Hepatic Fat Research Articles

Bean Leaves Ameliorate Lipotoxicity in Fatty Liver Disease.

Bioactive compounds in plant-based food have protective effects against metabolic alterations, including non-alcoholic fatty liver disease (NAFLD). Bean leaves are widely cultivated in the world and are a source of dietary fiber and polyphenols. High fat/high fructose diet animal models promote deleterious effects in adipose and non-adipose tissues (lipotoxicity), leading to obesity and its comorbidities. Short-term supplementation of bean leaves exhibited anti-diabetic, anti-hyperlipidemic, and anti-obesity effects in high-fat/high-fructose diet animal models. This study aimed to evaluate the effect of bean leaves supplementation in the prevention of lipotoxicity in NAFLD and contribute to elucidating the possible mechanism involved for a longer period of time. During thirteen weeks, male Wistar rats (n = 9/group) were fed with: (1) S: Rodent Laboratory Chow 5001® (RLC); (2) SBL: 90% RLC+ 10% dry bean leaves; (3) H: high-fat/high-fructose diet; (4) HBL: H+ 10% of dry bean leaves. Overall, a HBL diet enhanced impaired glucose tolerance and ameliorated obesity, risk factors in NAFLD development. Additionally, bean leaves exerted antioxidant (↑serum GSH) and anti-inflammatory (↓mRNA TNFα in the liver) effects, prevented hepatic fat accumulation by enhanced ↑mRNA PPARα (β oxidation), and enhanced lipid peroxidation (↓liver MDA). These findings suggest that bean leaves ameliorated hepatic lipotoxicity derived from the consumption of a deleterious diet.

Loss of epidermal glucocorticoid receptor protects against whole body metabolic dysfunction upon chronic corticosterone treatment

ObjectiveGlucocorticoid (GC) excess contributes to the development of metabolic syndrome, defined by visceral obesity, abnormal glucose tolerance, and dyslipidemia. While it is accepted that loss of metabolic control is causative of cutaneous diseases, the systemic effects of epidermal dysfunction have received limited attention. Importantly, independent of GC blood levels, skin synthesis of these hormones can provide tissue-specific variations that may affect global homeostasis. We aimed to assess whether the epidermal-specific loss of the GC receptor (GR) had an impact on the dermal white adipose tissue (dWAT), a specialized fat depot functionally different from other fat pads, as well as on whole body homeostasis. MethodsGR epidermal KO (GREKO) and control female mice were treated with oral corticosterone (CORT) for 4 weeks, a protocol inducing metabolic dysfunction. Metabolic parameters including body weight, visceral and hepatic fat accumulation, blood glucose and insulin levels, glucose tolerance tests upon fasting, and triglycerides levels, were determined. Systemic alterations of soluble factors with known roles in immunity and inflammation were also assessed by a multiplex antibody array system containing selected cytokines, chemokines, and growth factors. The levels of cutaneous GCs and the profile of skin-secreted factors were determined in tissue explants by ELISA and the multiplex array system. Morphometric studies quantitated changes in dWAT thickness and adipocyte size in both genotypes, basally and at the end of CORT treatment. The expression of adipocyte markers was assessed in purified dermal adipocytes in vehicle and CORT-treated GREKOvs controls. ResultsDespite similar circulating levels of GCs, GREKO mice were highly resistant to CORT-induced systemic metabolic anomalies including body weight gain, visceral and hepatic fat, hyperglycemia, insulinemia, and elevated levels of plasma triglycerides, leptin, FGF-21, PAI-1, and CCL11. GREKO mice featured constitutively enhanced levels of cutaneous GCs relative to controls at least partially due to keratinocyte-specific increased expression of the critical steroidogenic enzyme Cyp11b1. Also, the higher ratio of skin-secreted protective vs inflammatory adipokines in GREKOvs controls, correlated with higher capacity of adipogenic conversion in experiments using conditioned media from tissue explants. Following CORT treatment, relative to controls, GREKO mice featured reduced dWAT hyperplasia and adipocyte hypertrophy, with increased Adipoq and decreased Lipocalin 2 expression in purified dermal adipocytes. ConclusionsOverall data suggest that epidermal GR loss results in paracrine actions on dermal adipocytes as well as endocrine actions on key metabolic tissues that significantly improve the whole body metabolism in a mouse model of metabolic dysfunction.

Assessment of Segmental Hepatic Fat Distribution using Magnetic Resonance Proton Density Fat Fraction MR-PDFF in Non-alcoholic Fatty Liver Disease (NAFLD)

Abstract: Background: Non-Alcoholic Fatty Liver Disease (NAFLD) is a significant healthcare challenge. MR proton density fat fraction (MR-PDFF) is a quantitative imaging parameter that allows a precise estimation of hepatic steatosis. Determination of segmental and lobar fat distribution is also important since underestimation or overestimation may lead to hurdles in patient management and may also alter outcomes during liver donor assessment for living donor liver transplant. Objective: To determine the heterogeneity of hepatic fat distribution across different liver segments and both lobes in patients with non-alcoholic fatty liver disease (NAFLD). Materials and Methods: This cross-sectional descriptive study included 35 patients of NAFLD. MR-PDFF sequence was performed, two regions of interest (ROI) were drawn at the periphery of each hepatic segment and their mean was taken. We calculated mean values, ranges, and standard deviations for individual segments, both lobes and the entire liver. Pearson’s correlation was used to assess the relation between MR-PDFF and MR-PDFF variability. Paired sample t-test was utilized to compare the means of the right and the left lobe of the liver. Results: The fat fraction in segment I was the lowest and in segment VII the highest. The right and left lobes showed a significant difference in fat fraction with values of 14% and 11.4% respectively (paired sample t-test, p<0.005). The left lobe showed a greater MR-PDFF variability than the right lobe (1.9 vs 1.6%). Conclusion: In patients with NAFLD, segments VII and VIII show the greatest while segments I and IV show the least fat infiltration. Hepatic fat preferentially gets deposited more in the right lobe of the liver.

