bluuebunny/crossref_metadata_2025_split huggingface dataset:
Loss of Fractalkine-CX3CR1 Signaling Exacerbates Obesity-Induced Adipose Tissue Inflammation and Insulin Resistance through M1 Dominant Shift in Macrophages
Authors: NAGASHIMADA MAYUMI , NI YINHUA , OTA TSUGUHITO 
Published: June 2018

Adipose tissue macrophages (ATMs) accumulation through CCR2 and its ligand MCP-1 is considered pivotal in the development of insulin resistance. Recently, we found that CX3C chemokine, fractalkine (CX3cl1) mRNA expression was persistently downregulated in epididymal white adipose tissue (eWAT) of HF diet (HFD)-induced obese (DIO) mice, as compared to lean controls. Furthermore, we demonstrated that loss of fractalkine receptor, CX3CR1, exacerbated HFD-induced insulin resistance and glucose intolerance. Here, we show that fractalkine-CX3CR1 signaling plays a crucial role in the inflammatory response to HF feeding by regulating macrophage M1/M2 status. To determine the effect of fractalkine-CX3CR1 signaling on ATM subsets, we performed flow cytometry analysis to quantify M1/M2 ATMs in eWAT of CX3CR1-/- (KO) and WT mice. Immunofluorescence study of eWAT in DIO mice revealed that CX3CR1 was predominantly expressed by F4/80+ macrophages in crown-like structures (CLS). On an HFD, KO mice had increased macrophage infiltration and formation of CLS in eWAT compared with those of WT mice, despite that weight and adipocyte size were similar. KO mice had 40% or 39% more CD11c+CD206- (M1) ATMs on normal chow (NC) or HFD, and 17% or 49% fewer CD11c-CD206+ (M2) ATMs on NC or HFD, respectively, than WT mice, resulting in predominance of the M1 over M2 ATM population. Furthermore, the predominance of the Ly6Chi over the Ly6C- monocyte population was observed in the peripheral blood and bone marrow (BM) of KO mice on NC or HFD. Importantly, HFD-induced adipose inflammation and hyperinsulinemia were aggravated in chimeric mice lacking CX3CR1 only in myeloid cells (BM transplant of KO to WT) as compared to control mice (BM transplant of WT to WT). Thus, loss of fractalkine-CX3CR1 signaling causes M1 dominant phenotypic shift in ATMs, which contributes to exacerbation of obesity-induced adipose tissue inflammation and insulin resistance.

Disclosure
M. Nagashimada: None. Y. Ni: None. T. Ota: None.
Glycolytic Activation Promotes PDGF-B Production in Adipose Tissue Macrophages during Obesity Development
Authors: ONOGI YASUHIRO , WADA TSUTOMU , MATSUZAWA TAKATOSHI , OKEKAWA AKIRA , WATANABE ERI , TSUNEKI HIROSHI , SASAOKA TOSHIYASU 
Published: June 2018

The vascular network develops with remodeling in adipose tissue (AT) during obesity. We have found that PDGF-B detaches pericytes from the vasculature, which plays a crucial role in angiogenesis during obesity. The expression of PDGF-B was significantly elevated in obesity-associated infiltrated CD11c-positive (+) AT macrophages (ATMs). Interestingly, the expression was low in both CD11c+ ATMs and CD206+ ATMs of lean mice. Since glycolytic and oxidative metabolism are key metabolic programs that regulate ATMs’ polarity and function, we hypothesized that hyperglycemia and inflammation synergistically drive PDGF-B production by reprogramming metabolic pathways in ATMs.
The expression of PDGF-B was increased in the AT of high-fat diet (HFD)-fed mice and STZ-induced hyperglycemic mice. In accordance, exposure to high glucose led to increase the expression of PDGF-B in association with that of Itgax (CD11c) in RAW264.7 macrophages. In peritoneal macrophages, LPS but not IL-4 provoked PDGF-B expression, which was completely blocked by pretreatment with 2-DG, a glycolysis inhibitor. Moreover, LPS effectively augmented high glucose-induced its expression, and that was abolished by MG132, an NFκB inhibitor. Since macrophages highly accumulated in obese AT, where pericytes were detached from vasculature, we examined directly whether PDGF-B from macrophage involves in AT expansion during obesity. To this end, we intraperitoneally injected liposome-encapsulated clodronate (CL) to mice twice a week for 6 weeks during HFD feeding. ATMs were effectively depleted by CL with reduced PDGF-B expression in AT of HFD-induced obese mice. Importantly, HFD-induced obesity was effectively prevented by CL.
In conclusion, PDGF-B from ATMs has a crucial function in AT expansion during obesity. Hyperglycemia and chronic inflammation reprogram metabolic pathways toward glycolysis, and subsequent activation of NFκB plays a pivotal role in the production of PDGF-B in ATMs during obesity.

