bluuebunny/crossref_metadata_2025_split huggingface dataset:
1173-P: Sex Differences in NASH Pathways Informed by Multiomics
Authors: BLENCOWE MONTGOMERY , NORHEIM FRODE , SALEEM ZARA , HSU NEIL S. , HUI SIMON , PAN CALVIN , KRISHNAN KARTHICKEYAN CHELLA , EDILLOR CHANTLE , YANG XIA , LUSIS ALDONS 
Published: June 2021

Nonalcoholic fatty liver disease (NAFLD) has presented itself as a significant health burden, with increasing prevalence globally and evidence of a sexually dimorphic disease course. Particularly, it is of critical importance to understand the latter stage of disease, nonalcoholic steatohepatitis (NASH) with fibrosis, which is more prevalent and severe in males, in order to provide better prevention, treatment, and biomarker options to reduce the health burden. Our mechanistic understanding of NASH progression is far from elucidated, and thus, we present a multi-omics pathway and network analysis utilizing genomic, transcriptomic, and lipidomic data from the Hybrid Mouse Diversity Panel (HMDP) to uncover key biological processes and key driver genes causal to liver fibrosis and to dissect sex differences. Analyzing these multi-omics datasets by sex, we present mechanistic differences important to fibrosis development between males and females. We find that males present with greater enrichment of immune processes as well as abnormalities in protein and lipid metabolism, whereas females present with significant abnormalities linked with carbohydrate metabolism processes, suggesting differential causal mechanisms of fibrosis progression between sexes. Comparing males and females, we also find consistent processes such as extracellular matrix development, clearly vital for fibrosis to develop. Using gene regulatory network analysis of the liver tissue, we further identified key network genes driving the sex-specific processes as well as those consistent between sexes that are important for liver fibrosis. Our multi-omics integrative studies offer comprehensive understanding of NASH in a sex-specific manner.

Disclosure
M. Blencowe: None. A. Lusis: None. F. Norheim: None. Z. Saleem: None. N. S. Hsu: None. S. Hui: None. C. Pan: None. K. Chella krishnan: None. C. Edillor: None. X. Yang: None.

Funding
National Institutes of Health (R01DK117850)
1177-P: The Reduction in Hepatic Steatosis after Cushing’s Syndrome Treatment Is a Marker of Improving Glycemia
Authors: PIERCE ANNE , SATER ZAHRAA A. , CYPESS AARON M. 
Published: June 2021

Cushing Syndrome (CS) is characterized by cortisol excess that leads to impaired glucose tolerance and obesity. CT imaging shows 20% of CS patients have nonalcoholic steatohepatitis (NASH). While the gold standard diagnostic test for NASH is liver biopsy, it has too much risk for CS patients. This study evaluated the ability of noninvasive, magnetic resonance spectroscopy (MRS) to measure liver fat percentage in CS patients and followed temporal changes in metabolic parameters before and after successful treatment. The primary outcome was protein density fat fraction (PDFF) by MRS at 3 timepoints: baseline, 6 months, and 12 months after treatment, which consisted of surgery as indicated based on ACTH source. The diagnosis was confirmed if there was histology consistent with corticotrope adenoma, adrenal tumor, or an ectopic source. We prospectively studied 41 consecutive patients: 85% female; 44±1.8 y; 32.6±1.5 kg/m²; urine cortisol excretion 2242.7±1806.3 [nl 3.5-45.0 mcg/24h]). Each underwent MRS at 3T where NASH was defined as &gt;5% PDFF. A1c and BMI were measured. Changes over time were evaluated using Wilcoxon signed-rank test, and correlation among variables was done with Spearman’s rank test. At baseline, mean PDFF was 10.4±1.7 and correlated positively with BMI (r=0.57, P&lt;0.001). NASH was present in 32% of patients. PDFF reductions were similar after 6 and 12 months of treatment (-52%, P=0.001 and -50%, P=0.02); rates of NASH declined to 13% and 11%; BMI decreased (-9%; P=0.002 and -12%, P&lt;0.001). A1c was lower at 12 months (6.2±1.1 to 5.6±0.5, P&lt;0.001). The reduction in liver fat correlated with changes in visceral fat mass (r=0.70, P&lt;0.001).
In summary, MRS-PDFF was effective at diagnosing NASH in CS. Physiologically, definitive treatment of CS reduced liver fat by 6 months after normalization of cortisol and also preceded the improvement in A1c. These results suggest that liver insulin resistance due to fat accumulation has an important role in diabetes pathophysiology in CS.