Validation of MRI Quantification of Liver Fat, Keeping CT Liver Attenuation Index (LAI) as Gold Standard

Abstract: Background: Non – Alcoholic Fatty Liver Disease (NAFLD) is emerging as a considerable health problem in patients visiting gastroenterology clinics. It is of crucial importance to evaluate the extent of hepatic steatosis in potential candidates for living donor liver transplantation (LDLT) to ensure donor safety as well as optimum graft regeneration. Objective: To validate the MRI quantification of liver fat keeping CT liver attenuation index as gold standard. Materials and Methods: This cross-sectional study was carried out in the department of Diagnostic Radiology, Pakistan Kidney and Liver Institute and Research Centre from 10th October, 2022 to 10th December, 2022. We determined the sample size using WHO sample size calculator. The MR fat fraction sequence was acquired as a part of the obligatory MRCP in 70 potential liver donors who undertook CT abdomen. Liver Attenuation Index (LAI) and MR fat fraction were determined separately by two radiologists who were blinded to each other. LAI was calculated as: Mean liver attenuation – mean splenic attenuation. MRI fat fraction from seven areas of liver were taken and their mean calculated to determine the percentage of liver fat. SPSS version 20 was employed for statistical analysis and Pearson’s Correlation was applied. Results: Among the 70 donors 42 were males and 28 were females (M: F= 1.5: 1). The hepatic fat fraction values on MR were correlated with the liver attenuation index on CT using a two – tailed Pearson correlation test. The results showed a very strong negative correlation between the two; the lower the LAI, the higher the MR fat fraction (Pearson correlation coefficient r = -0.932, p<0.05). Conclusion: Strong correlation was found between MRI estimation of liver fat and CT LAI fat estimation. MRI is safer than CT as it does not involve ionizing radiation, is quicker to perform, and hence can be recommended as future method of choice.

Effect of laparoscopic sleeve gastrectomy on mobilization of site-specific body adipose depots: a prospective cohort study.

Effect of bariatric surgery on mobilization of site-specific body adipose depots is not well investigated. Herein, the authors conducted a prospective cohort study to assess whether bariatric surgery can differentially affect specific fat storage pools and to further investigate correlations between site-specific fat mobilization and clinical outcomes. In this single-centre prospective cohort study, 49 participants underwent laparoscopic sleeve gastrectomy (LSG) from 24 May 2022 to 20 October 2022 and underwent MRI to estimate subcutaneous fat area, visceral fat area (VFA), hepatic and pancreatic proton density fat fraction (PDFF) at baseline and 3 months after surgery. The protocol for this study was registered on clinicaltrials.gov. Among 49 patients who met all inclusion criteria, the median [interquartile range (IQR)] age was 31.0 (23.0-37.0) years, the median (IQR) BMI was 38.1 (33.7-42.2)kg/m 2 and 36.7% were male. Median (IQR) percentage hepatic PDFF loss was the greatest after bariatric surgery at 68.8% (47.3-79.7%), followed by percentage pancreatic PDFF loss at 51.2% (37.0-62.1%), percentage VFA loss at 36.0% (30.0-42.4%), and percentage subcutaneous fat area loss at 22.7% (17.2-32.4%) ( P <0.001). By calculating Pearson correlation coefficient and partial correlation coefficient, the positive correlations were confirmed between change in VFA and change in glycated haemoglobin ( r =0.394, P =0.028; partial r =0.428, P =0.042) and between change in hepatic PDFF and change in homoeostatic model assessment of insulin resistance ( r =0.385, P =0.025; partial r =0.403, P =0.046). LSG preferentially mobilized hepatic fat, followed by pancreatic fat and visceral adipose tissue, while subcutaneous adipose tissue was mobilized to the least extent. Reduction in visceral adipose tissue and hepatic fat is independently associated with the improvement of glucose metabolism after LSG.

Exploring the Impact of Nutrition on Non-Alcoholic Fatty Liver Disease Management: Unveiling the Roles of Various Foods, Food Components, and Compounds.

There is a need to introduce standardized treatment options for non-alcoholic fatty liver disease (NAFLD) due to its global prevalence and the complications of this disease. Many studies have revealed that food-derived substances may be beneficial in dealing with this disease. Therefore, this review aims to evaluate the recently published studies on the food-derived treatment options for NAFLD. A comprehensive search of the PubMed database using keywords such as "NAFLD", "nutrition", "food", "derived", "therapy", and "guidelines" yielded 219 relevant papers for our analysis, published from 2004 to 2023. The results show the significant benefits of food-derived treatment in NAFLD therapy, including improvements in liver histology, hepatic fat amounts, anthropometric measures, lipid profile, and other metabolic measures. The availability of the substances discussed makes them a significant adjuvant in the treatment of this disease. The usefulness of Viusid as additional therapy to diet and physical activity should be emphasized due to improvements in liver histology; however, many other substances lead to a decrease in liver fat amounts including, e.g., berberine or omega-3 fatty acids. In addition, the synbiotic Protexin seems to be useful in terms of NAFLD treatment, especially because it is effective in both obese and lean subjects. Based on the latest research results, we suggest revising the therapeutic recommendations for patients suffering from NAFLD.