Disclosure
Y. Onogi: None. T. Wada: Research Support; Self; Sanwa Kagaku Kenkyusho Co., Ltd.. T. Matsuzawa: None. A. Okekawa: None. E. Watanabe: None. H. Tsuneki: None. T. Sasaoka: None.
Deficiency of Melanocortin 4 Receptor Promotes Vascular Vulnerability in Mice
Authors: MORI KENTARO , TSUCHIYA KYOICHIRO , NAKAMURA SUGURU , MIYACHI YASUTAKA , SHIBA KUMIKO , OGAWA YOSHIHIRO , KITAMURA KENICHIRO 
Published: June 2018

Background: Obesity is known as a risk factor of hypertension and abdominal aortic aneurysm (AAA) in humans. Melanocortin 4 receptor (MC4R) is primarily expressed in brain, and mediates most of the anorectic effects of leptin; in humans, deficiency or mutation of MC4R is the most common monogenic form of obesity. However, it remains unknown whether MC4R signaling affects the development of obesity-associated hypertension, vascular vulnerability and aortic aneurysm.
Methods and Results: MC4R-knockout (KO) mice fed a high-fat/high-sucrose diet (HF/HS) for 20 weeks displayed obesity and insulin resistance associated with hyperleptinemia compared to HF/HS-fed wild type mice. HF/HS-fed MC4R-KO mice exhibited hypertension, and showed increase of medial thickness and elastic lamina destruction in aortae. In aortae of HF/HS-fed MC4R-KO mice, genes of MCP-1, TNF-α, and F4/80 were markedly upregulated relative to HF/HS-fed wild type mice. Following the administration of angiotensin II (Ang II, 500 ng/kg/min), HF/HS-fed MC4R-KO mice showed significantly lower survival rate and higher incidence of AAA than HF/HS-fed wild type mice, with increased expression of F4/80- and osteopontin-positive cells in aortae. Furthermore, HF/HS-fed ob/ob mice exhibited significantly lower incidence of Ang II-induced AAA with a trend of higher survival rate than HF/HS-fed MC4R-KO mice. Leptin (100 ng/ml) significantly upregulated osteopontin gene expression in primary cultured vascular smooth muscle cells (VSMCs), but not in primary cultured macrophages.
Conclusion: Our observation clearly demonstrates that deficiency of melanocortin 4 receptor promotes the development of hypertension, vascular vulnerability, and Ang II-induced AAA in mice. It also suggests that hyperleptinemia caused by central leptin resistance could contribute to the development of these vascular phenotypes in MC4R-KO mice, at least partly via induction of osteopontin in VSMCs.

Disclosure
K. Mori: None. K. Tsuchiya: None. S. Nakamura: None. Y. Miyachi: None. K. Shiba: None. Y. Ogawa: Research Support; Self; AstraZeneca. Advisory Panel; Self; Gilead Sciences, Inc.. Research Support; Self; Japan Agency for Medical Research and Development, Japan Society for the Promotion of Science, Mitsubishi Tanabe Pharma Corporation, Mochida Pharmaceutical Co., Ltd., Sumitomo Dainippon Pharma Co., Ltd.. K. Kitamura: None.
Continuous Subcutaneous Delivery of a Glucagon Analog, ICA6150349, to Rats and Nonhuman Primates Demonstrates Safety and Tolerability in a 14-Day Toxicity Study
Authors: ZANE DORIS , SINGH ROSHNI , SHELTON JOSEPHINE , ROLLER SHANE , PAULIK MARK , WAY JAMES , BLACKWELL WILLIAM C. , SRIVASTAVA VED , HODGE REBECCA , FELDMAN PAUL L. , ALESSI THOMAS R. 
Published: June 2018

