, CCL24, CCL26) were significantly decreased in Ob vs. L. Based on these results we propose that, as in animal models, AT-resident EOS mediate AT metabolism and potentially IS by preserving an anti-inflammatory milieu within lean AT. Further studies are needed to elucidate the mechanism(s) involved.

Disclosure

J.D. Hernandez: None. T. Li: None. C. Rau: None. M.Y. Masuda: None. J.A. Madura: None. E.A. Jacobsen: None. E. DeFilippis: None.

Funding

Arizona Department of Health Services (14-00003606); Arizona Biomedical Research Commission; Kathryn H. and Roger Penske Career Development Award