1090-P: Association between Organ Fat and Type 2 Diabetes Risk Factors among High-Risk Latino Youth

Obesity is a well-established risk factor for type 2 diabetes (T2D) in youth. However, the contribution that organ fat plays to the pathophysiology of T2D among high-risk youth is sparsely documented. The purpose of this study was to determine the independent associations between hepatic and pancreatic fat fractions with insulin sensitivity and beta-cell function among Latino youth with obesity and prediabetes. This cross-sectional analysis involved 117 Latino adolescent boys (n=70) and girls (n=47), mean (±SD) age:14±1 years, fasting glucose: 102±8mg/dL, 2-hour glucose: 144±30mg/dL, and with obesity (BMI percentile: 98.52±1.13). Total body composition was assessed by DEXA, while hepatic and pancreatic fat contents were assessed using quantitative fat fraction magnetic resonance imaging at 3 Tesla. T2D risk factors included insulin sensitivity (Matsuda index) and beta-cell function (product of insulin sensitivity and insulinogenic index), estimated from a multi-sample 2-hour oral glucose tolerance test. Associations between organ fat and T2D risk factors were examined using Pearson correlation and multiple linear regression, adjusting for age, sex, and total body fat percentage (BF%). Insulin sensitivity significantly correlated with only BF% (r =-0.236, p=0.015), fat mass (r=-0.227, p=0.020), and hepatic fat fraction (r=-0.255, p=0.024), whereas beta-cell function significantly correlated with only BF% (r=-0.241, p=0.013). In multiple regression models, hepatic fat fraction (B= -0.045, p=0.041, Δr2=.034) but not pancreatic fat fraction (B= 0.058, p=0.179, Δr2= .015) was independently associated with insulin sensitivity, whereas neither measure of organ fat was associated with beta-cell function (each Δr2 &amp;lt;.01, each p&amp;gt;.38). Hepatic fat content was an independent predictor of insulin sensitivity among Latino youth with prediabetes. Further studies are needed to understand how adiposity contributes to beta-cell dysfunction among high-risk youth. Disclosure E.O.Owolabi: None. M.Olson: Advisory Panel; Rhythm Pharmaceuticals, Inc. H.H.Hu: None. A.Pena: None. K.A.Pituch: None. S.Bailey: None. W.C.Knowler: None. G.Q.Shaibi: None. Funding National Institute of Diabetes and Digestive and Kidney Diseases (R01DK107579, F31DK125037)

262-LB: Fasting Intact Insulin by Liquid Chromatography–Tandem Mass Spectrometry (LC-MS/MS) as a Predictor of Nonalcoholic Fatty Liver Disease in Pediatric Patients

Background: Recent research in adults has shown fasting intact insulin levels measured by mass spectrometry can predict nonalcoholic fatty liver disease (NAFLD) in patients without diabetes; however, there is limited research in children. This study aimed to evaluate fasting intact insulin by LC-MS/MS as a predictive biomarker of NAFLD in adolescents. Methods: This retrospective, cross-sectional study was based on data from four previously conducted studies. Existing clinical laboratory values included fasting intact insulin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), C-peptide, and the insulin resistance (IR) score. Hepatic fat was assessed by magnetic resonance imaging proton density fat fraction (MRI-PDFF) or MR spectroscopy (MRS). NAFLD was defined as intrahepatic fat of ≥5%. Results: A total of 73 children were included in the analysis (64% male; 13.9 ± 2.8 years of age; body mass index (BMI) 29.1 ± 8.5 kg/m2; 56% with NAFLD). Intact fasting insulin, C-peptide, AST, ALT, and IR scores were higher in children with NAFLD compared to non-NAFLD children (P&amp;lt;0.05). Fasting intact insulin measurements predicted NAFLD (area under the receiver operating characteristic curve [AUROC] of 0.83 [95%CI, 0.73-0.93]). When combined with ALT (intact insulin (μU/mL) × ALT (× U/L)), the combined biomarker slightly improved the prediction of NAFLD (AUROC 0.89 [0.81-0.96]), with positive and negative predictive values of 79% and 100%, respectively. Based on maximizing Youden's index, the optimal cutoff points to predict NAFLD were 13.0 μU/mL for intact insulin and 380 μU/mL × U/L for intact insulin × ALT. Conclusion: Our findings support that fasting intact insulin alone, or combined with ALT (intact insulin × ALT), show promise as predictive biomarkers for pediatric NAFLD with high positive and negative predictive values. Additional studies are needed to validate these findings in other pediatric populations. Disclosure H. Huneault: None. D. Ayinde: None. Z. He: None. C. C. Cohen: None. M. J. McPhaul: Employee; Quest Diagnostics. M. B. Vos: Consultant; Boehringer Ingelheim International GmbH, BMS, Intercept Pharmaceuticals, Inc., Novo Nordisk, ProSciento, Eli Lilly and Company. Research Support; Quest Diagnostics, TARGET PharmaSolutions, Inc. Funding National Institutes of Health (R01DK125701); Georgia Clinical Translational Science Alliance (UL1TR002378)