ICA6150349, a 38-amino-acid peptide, is a glucagon analog that is highly selective for the human glucagon receptor. In a 14-day toxicity study, rats and cynomolgus monkeys received ICA6150349 at doses of 0, 0.875, 8.75, 87.5 mcg/day and 0, 17.92, 179.2, 3584 mcg/day, respectively. Animals were evaluated for reversibility of findings for an additional 14 days. The TK characteristics of ICA6150349 were also determined. In rats, there was a pharmacologically expected dose-dependent decrease in body weight (BW) gains and food consumption, which demonstrated a recovery trend. Clinical pathology and anatomic pathological changes, largely restricted to the exaggerated pharmacological effect of glucagon at high exposures, had mostly recovered during the dose-free period. ICA6150349 exposures (AUC0-24) increased dose-proportionally in male and female rats on D2. On D14, exposure was 40-60% lower than on D2 in the 0.875 and 8.75 mcg/day dose groups and similar to D2 in the 87.5 mcg/day dose group after adjustment for changes in BW. ADAs were observed in rats from all dose groups, but did not impact the TK of ICA6150349. In cynomolgus monkeys, the effects of ICA6150349 were limited to minimally decreased RBC mass (hemoglobin, red blood cell count, and hematocrit) at 3584 mcg/day with recovery. ICA6150349 exposure (AUC0-24) increased dose-proportionally on D2. On Day 14, exposures were 14-65% lower than on D2 in all groups except the 3584 mcg/day females. ADA were observed in 46.7% of treated monkeys; however, ADA status only affected TK at the 3584 mcg/day dose level. Overall, ICA6150349 was well tolerated. Nonclinical safety studies identified no adverse effects (based on the minimal magnitude of change, severity grading and reversibility) at any dose level tested with a NOAEL of 87.5 mcg/day in rats and 3584 mcg/day in monkeys. All observed effects have been non-adverse, reversible, and consistent with known actions of glucagon.

Disclosure
D. Zane: None. R. Singh: None. J. Shelton: None. S. Roller: Employee; Self; Intarcia Therapeutics, Inc. M. Paulik: Employee; Self; Intarcia Therapeutics, Inc. J. Way: Employee; Self; Intarcia Therapeutics, Inc. W.C. Blackwell: Employee; Self; Intarcia Therapeutics, Inc. V. Srivastava: Employee; Self; Intarcia Therapeutics, Inc. R. Hodge: Employee; Self; Intarcia Therapeutics, Inc.. Stock/Shareholder; Self; Intarcia Therapeutics, Inc. P.L. Feldman: Employee; Self; Intarcia Therapeutics, Inc. T.R. Alessi: Employee; Self; Intarcia Therapeutics, Inc..
Cost-Related Insulin Underuse Is Common and Associated with Poor Glycemic Control
Authors: HERKERT DARBY M. , VIJAYAKUMAR PAVITHRA , LUO JING , SCHWARTZ JEREMY , RABIN TRACY L. , DEFILIPPO EUNICE M. , LIPSKA KASIA J. 
Published: June 2018

Insulin is an essential medicine for people with diabetes (DM), but increasing prices have threatened its affordability. We examined the prevalence of and factors associated with cost-related insulin underuse.
We administered a cross-sectional survey to patients with DM prescribed insulin at Yale Diabetes Center (YDC). Our primary outcome was cost-related underuse in the past 12 months, defined by a positive response to any 1 of 6 questions: Did you ⋯ 1) Use less insulin than prescribed 2) Try to stretch out your insulin 3) Take smaller doses of insulin than prescribed 4) Stop insulin 5) Not fill an insulin prescription 6) Not start insulin ⋯ because of cost? We examined the association of cost-related underuse with HbA1c &gt;9% using logistic regression controlling for age, sex, age, DM duration, and income.
Out of 354 patients prescribed insulin who had YDC visit in July 2017, 199 (56.2%) completed the survey (50.8% female, 60.8% white, 41.7% type 1). Of these patients, 51 (25.5%) reported cost-related insulin underuse. Patients with cost-related underuse had lower income levels, variable drug coverage and employment (Figure), and 3-fold higher odds of HbA1c &gt;9% (p = 0.03) than patients who did not report underuse.
One in four patients at our urban diabetes center reported cost-related insulin underuse, and this was associated with poor glycemic control. These results highlight an urgent need to address high insulin prices in the U.S.