Disclosure
A. Pierce: None. Z. A. Sater: None. A. M. Cypess: None.
1189-P: Ankyrin-B Modulates the Integrity and Bioenergetic Capacity of Skeletal Muscle Fibers
Authors: VOOS KAYLEIGH , TZENG JOYCE , LORENZO DAMARIS 
Published: June 2021

Ankyrin-B (AnkB) is a cytoskeletal protein that positions and stabilizes specialized membrane-associated protein complexes. AnkB also associates with organelles and facilitates their dynamics across cell types. AnkB variants have been linked to higher risk for obesity and type 2 diabetes (T2D) in humans and mice. In skeletal muscle (SKM), AnkB localizes to costameres and acts as a scaffold to β-dystroglycan/dystrophin complexes. This protein complex has been demonstrated to be essential for SKM integrity and exercise injury protection. However, besides this structural role, our findings suggest that AnkB associates with mitochondria in SKM and likely plays important roles in SKM bioenergetics. We found that non-obese young mice lacking AnkB solely in SKM (SKM-AnkB KO) develop early signs of myofibrillar disorganization and fibrosis and display reduced physical activity without changes in body weight, fasting glucose levels, or peripheral insulin response. In contrast, SKM-AnkB KO mice are more susceptible to the metabolic effects of a HFD, manifested as lower insulin sensitivity and further reductions in physical activity. Our findings suggest that loss of myofibril organization synergizes with changes in mitochondrial function to worsen SKM bioenergetics and insulin action. These physiological alterations are in agreement with our RNA-seq and proteomics studies, which implicate AnkB in the regulation of SKM oxidative capacity, protein translation, and cellular responses to stress and nutrient availability. We will discuss our ongoing efforts to uncover the mechanistic basis of these metabolic impairments cause by AnkB deficits.

Disclosure
K. Voos: None. J. Tzeng: None. D. Lorenzo: None.

Funding
American Diabetes Association (1-19-JDF-081 to D.L.)
119-OR: Ankyrin-B Modulates Lipid Metabolism in Brown Adipocytes
Authors: AGUILLARD ASHLEY , TZENG JOYCE , LORENZO DAMARIS 
Published: June 2021

Human variants in the membrane adaptor ankyrin-B (AnkB) present at frequencies from 0.05 to 7% across different ethnicities have been linked to obesity and type 2 diabetes (T2D). We reported that knock-in mice bearing T2D-linked human AnkB variants develop age- or diet-dependent obesity and insulin resistance (IR). We also found that conditional loss of AnkB in both white (WAT) and brown (BAT) adipose tissue (AT) in mice (AT-AnkB KO) results in age-dependent obesity with WAT hypertrophy, lipotoxicity, systemic inflammation, and IR. We delineated that defects in the internalization of the glucose transporter GLUT4 in WAT, which resulted from deficits in AnkB, caused cell-autonomous WAT hyperplasia and ectopic lipid deposition in peripheral tissues. AT-AnkB KO mice also exhibit lower energy expenditure (EE) before the onset of obesity, which is concomitant with reduced BAT mitochondrial content and brown adipocyte whitening. However, whether primary BAT dysfunction caused by AnkB deficits contribute to metabolic syndrome in these mice and the roles of AnkB in BAT is unknown. Here, we present evidence of a cell-autonomous role of AnkB in lipid metabolism in brown adipocytes. Histological and electron microscopy analyses suggest that conditional AnkB loss in BAT of non-obese mice (BAT-AnkB KO) causes lipid accumulation in BAT characterized by an increase in surface area covered by lipid droplets, a shift towards larger lipid droplets, and a corresponding decrease in mitochondrial content. Differentiated brown adipocytes from mice with total AnkB deficiency (AnkB KO) or from BAT-AnkB KO mice phenocopy the in vivo changes in lipid and mitochondria content, which corresponds with changes in associated gene expression. We will discuss our latest results combining physiology, transcriptomic, proteomic, and cellular analysis that shed light into the pathways modulated by AnkB in BAT and their contribution to systemic metabolic regulation.

Disclosure
A. Aguillard: None. J. Tzeng: None. D. Lorenzo: None.