1558-P: The Mitochondrial Calcium Uniporter Regulates Hepatic Mitochondrial Oxidation and Intracellular Redox In Vivo

The mitochondrial calcium uniporter (MCU) facilitates mitochondrial calcium influx, and several enzymes that catalyze key steps of the TCA cycle require calcium for their activation. As such, it is generally accepted that rates of mitochondrial oxidation correlate with mitochondrial calcium activity. Decreased rates of hepatic mitochondrial oxidation are associated with metabolic dysfunction and ectopic lipid accumulation as seen with type 2 diabetes and nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis. Despite this, a role for MCU and mitochondrial calcium in regulating hepatic mitochondrial oxidation is still unclear. To address this question, we generated a liver-specific MCU knockout (MCU KO) mouse model which displayed reduced mitochondrial calcium ([Ca2+]mt; p&amp;lt;0.05) and increased cytosolic calcium ([Ca2+]cyt; p&amp;lt;0.01). Despite decreased [Ca2+]mt, we found that deletion of hepatic MCU increased rates of isocitrate dehydrogenase flux (VIDH; p&amp;lt;0.01), α-ketoglutarate dehydrogenase flux (VOGDH; p&amp;lt;0.01), and succinate dehydrogenase flux (VSDH; p&amp;lt;0.05) in vivo, using a novel [13C5]glutamine tracer infusion technique (Q-Flux). Further, we show that rates of [14C16]palmitate oxidation (p&amp;lt;0.01) and intrahepatic lipolysis (p&amp;lt;0.05) were significantly increased in liver slices collected from MCU KO mice compared to WT mice, and were associated with reductions in hepatic triacylglycerol content in vivo (p&amp;lt;0.01). Hepatocellular redox state was also altered with MCU KO, as indicated by a decreased cytosolic redox ratio ([lactate]:[pyruvate]; p&amp;lt;0.05) and increased mitochondrial redox ratio ([β-hydroxybutyrate]:[acetoacetate]; p&amp;lt;0.01). Conclusion: Here we show that increases in [Ca2+]cyt potently increase hepatic mitochondrial fat oxidation and alters hepatic redox balance to reduce hepatic lipid content independently of [Ca2+]mt in vivo and targeting [Ca2+]cyt may represent a novel therapeutic target to treat NAFLD. Disclosure T.E.Lamoia: None. B.T.Hubbard: None. M.Guerra: None. R.Goodman: None. M.H.Nathanson: None. G.I.Shulman: Consultant; Novo Nordisk, DiCerna Pharmaceuticals, Inc., Bayer Consumer Care AG, Kriya Therapeutics, Arrowhead Pharmaceuticals, Inc., ESPERION Therapeutics, Inc., Other Relationship; AstraZeneca, Merck &amp; Co., Inc., Janssen Research &amp; Development, LLC, iMetabolic Biopharma Corporation, Maze Therapeutics, 89bio, Inc., Equator Therapeutics, Inc., Generian Pharmaceuticals, Fortress Biotech, Inc., OrsoBio, Aro Biotherapeutics Company, Stock/Shareholder; Levels Health, Inc. Funding National Institutes of Health (F31DK126362, T32GM007324)

1571-P: Plasma FGF5 Concentration Correlates Better with Hepatic Steatosis, Blood Glucose, and Specific Plasma Free Fatty Acids than with Other Tissue Fat Accumulation

A few studies reported an association between fibroblast growth factor-5 (FGF5) and NAFLD. We aimed to elucidate the physiologic implication of FGF5 in NAFLD and obesity. A total of 180 subjects [(24 healthy controls and 156 patients with biopsy-proven NAFLD and/or hepatic fat content (HFC) 5% or more on MRI-PDFF] were included to assess association between HFC and various metabolic factors including body fat and plasma levels of glucose, insulin, FGF5, FGF21, oxyntomodulin, incretins, and some free fatty acids (FFAs). Additionally, we analysed a subgroup of obese patients who would undergo laparoscopic sleeve gastrectomy (LSG) (n=50) to see correlation between changes in metabolic variables related to body weight (BW) and HFC before and 6 months after surgery. Plasma FGF5 correlated better with blood glucose and HFC than with total body fat, HOMR-IR, and visceral fat. In relation to plasma FFAs, both FGF5 and HFC together showed a positive correlation with plasma FFAs such as palmitic acid, linolenic acid, and arachidonic acid, but had an inverse correlation with nervonic acid, docosanoic acid, tetracosanoic acid, and hexacosanoic acid. FGF5 level showed an independent association with blood glucose and MRI-PDFF after adjusting for multiple factors. After LSG, BW decreased by an average of 24.6%, while HFC decreased from 19.5% ± 9.1% to 5.7 ± 2.9%, p &amp;lt;0.01). Plasma FGF5 also decreased after LSG (1625.0 ± 656.8 pg/ml vs. 966.8 ± 339.9 pg/ml, p &amp;lt;0.01). The amount of plasma FGF5 change showed a significant correlation with changes in HFC and blood glucose. In conclusion, our results showed that plasma FGF5 level correlated well with HFC, blood glucose, and specific plasma FFAs and also decreased with the improvement of hepatic steatosis after bariatric surgery. Further researches on the causal relationship between FGF5, glucose and lipid metabolism, and NAFLD are needed. Disclosure D.Lee: None. I.Park: None. Funding National Research Foundation of Korea (2021R1A5A2030333, 2019R1I1A2A02062305)