Disclosure
D.M. Herkert: None. P. Vijayakumar: None. J. Luo: Consultant; Self; Alosa Health, Inc.. J. Schwartz: None. T.L. Rabin: None. E.M. DeFilippo: None. K.J. Lipska: Other Relationship; Self; Centers for Medicare and Medicaid Services. Research Support; Self; National Institutes of Health.
Small Proliferative Adipocytes, Newly Identified Adipose Progenitor, as the Precursors of Beige Cells
Authors: TAGUCHI KOICHIRO , KAJITA KAZUO , FUWA MASAYUKI , ASANO MOTOCHIKA , KITADA YOSHIHIKO , IKEDA TAKAHIDE , ISHIZUKA TATSUO , MORITA HIROYUKI 
Published: June 2018

We previously identified proliferative cells in adipose tissue expressing adipocyte specific genes and preadipocyte specific genes, which were named small proliferative adipocytes (SPA). Furthermore, we found that SPA specifically express proliferin (PLF) and have superior potential for adipogenic differentiation as compared to stromal vascular cells (SVC). In this study, we investigated the role of SPA. A comparison between differentiated SPA (D-SPA) and differentiated SVC (D-SVC) revealed that UCP1 and PLF were strongly expressed in D-SPA. Therefore, we speculated that SPA might be progenitor of beige cells. The real time PCR analyses and the immunohistochemical studies revealed that beige cells associated genes, including UCP1, PPARα and PGC1α were significantly increased in D-SPA. Treatment with pioglitazone and CL316234, a specific β3-adrenergic receptor agonist, converted cultured SPA into cell clusters like beige cells which expressed UCP1 and activation of mitochondrial function confirmed by MitoRed stain. Furthermore, PLF was expressed in beige cells induced by continuous injection of CL316243.
In conclusion, since D-SPA were found to express UCP1 and PLF, which were detected in beige cells, but not WAT, SPA were considered to be a progenitor of beige cells. There might be two lineages of cells in adipose tissue; adipose progenitor cells in SVC and WAT (PLF-), and SPA and beige cells (PLF+). The combination of pioglitazone and CL316423 produced an additive effect of inducing browning in SPA.

Disclosure
K. Taguchi: None. K. Kajita: None. M. Fuwa: None. M. Asano: None. Y. Kitada: None. T. Ikeda: None. T. Ishizuka: None. H. Morita: None.
The SDF1-CXCR4 Signals Regulate Adipose Tissue Expansion by Modulating Angiogenesis in Diet-Induced Obesity in Mice
Authors: WADA TSUTOMU , WATANABE ERI , KOTERA YURI , ONOGI YASUHIRO , TSUNEKI HIROSHI , SASAOKA TOSHIYASU 
Published: June 2018

Adipose tissue (AT) expands with angiogenesis in obesity. Mature vessels in the epididymal white adipose tissue (eWAT) are tightly wrapped with pericytes (PCs), which attenuates endothelial (ECs) proliferation. We found that obesity-associated increase of PDGF-B promoted detachment of PCs from nature vessels; thereby vessels were prone to neoangiogenesis. In this study, we aimed to clarify the role of stromal cell derived factor 1 (SDF1), another proangiogenic factor in the AT development in obesity. In the whole-mount immunofluorescence study of organ cultured eWAT, treatment with PDGF-B or SDF1 for 24 h significantly promoted detachment of PCs from vessels. The effect of SDF1 on PCs was reproduced by treatment with ATI2341, a potent CXCR4 agonist, but not by VUX11207, a CXCR7 agonist. Interestingly, PCs were well associated with vessels when tissue was treated with both PDGF-B and SDF1. In addition, the association was declined by preincubation with AMD3100, a CXCR4 antagonist or when AT was derived from Pdgfrb knockout mice. To further explore the in vivo effect of SDF1 in AT during obesity, we administrated Anagliptin (Ana), a DPP-4 inhibitor to C57BL/6J mice fed chow or HFD for 12 weeks, since SDF1 is a known substrate for DPP-4. The protein amount of SDF1 was significantly elevated in the eWAT of Ana-treated mice. Importantly, HFD-induced adipose tissue expansion was significantly attenuated with reduced vascular density and PCs detachment in the eWAT by Ana. To clarify the mechanism by which PCs attach even under HFD-fed condition by coexistence of SDF1 and PDGF-B, we isolated ECs, PCs, and macrophages (Mφ) from eWAT of mice. Expression of PDGFB was increased in Mφ and decreased in ECs by HFD, whereas these changes were attenuated and the gradient of PDGFB concentration between PCs and ECs was blunted in Ana-treated eWAT. These findings suggest that both PDGF-B and SDF1 coordinately regulate PCs detachments and adipose tissue expansion with obesity in mice.