Funding
National Institutes of Health (3P30DK072476-13S1)
1196-P: Perineuronal Nets in MeA and Arc Are Influenced by Gonadectomy
Authors: ZHANG NAN , XU YONG 
Published: June 2021

Background: Recent studies reveal perineuronal nets (PNNs), extracellular matrix specializations that restrict the plasticity of neurons, are influenced by pubertal status. Here we show that PNN are changed by gonadectomy (GDX) both in male and female mice in medial amygdala (MeA) and arcuate nucleus (Arc).
Method: 49 wild type mice are divided into 16 groups by sex, gonadectomy, diet, nutritional status (fed or fasted groups). Fat and lean body mass were assessed by QMR at baseline and before perfusion. Body weight, food intake, and blood glucose were measured weekly for five weeks. Images were captured in MeA, Arc, paraventricular hypothalamus (PVH), anterior hypothalamus (AH), and an intense WFA positive radial area near PVH, which we named ‘periPVH’ after labelling of Wisteria floribunda agglutinin (WFA).
Results: In MeA region, female mice have significantly lower total fluorescence intensity (TFI) (Estimate, 95% Confidence Interval: –448887.02, –578807.10 – –318966.94; p &lt;0.0001) and total number of PNNs (–319.67, –379.34258 – –259.99075: p&lt;0.0001) than male mice. Notably, in female, OVX mice have lower mean fluorescence intensity (MFI) (Mean ± SD: 14.5 ± 3.3 vs. 19.6 ± 4.1; p = 0.003) and TFI (64907.8 (58406.6–96971.3) vs. 123613.9 (83545.9–149131.5); p = 0.015) than sham mice, while the total number of PNNs (Mean ± SD: 63 ± 23 vs. 55 ± 3; p = 0.391) shows no significant difference. Male castration mice have lower total number of PNNs (382 ± 141 vs. 55 ± 22; p &lt;0.0001) and TFI (439343.5 (366431.4–787461.9) vs. 168416.3 (141273.0–258020.8); p &lt;0.0001) than sham mice, while the MFI shows no significant difference. In Arc, mice experiencing GDX have significantly lower MFI (–3.63, –5.61303 – –1.63967; p &lt;0.001), TFI (–28525.8, –45357.46 – –11694.16; P&lt;0.01) or total number of PNNs (–15.3, –29.24 – –1.41; p =0.036) than mice not. PVH, AH, and periPVH don’t show associations with sex, GDX, diet or nutritional status in our observation.
Summary: Our novel discovery indicates the potential role PNN might play in sex hormone regulation in brain.

Disclosure
N. Zhang: None. Y. Xu: None.

Funding
National Institutes of Health (R01DK117281)
1202-P: Hepatic NgBR Is a Potential Resilience Factor for Preventing Obesity-Caused Insulin Resistance
Authors: HU WENQUAN , WANG XIANG , MIAO QING 
Published: June 2021

Adiponectin is an adipocyte-secreted protein hormone and prevents the development of insulin resistance. The adiponectin-mediated activation of AMP-activated protein kinase (AMPK) pathway is essential for decreasing the expression of hepatic gluconeogenic enzymes and enhancing the hepatic effects of insulin. However, the resilience factors required to maintain adiponectin-mediated AMPK activation signaling in the liver remain unclear. Recent studies from our laboratory suggest that NgBR plays a previously unrecognized role in preserving AMPK activation. The loss of NgBR in the obesity liver impairs adiponectin-dependent gluconeogenesis and insulin sensitivity. NgBR hepatocyte-specific knockout (hepKO) mice resemble many features of prediabetes, such as moderately increased fasting blood glucose levels and insulin resistance, as well as facilitating the high-fat diet (HFD)-induced onset of T2D. NgBR was identified as a cell surface receptor for soluble Nogo-B (sNogo-B). Also, NgBR binds the farnesylated form of liver kinase B1 (LKB1-farn), a key regulator of AMPK. NgBR depletion in hepatocytes abolishes the localization of LKB1 in the plasma membrane and impairs AMPK activation, the critical signal for adiponectin-mediated insulin sensitivity. NgBR expression is decreased in the liver of diabetic mice. Similar changes also occur in diabetes patients. The NgBR transcript level is reduced in T2D obese patients’ liver compared to non-T2D obese and lean subjects. Additional in vitro studies show that NgBR deficiency in hepatocyte prevents AMPK activation induced by AdipoRon (an agonist of adiponectin receptor) and increases the gluconeogenesis. Our data suggest that NgBR is a potential resilience factor required for preserving adiponectin-mediated insulin sensitivity and preventing the onset of obesity-caused T2D, and disruption of the NgBR-dependent regulation system leads to impairing gluconeogenesis regulation and insulin sensitivity in the liver.

Disclosure
W. Hu: None. X. Wang: None. Q. Miao: None.