370-OR: Optimized Weight Loss with Bimagrumab—Reduced Fat Mass with Increased Muscle Mass by Appetite-Independent Mechanisms

Background: Obesity is managed with lifestyle changes or incretin Tx to produce wt loss that is challenging to maintain long-term, perhaps because lean mass loss accounts for 20-40% of total wt loss. Semaglutide Tx results in rapid wt loss with 35% lower caloric intake, no further wt loss after 1 yr and rapid wt regain post-Tx. Bimagrumab (bima), a mAb to ActRIIA/ActRIIB, blocks activin pathways, leading to increased lean mass/decreased fat mass in numerous clinical studies (N=1561). Effects of bima IV or SC on fat and muscle were evaluated by DXA and MRI. Results: In a Ph2 study of bima 10 mg/kg IV q4wks vs placebo (respectively) in persons with overweight/obesity and T2DM (N=75), LS mean change at Wk 48 in body wt was −5.9 vs −0.8 kg (p&amp;lt;0.001; −7.5% vs −1.1%), with minimal change in daily caloric intake. DXA: LS mean change at Wk48 in total body (TB) fat mass was −7.5 vs −0.2 kg (p&amp;lt;0.001; −22.2% vs −0.3%), TB lean mass +1.7 vs −0.4 kg (p&amp;lt;0.001; +3.6% vs -0.9%), and appendicular lean mass +0.5 vs −0.3 kg (p=0.009; +2.5% vs −1.1%); fat mass loss was maintained 12 wks post-last dose. MRI: LS mean change at Wk24 in visceral adipose tissue was −1.5 vs +0.2 L (p&amp;lt;0.001; −36.1% vs +3.6%), absolute hepatic fat fraction (FF, %) −4.6 vs +0.2 (p=0.006; -23.6% vs +14.3%), and paravertebral muscle (PVM) mass +363.3 vs −331.0 mm2 (p=0.003; +6.4% vs -4.6%); a trend for decreased PVM fat fraction and intermuscular adipose tissue was observed (p=NS). Bima improved HbA1c (%) by -0.76 vs +0.04. AEs included diarrhea, muscle spasms. In a Ph1 study comparing bima 525 mg SC (N=12) and 700 mg IV (N=8), q4wks X3, fat mass loss and lean mass (total body and appendicular) increase were similar, with fat mass loss maintained 12 wks post-last dose; SC arm had fewer AEs than IV arm. Conclusions: Bimagrumab Tx results in decreased fat deposition in tissues and increased muscle mass despite minimal change in caloric intake, which could lead to durable, high quality wt loss. A study of bima in obesity without T2DM is ongoing (NCT05616013). Disclosure K.M.Attie: Employee; Versanis Bio. S.B.Heymsfield: None. L.Mindeholm: Consultant; Versanis Bio, Novartis AG, Stock/Shareholder; Novartis AG, Alcon Research, LLC, Novo Nordisk A/S. S.Spruill: None. M.Pruzanski: None. L.Klickstein: None.

801-P: Short-Acting Exenatide Added Three Times Daily to Insulin Therapy Improves Fatty Liver Index in Type 1 Diabetes

Fatty liver index (FLI), a marker of hepatic fat content, has been reported as an independent risk factor for all-cause mortality and major cardiovascular events in type 1 diabetes. How short-acting glucagon-like peptide 1 receptor agonists (GLP-1RA) influence FLI in type 1 diabetes remains unknown. We analyzed FLI post-hoc in a phase 2, double-blind, randomized (1:1), clinical trial (NCT03017352) that assessed the efficacy of short-acting GLP-1RA exenatide 10 μg added three times daily to insulin therapy as compared with placebo over 26 weeks of treatment in 108 individuals with type 1 diabetes (76/32 men/women, age 50 (14) years (mean (standard deviation)), diabetes duration 21 (12) years, BMI 28 (4) kg/m2, glycated hemoglobin A1c 8.2 (0.7) % / 66 (7) mmol/mol). Exenatide significantly reduced FLI by 23 % (95 % confidence interval -44; -10, P = 0.001) (Table 1) as compared with placebo after 26 weeks’ treatment. In conclusion, short-acting exenatide reduced FLI-assessed hepatic fat content in patients with long-standing type 1 diabetes and inadequate glycemic control, alluding to potential beneficial effects of this GLP-1RA on both morbidity and mortality in type 1 diabetes. Disclosure N.Johansen: Other Relationship; AstraZeneca, Novo Nordisk, Research Support; JDRF. T.F.Dejgaard: Advisory Panel; Novo Nordisk, Research Support; Novo Nordisk, Speaker's Bureau; Boehringer-Ingelheim, Novo Nordisk, AstraZeneca. A.Lund: Speaker's Bureau; Novo Nordisk. C.K.Nielsen: None. T.Vilsbøll: Consultant; AstraZeneca, Boehringer Ingelheim Inc., Gilead Sciences, Inc., Eli Lilly and Company, Mundipharma, Merck &amp; Co., Inc., Novo Nordisk A/S, Sanofi, Sun Pharmaceutical Industries Ltd., Bristol-Myers Squibb Company. F.K.Knop: Advisory Panel; AstraZeneca, Boehringer Ingelheim International GmbH, Eli Lilly and Company, Novo Nordisk, Sanofi, Consultant; AstraZeneca, Boehringer Ingelheim International GmbH, Eli Lilly and Company, Novo Nordisk, Sanofi, Research Support; Novo Nordisk, Zealand Pharma A/S, Speaker's Bureau; AstraZeneca, Boehringer Ingelheim International GmbH, Eli Lilly and Company, Novo Nordisk, Sanofi, Lundbeck. Funding AstraZeneca