Disclosure
T. Wada: Research Support; Self; Sanwa Kagaku Kenkyusho Co., Ltd.. E. Watanabe: None. Y. Kotera: None. Y. Onogi: None. H. Tsuneki: None. T. Sasaoka: None.
Interaction of Endothelial Cell Mineralocorticoid Receptor, Noncoding RNA, and Exosomes in Diet-Induced Vascular Stiffness and Insulin Resistance
Authors: JIA GUANGHONG , AROOR ANNAYYA R. , HABIBI JAVAD , WHALEY-CONNELL ADAM , SOWERS JAMES 
Published: June 2018

Insulin-mediated capillary recruitment is important for increasing the vascular surface area for both insulin and glucose delivery and subsequent muscle glucose-uptake. Our recent data suggest Western diet (WD) induces vascular stiffness and impairs insulin-mediated capillary recruitment by activation of cell-specific endothelial cell (EC) mineralocorticoid receptor (MR). Here, we further hypothesized interaction of ECMR, noncoding RNA, and exosome contributes to WD-induced impairment of vascular stiffness and insulin sensitivity. Six week-old ECMR KO and littermate wild type control male mice were fed a mouse chow or WD containing excess fat (46%) and fructose (17.5%) for 16 weeks. Insulin sensitivity and vascular activity were determined by intraperitoneal glucose tolerance test and wire myograph system. ECMR KO mice prevented WD-impaired systemic insulin sensitivity and improved the aortic vasodilation responses to acetylcholine (10-9-10-4 mol/L) and insulin (0.1- 300 ng/ml), which were impaired by WD. The impairment of insulin sensitivity and vascular function was associated with abnormal expression of noncoding RNA including microRNA (miR)-7, -23, -34, -92, -429, -449, -466 as well as long noncoding RNA-kis2, -Mirt1, -Nron, -Cox2 examined by RNA sequencing, which are associated endothelium inflammation and vascular remodeling. In vitro cultured ECs, aldosterone (10-7 mol) also increased EC exosome release and miR-103 expression. Furthermore, application of a miR103 inhibitor to ECs in vitro attenuated aldosterone-induced downregulation of serine (Ser) 1177/activation of endothelial nitric oxide synthase and upregulation of intercellular adhesion molecule 1. These findings suggest that increased ECMR signaling and associated abnormal expression and release of noncoding RNA and exosome play a key role in obesity-induced vascular stiffness and insulin resistance.

Disclosure
G. Jia: None. A.R. Aroor: None. J. Habibi: None. A. Whaley-Connell: None. J. Sowers: None.
Evidence of CB1-Receptor Antagonism in Fat-Fed Dogs Promoting Beiging of Adipose Tissue Mainly via Activation of Sarco/Endoplasmic Reticulum Ca2+ ATPase 2b and Ryanodine Receptor 2
Authors: PASZKIEWICZ REBECCA L. , BERGMAN RICHARD N. , RICHEY JOYCE M. , IYER MALINI S. , WOOLCOTT ORISON O. , KOLKA CATHRYN M. , CLEGG DEBORAH , KABIR MORVARID 
Published: June 2018