Funding
National Institutes of Health (R01DK112971)
1206-P: Effects of Short-Term Administration of Delta-9-tetrahydrocannabinol on Metabolic Parameters of C57bl/6 Mice
Authors: KURJAN EMILY , OLSON LAWRENCE K. , ALSHAARAWY OMAYMA 
Published: June 2021

Cancer and HIV-related cachexias are among the qualifying conditions for medical cannabis use. Although effective to alleviate nausea and vomiting, clinical trials demonstrated little to no impact of cannabinoids on weight gain in patients. Consistent with this, cannabis users have a lower prevalence of obesity in epidemiological studies. Since cannabinoid-1 receptors (CB1R) regulate food intake and adipose tissue thermogenesis, we studied the metabolic effects of a low dose of delta-9-tetrahydrocannabinol (THC), a CB1R agonist and the main bioactive chemical in cannabis, in 8-week old male mice maintained on a 10%-fat-calorie diet. Mice received daily intraperitoneal injections of THC (3 mg/Kg, n = 15) or vehicle (n = 12) just before the dark cycle for 7 days. THC-treated mice gained an average of 1.6 grams compared to 2.6 grams gained by the vehicle-treated mice (p = 0.03). Despite the attenuated weight gain, automated home cage phenotyping revealed an increase in food intake in the 90 minutes after injections in the THC-treated mice (mean = 0.78 vs. 0.53 grams; p = 0.001). No differences, however, were detected in overall light (p = 0.39) or dark-cycle food intake (p = 0.07). THC-treated mice also showed reduced locomotor activity in the dark cycle compared to vehicle-treated mice (p &lt;0.001). There were no robust differences in indirect calorimetry parameters between groups in the dark cycle, whereas the respiratory exchange rates were lower among THC-treated mice during the light cycle (p = 0.01). Gene expression studies revealed reduced CB1R mRNA levels in brown adipose (mean = 0.61; p = 0.02) and liver (mean = 0.30; p = 0.04) of THC-treated mice.
In summary, administration of a low dose of THC was associated with attenuated weight gain in male C57BL/6 mice, possibly through downregulation of CB1R in metabolically active tissues. Future studies will elucidate the effects of chronic THC administration on CB1R expression and thermogenesis under high fat feeding and obesity.

Disclosure
E. Kurjan: n/a. L. K. Olson: None. O. Alshaarawy: None.

Funding
National Institutes of Health (R00AT009156)
1227-P: Association between Fat-to-Muscle Ratio and Insulin Resistance in Adults with Impaired Fasting Glucose
Authors: LAI YUN-JU 
Published: June 2021

Background: Body composition measurements are useful for assessing cardiometabolic risk. But the associations of the fat-to-muscle ratio with insulin resistance was not well investigated.
Methods: Adults with impaired fasting glucose were recruited from a community hospital in Taiwan during 2017. Demographics, lifestyle features, and medical history were gathered by well-trained interviewers. All participants underwent comprehensive physical examinations, including a biochemical assay of venous blood specimens and urine samples after an 8-hour overnight fast. Fat-to-muscle ratio was measured using whole-body dual-energy X-ray absorptiometry and calculated as follows: total fat mass (g) divided by total lean mass (g). The homeostasis model assessment for IR (HOMA-IR) was used to evaluate insulin resistance and was calculated as follows: [fasting plasma glucose level (mg/dL) × fasting plasma insulin level (uIU/mL)]/405. Multivariable logistic regression analysis was employed to explore the relation between fat-to-muscle ratio and insulin resistance.
Results: Among 234 participants, the proportion of women was 50.1%, and the mean age was 64.2 ± 10.0 years. The fat-to-muscle ratio significantly differed between men and women (0.38 ± 0.12 vs. 0.62 ± 0.16, respectively, P &lt; 0.001). Insulin resistance is significantly associated with fat-to-muscle ratio increased (P&lt;0.001). The multivariable-adjusted odds ratio of fat-to-muscle ratio for insulin resistance (HOMA-IR ≥ 3) were 65.38 [95% confidence interval (CI): 4.03- &gt;999.999] in men, and 12.73 (95% CI: 2.64- 61.41) in women.
Conclusions: The current study revealed that a high fat-to-muscle ratio was significantly associated with increased risk of insulin resistance in adults with impaired fasting glucose.

Disclosure
Y. Lai: None.
123-OR: Deletion of Both Pcsk1 And Pcsk2 in Beta Cells: Lack of Mature Insulin Drives Beta-Cell Expansion and Dysfunction, but Not Severe Diabetes in Mice
Authors: TAYLOR AUSTIN , CHEN YI-CHUN , PASULA DANIEL , LUCIANI DAN S. , VERCHERE BRUCE 
Published: June 2021