244-OR: Signs of Glucagon Resistance after a Two-Week Hypercaloric Diet Intervention

Hyperglucagonemia is observed in individuals with obesity and contributes to the hyperglycemia of persons with diabetes. Hyperglucagonemia may arise due to reduced hepatic amino acid turnover and ensuing elevations in glucagonotropic amino acids. We recently observed reduced glucagon sensitivity and hyperglucagonemia in individuals with obesity and hypothesized it to be a consequence of steatosis. Here, we evaluated whether reduced glucagon sensitivity could be induced by a short-term hypercaloric diet intervention designed to increase hepatic fat content in healthy individuals. We recruited 20 healthy, lean individuals (BMI: 23 ± 0.3 (mean ± SD) kg/m2) to follow a hypercaloric diet (~5,000 kcal/day) and a sedentary lifestyle for 2 weeks. Amino acid turnover in response to infusion of glucagon was assessed during a pancreatic clamp with somatostatin and basal insulin. Participants were examined before and after the lifestyle-intervention and again after 8 weeks to assess the reversibility of any metabolic changes. Hepatic steatosis was assessed by magnetic resonance spectroscopy. The intervention led to increases in body weight (3.5 [2.8;4.2] (mean [95% confidence interval]) kg, P &amp;lt; 0.0001) and hepatic fat content (382 [206;705]%, P &amp;lt; 0.001). Insulin resistance was evident with unchanged fasting blood glucose levels and a 56% increase in fasting insulin (P &amp;lt; 0.001). Glucagon infusion led to a decrease in the concentration of total amino acids, but the percentage change in total amino acids was reduced (−2.5 ± 0.5 vs. −0.2 ± 0.7%, P = 0.015) and the average slope of the total amino acid curve was less steep (−2.0 ± 1.2 vs. −1.2 ± 0.3 μM/min, P = 0.016) after the intervention compared to baseline. All metabolic changes were normalized at follow-up. Our results indicate that short-term unhealthy behavior increasing hepatic fat content causes a reversible resistance to the effect of glucagon on amino acid turnover in healthy individuals, which may explain hyperglucagonemia associated with obesity and diabetes. Disclosure M.P.Suppli: None. J.I.Bagger: Speaker's Bureau; Novo Nordisk A/S. J.J.Holst: Advisory Panel; Novo Nordisk A/S, Eli Lilly and Company, Board Member; Bainan Biotech, Antag Therapeutics, Consultant; Alphasights, Eli Lilly and Company, ShouTi Pharma Inc., Zealand Pharma A/S, Other Relationship; Novo Nordisk, Novo Nordisk Pharma, Mayo Clinic, Boehringer-Ingelheim, Scohia Pharma Inc., Research Support; ARLA, European Union, Novo Nordisk Foundation. F.K.Knop: Advisory Panel; AstraZeneca, Boehringer Ingelheim International GmbH, Eli Lilly and Company, Novo Nordisk, Sanofi, Consultant; AstraZeneca, Boehringer Ingelheim International GmbH, Eli Lilly and Company, Novo Nordisk, Sanofi, Research Support; Novo Nordisk, Zealand Pharma A/S, Speaker's Bureau; AstraZeneca, Boehringer Ingelheim International GmbH, Eli Lilly and Company, Novo Nordisk, Sanofi, Lundbeck.

1252-P: Differences in Cardiometabolic and Immune Biomarkers between Novel Subphenotypes of Individuals at Elevated Risk for Type 2 Diabetes

Recent efforts to address heterogeneity among individuals at elevated risk for type 2 diabetes (T2D) have focused on metrics derived from oral glucose tolerance tests, body fat distribution, and hepatic fat content to subphenotype and stratify patients. A novel classification scheme identified six subphenotypes with significantly distinct risks of developing T2D, complications of insulin resistance, and mortality. The aim of our study was to evaluate differences in cardiometabolic and immune biomarkers among these clusters. Fasting serum levels of 5 Olink targeted protein panels (Cardiovascular III, Cardiometabolic, Immune response, Inflammation, Metabolism) comprising 460 oligo-species were measured in 1,597 individuals without diabetes from the Bialystok PLUS study and the Polish Registry of Diabetes. Pairwise differences between subgroups for biomarkers were assessed with generalized linear mixed models, with and without adjustment for the clustering variables. In the unadjusted model, 164 biomarkers showed significantly different levels among clusters (q-value &amp;lt; 0.05), whereas the adjusted model identified 166 significantly different biomarkers. Generally, cluster 2 (lowest T2D and mortality risk) had a more favorable profile of cardiometabolic and immune biomarkers, with clusters 5 and 6 (high-risk) showing more unfavorable profiles. Among significant biomarkers, the greatest differences were observed for tissue-type plasminogen activator and fatty acid binding protein 4, with lowest levels in cluster 2 and highest levels in clusters 5 and 6. Insulin-Like Growth Factor Binding Protein 1 and Paraoxonase 3 showed the greatest differences in the opposite direction. Our study shows that clusters of individuals at elevated risk for T2D stratify across orthogonal axes of cardiometabolic and immune biomarkers, underlining a prominent independent role of these pathways in the pathophysiology of novel subtypes. Disclosure L.Szczerbinski: None. P.Sowa: None. I.Kowalska: None. K.A.Kaminski: Other Relationship; AstraZeneca, Boehringer Ingelheim Inc. J.C.Florez: Consultant; AstraZeneca, Novo Nordisk, Other Relationship; AstraZeneca, Merck &amp; Co., Inc. A.J.Kretowski: None. A.Janucik: None. W.Kwedlo: None. P.Konopka: None. A.Citko: None. A.Paszko: None. M.Taylor: None. M.Czajkowski: None. M.Kondraciuk: None.