We have previously demonstrated that 16 weeks following exposure to a high fat diet (HFD), dogs treated with the CB1-receptor antagonist (Rimonabant (RIM)) upregulated β3-adrenergic receptors, PGC1α, Prdm16, and slight increase of UCP1 in the subcutaneous (SC) and visceral (VIS) depots when compared to control treated dogs. These data suggest an improvement in metabolic flux of adipose tissue consistent with beiging. The mechanism(s) by which RIM promotes adipose tissue beiging is unknown. We hypothesize that the beiging effect might be induced via UCP1-independent thermogenic mechanisms through upregulation of sarco/endoplasmic reticulum Ca2+-ATPase 2b (SERCA2b) and ryanodine receptor 2 (RyR2). To explore the mechanism by which RIM enhances adipose tissue function, dogs were fed a HFD (52% fat) for 6 weeks followed by a continued 16 weeks of fat feeding with either HFD + placebo (PL) (n=9) or HFD + RIM (1.25 mg/kg per day; n=11). Biopsies from SC and VIS depots were obtained for gene expression: before HFD (pre-fat) and 16 weeks of HFD +/- RIM. RIM increased SERCA2b expression in the SC depot by 8-fold (P&lt;0.05) and in the VIS depot by 2-fold (P&lt;0.05) compared to the PL+HFD groups. Also, RIM increased RyR2 3-fold in the SC (P&lt;0.005) and 4-fold in the VIS depots (P&lt;0.05). Our data suggest that a potential mechanism by which CB1-receptor antagonism increases beiging of adipose tissues may be through upregulation of the key genes involved in an UCP1-independent pathway. Importantly, for the first time we are beginning to identify mechanisms by which CB1-receptor antagonism enhances adipose tissue function and improves energy homeostasis in the presence of a HFD.

Disclosure
R.L. Paszkiewicz: None. R.N. Bergman: Advisory Panel; Self; Ingredion, Inc.. Research Support; Self; AstraZeneca. Consultant; Self; GI Dynamics Inc.. Research Support; Self; National Institute of Diabetes and Digestive and Kidney Diseases. Consultant; Self; Novo Nordisk A/S. J.M. Richey: None. M.S. Iyer: None. O.O. Woolcott: None. C.M. Kolka: Research Support; Self; AstraZeneca. D. Clegg: None. M. Kabir: None.
Gender-Dependent EPO Action in Adipose Tissue
Authors: LEE JEEYOUNG , NOGUCHI CONSTANCE T. 
Published: June 2018

Erythropoietin (EPO) is a glycoprotein that stimulates the production of red blood cells. Recent studies have shown the nonerythroid effect of EPO involved in metabolic regulation. EPO treatment attenuated body weight gain in male mice on normal chow, high fat diet induced obese mice and ob/ob mice. EPO regulation of body weight is concomitant with reduction in fat mass. EPO improved glycemic control in both male and female obese mouse models. However, the metabolic response to EPO to reduce weight gain primarily via reduced fat mass appears to be gender specific. Although EPO reduced weight gain in 6 month old female mice on high fat diet, regulation of body weight was not observed in young female mice on high fat diet. In contrast, young ovariectomized female mice on high fat diet showed EPO reduction in weight gain. This sex-specific reduction in weight gain is associated with female hormones as the decreased weight gain with EPO treatment in ovariectomized female mice on high fat diet is abrogated with estrogen supplementation. To determine specific in vivo consequences of potential gender differential response of white adipose tissue with EPO treatment, we examined signal transduction in diet induced obese mice following acute EPO treatment. Analyzing white adipose tissue harvested at 0, 30 min and 60 min following single dose administration of EPO (3000 U/kg), we found that EPO activated Erk signaling in white adipose tissue of male mice. In contrast, there was no evidence of acute Erk response in white adipose tissue of female mice following EPO treatment. Erk is known for its involvement in adipocyte differentiation, adiposity, and high-fat diet induced obesity. These data suggest that estrogen is a key modifier in the metabolic response to EPO, in part via EPO regulation of Erk signaling with gender specificity. Together, these results elucidate a potential mechanism for cross talk between EPO and estrogen, and provide a novel insight about gender specific EPO action to control adipose tissue development.

Disclosure
J. Lee: None. C.T. Noguchi: None.