Proinsulin (PI) is processed to mature insulin by prohormone convertases 1/3 (Pcsk1) and 2 (Pcsk2). In individuals with T1D and T2D, there is an increase in the circulating PI:insulin ratio. We hypothesized that β-cell Pcsk1 and Pcsk2 deletion would result in β-cell dysfunction, loss of insulin production, and marked hyperglycemia.
Single Pcsk1flox/flox Ins1cre/+ (Pcsk1βKO), Pcsk2flox/flox Ins1cre/+ (Pcsk2βKO), and double Pcsk1flox/flox Pcsk2flox/flox Ins1cre/+ (DPCβKO) β-cell prohormone convertase knockout mice were generated (with respective Pcsk1+/+ Pcsk2+/+ Ins1cre/+ controls) and monitored until 30 weeks old. Pcsk2βKO mice displayed no glycemic phenotype, while Pcsk1βKO mice displayed mild hyperglycemia and impaired glucose tolerance by 26 weeks of age. DPCβKO mice displayed mild hyperglycemia (10.1±1.6 vs. 12.3±2.1 mM; p=0.006) and marked glucose intolerance by 10 weeks of age (AUC: 418±135 vs. 1485±360; p&lt;0.0001). No animals displayed obesity relative to controls. Insulin+ pancreas area was doubled in DPCβKO male mice (p=0.0004) and DPCβKO islets showed early glucose-induced Ca2+ influx and loss of oscillatory Ca2+ flux. Pcsk2βKO islets had unaltered mature insulin, while Pcsk1βKO islets had reduced mature insulin. DPCβKO islets contained no detectable mature insulin by western blot. In contrast, Pcsk2βKO islets had markedly reduced mature islet amyloid polypeptide (IAPP), while Pcsk1βKO islets contained normal mature IAPP levels.
Our results suggest that complete loss of β-cell prohormone processing, including PI processing, can drive β-cell dysfunction and expansion, but only mild diabetes and glucose intolerance. Although Pcsk2 deletion in β cells does not impact glycemia or PI processing, it is required for normal proIAPP processing. In total, our data support the idea that altered β-cell prohormone processing does not impact obesity but can contribute to the pathogenesis of diabetes via β-cell dysfunction.

Disclosure
A. Taylor: None. Y. Chen: None. D. Pasula: None. D. S. Luciani: None. B. Verchere: Advisory Panel; Self; Integrated Nanotherapeutics Inc., Sirona Biochem, Stock/Shareholder; Self; Integrated Nanotherapeutics Inc.

Funding
Canadian Institutes of Health Research
1237-P: Generation of Targeted Nanoparticles for ß-Cell-Selective Delivery of Antioxidant Drugs
Authors: NOVAK ALISSA , CHAKRAVARTHULA TANMAYE NALLAN , CROWDER JUSTIN , ALVES NATHAN J. , BROICHHAGEN JOHANNES , HODSON DAVID , BEDWELL AMANDA , LINNEMANN AMELIA K. 
Published: June 2021

β-cells are highly metabolic and vulnerable to a variety of exogenous stressors, many of which elicit the generation of reactive oxygen species (ROS). Endogenous adaptive responses can restore homeostasis after excessive ROS elevation. If the endogenous adaptive response fails, ROS accumulation can cause damage to DNA, proteins, and organelles, ultimately leading to apoptosis. A wealth of evidence supports that ROS accumulation contributes to β-cell dysfunction and destruction during diabetes development. These findings suggest that stimulating the antioxidant response could prevent or treat diabetes. However, limitations exist in that persistent systemic activation of the antioxidant response can perturb the normal second messenger function of ROS, and that exogenous antioxidants have poor bioavailability and lack specificity. To overcome these obstacles, we developed β-cell targeted nanoparticles to selectively deliver antioxidants to β-cells. We conjugated modified Exendin-4 (Ex4) to an amphiphilic polymer, allowing us to generate lipid nanoparticles to deliver small hydrophobic compounds to GLP-1R expressing cells. We assessed β-cell targeting of our nanoparticles using isolated islets from mice expressing tdTomato selectively in the β-cell. Isolated islets were treated with nanoparticles containing a lipid intercalating dye, DiO, and we observed selective DiO accumulation in tdTomato positive cells. We also performed in vivo imaging of live mice injected with targeted or non-targeted nanoparticles containing a near-IR lipid intercalating dye. These experiments demonstrated selective retention of targeted nanoparticles both systemically and in the pancreata of animals 1 hour after tail vein injection. Collectively, we provide evidence that Ex4 coated micelles are selective to β-cells. Future experiments will explore this targeted approach to deliver drugs to stimulate the antioxidant response and reduce β-cell apoptosis under diabetogenic conditions.

Disclosure
A. Novak: None. T. Nallan chakravarthula: None. J. Crowder: None. N. J. Alves: None. J. Broichhagen: None. D. Hodson: None. A. Bedwell: None. A. K. Linnemann: None.