Taurine Protects against the Fatty Liver Hemorrhagic Syndrome in Laying Hens through the Regulation of Mitochondrial Homeostasis.

Metabolic-associated fatty liver disease (MAFLD) is a chronic liver disease caused by fat deposition in the liver of humans and mammals, while fatty liver hemorrhagic syndrome (FLHS) is a fatty liver disease in laying hens which can increase the mortality and cause severe economic losses to the laying industry. Increasing evidence has shown a close relationship between the occurrence of fatty liver disease and the disruption of mitochondrial homeostasis. Studies have proven that taurine can regulate hepatic fat metabolism, reduce hepatic fatty deposition, inhibit oxidative stress, and alleviate mitochondrial dysfunction. However, the mechanisms by which taurine regulates mitochondrial homeostasis in hepatocytes need to be further studied. In this study, we determined the effects and mechanisms of taurine on high-energy low-protein diet-induced FLHS in laying hens and in cultured hepatocytes in free fatty acid (FFA)-induced steatosis. The liver function, lipid metabolism, antioxidant capacity, mitochondrial function, mitochondrial dynamics, autophagy, and biosynthesis were detected. The results showed impaired liver structure and function, mitochondrial damage and dysfunction, lipid accumulation, and imbalance between mitochondrial fusion and fission, mitochondrial autophagy, and biosynthesis in both FLHS hens and steatosis hepatocytes. Taurine administration can significantly inhibit the occurrence of FLHS, protect mitochondria in hepatocytes from disease induced by lipid accumulation and FFA, up-regulate the expression levels of Mfn1, Mfn2, Opa1, LC3I, LC3II, PINK1, PGC-1α, Nrf1, Nrf2, and Tfam, and down-regulate the expression levels of Fis1, Drp1, and p62. In conclusion, taurine can protect laying hens from FLHS through the regulation of mitochondrial homeostasis, including the regulation of mitochondrial dynamics, autophagy, and biosynthesis.

1599-P: Regulation of Mitochondrial Dynamics Ameliorates Hepatic Steatosis through TFEB Activation

Nonalcoholic fatty liver disease (NAFLD) is a common cause of liver disease that is closely related to metabolic syndromes, obesity, and type 2diabetes. There is strong evidence that mitochondrial dysfunction plays a significant role in the development and progression of NAFLD. Recently, growing evidence suggests that the TFEB plays a pivotal role in mitochondrial dynamics. In this study, we investigated that mitochondrial dynamics play the potential role and mechanisms of TFEB in the amelioration of NAFLD. FFA increased expression of mitochondrial fission proteins p-Drp1, Fis1 and decreased expression of fusion proteins OPA1, Mfn2 and inhibition of TFEB activation. Mitotracker stain significantly increased in the proportion of cells with fragmented mitochondria in FFA-treated cells. HFD-fed mice displayed histology consistent with pathological hepatic steatosis. EM analysis indicated that HFD-fed mitochondria tended to be shorter and more round than ND-fed mitochondria. Western blot analyses reveal increased expression of the fission proteins, p-Drp1, Fis1, and significantly decreased in the fusion protein, Mfn2 and OPA1 in HFD-fed group. In vivo Tfeb knockout dramatically increased hepatic fat accumulation, liver ballooning in HFD-fed mice. EM analysis showed that HFD-fed Tfeb knockout mice had more rounded and swollen mitochondria, indicating that TFEB was dependent on the regulation of mitochondrial fragmentation. In vivo Tfeb knockout reveal significantly increased expression of the fission proteins in HFD-fed group. To demonstrated the direct regulatory mechanism of mitochondrial dynamics and TFEB activation, we used siRNA DRP1 and Mdivi-1. Inhibition of Drp1 promotes mitochondrial fusion, increased expression of fusion protein and reduced fat accumulation and activated TFEB. These results suggest a role in improving hepatic steatosis through regulation of TFEB-mediated mitochondrial dynamics in NAFLD. Disclosure J.Yoo: Other Relationship; American Diabetes Association. Y.Hwang: None. Funding National Research Foundation of Korea (2022R1I1A1A01069899)

166-LB: Clinical Improvements and Extracellular Vesicle Size following Lifestyle Intervention in Latino Youth with Nonalcoholic Fatty Liver Disease

Latino youths with obesity are disproportionately impacted by nonalcoholic fatty liver disease (NAFLD). Lifestyle intervention can improve established biomarkers such as serum transaminases and hepatic fat fraction (HFF) in NAFLD patients. Circulating levels of extracellular vesicles (EVs) have been associated with NAFLD and may track with changes in liver fat. Here we assessed changes in EV concentration and size following a comprehensive lifestyle intervention in 18 Latino adolescents (12 males/6 females; average age 13.3 ±1.2y) with obesity and NAFLD, as defined by MRI-measured hepatic fat fraction (HFF) ≥5%. Participants completed a 6-month lifestyle intervention that targeted improvements in eating behaviors (e.g., decreased sugar and saturated fat intake, portion control, and increased fiber intake) and increases in physical activity. EVs were isolated from fasting plasma samples taken at baseline and following intervention using size exclusion chromatography and characterized by nanoparticle tracking analysis. Significant improvements (pre- vs post-intervention) were observed in percent body fat (44.7 ±4.8% vs 42.2 ±5.3%, p&amp;lt;0.0001), alanine transaminase (ALT; 24.8 ±7.4 U/L vs 23.2 ±5.6 U/L p=0.02), and HFF (12.5 ±5.5% vs 9.6 ±4.9%, p=0.008). We observed a significant reduction in mean EV size (128.4 ±16.5 vs 120.2 ±16.4, p=0.03) and a non-significant reduction in EV concentration (1.14E+09 ±4.1E+08 vs 1.09E+09 ±1.8E+08, p=0.61) following the intervention. Change in mean EV size was significantly and positively associated with changes in SAT (cm3), HDL (mg/dl), fat (kg) and BMI (all p &amp;lt; 0.05). These results suggest that improvements in NAFLD-related phenotypes following lifestyle intervention correspond with reductions in EV size in Latino adolescents with obesity and NAFLD. Disclosure I. Piras: None. X. Wu: None. P. Pirrotte: None. M. Olson: Advisory Panel; Rhythm Pharmaceuticals, Inc. S. Khan: None. S. Xanthakos: None. G. Q. Shaibi: None. J. Distefano: None. Funding National Institutes of Health (R01DK127015)

PLAAT1 deficiency alleviates high-fat diet-induced hepatic lipid accumulation in mice.

The phospholipase A and acyltransferase (PLAAT) family is composed of three isoforms in mice (PLAAT1, 3, and 5), all of which function as phospholipid-metabolizing enzymes exhibiting phospholipase A1 /A2 and acyltransferase activities. Plaat3-deficient (Plaat3-/- ) mice were previously reported to show lean phenotype and remarkable hepatic fat accumulation under high-fat diet (HFD) feeding, while Plaat1-/- mice have not been analyzed. In the present study, we generated Plaat1-/- mice and investigated the effects of PLAAT1 deficiency on HFD-induced obesity, hepatic lipid accumulation, and insulin resistance. After HFD treatment, PLAAT1 deficiency caused a lower body weight gain compared to wild-type mice. Plaat1-/- mice also showed reduced liver weight with negligible hepatic lipid accumulation. In accordance with these findings, PLAAT1 deficiency improved HFD-induced hepatic dysfunction and lipid metabolism disorders. Lipidomics analysis in the liver revealed that in Plaat1-/- mice, the levels of various glycerophospholipids tended to increase, while all classes of lysophospholipids examined tended to decrease, suggesting that PLAAT1 functions as phospholipase A1 /A2 in the liver. Interestingly, the HFD treatment of wild-type mice significantly increased the mRNA level of PLAAT1 in the liver. Furthermore, the deficiency did not appear to elevate the risk of insulin resistance in contrast to PLAAT3 deficiency. These results suggested that the suppression of PLAAT1 improves HFD-induced overweight and concomitant hepatic lipid accumulation.

Which obesity index is the most useful marker for predicting hepatic steatosis in children and adolescents with obesity? A cross-sectional study using quantitative magnetic resonance imaging

IntroductionWe aimed to evaluate the relationships between hepatic steatosis and various indices of obesity, and to identify the most useful index for the prediction of hepatic steatosis in children and adolescents with obesity. MethodsA total of 226 children and adolescents with a mean body mass index (BMI) z-score of 2.65 and a mean age of 11.4 years were subjected to anthropometric and body composition measurements, laboratory testing, abdominal fat mass assessment, and hepatic fat accumulation by magnetic resonance imaging-derived proton density fat fraction (MRI-PDFF). The participants were divided into quartiles according to the severity of their hepatic steatosis, and the presence of hepatic steatosis was defined using an MRI-PDFF ≥ 5%. ResultsThe multivariate ordinal regression analysis showed that the severity of hepatic steatosis was positively associated with BMI, waist circumference, waist-to-hip ratio, waist-to-height ratio, fat mass, fat-free mass, visceral adiposity, and abdominal subcutaneous adiposity. Higher activities of liver enzymes and higher concentrations of triglyceride, C-reactive protein, fasting insulin, and leptin were associated with more severe hepatic steatosis, whereas high-density lipoprotein-cholesterol and adiponectin were negatively associated with hepatic steatosis. The indices of obesity with areas under the receiver operating characteristic curves (AUCs) > 0.8 for the prediction of hepatic steatosis were liver enzymes, visceral adipose tissue area, waist-to-hip ratio, and waist-to-height ratio. ConclusionThe severity of hepatic steatosis significantly correlated with various indices of obesity and cardiometabolic markers in children and adolescents with obesity. The indices of abdominal obesity would be the most useful for the prediction of hepatic steatosis.

Hepatic Fat and Macrophages Are Increased in Livers of Diabetic Patients without Non-Alcoholic Fatty Liver Disease

Introduction: Diabetes mellitus (DM), especially type 2, is strongly associated with non-alcoholic fatty liver disease (NAFLD). Recent studies indicate that particularly in DM patients, “simple” liver steatosis can progress into more severe disease. However, little is known about putative hepatic histopathological changes in DM patients without NAFLD. In this study, we therefore analysed fat content and inflammatory cell infiltration in the livers of deceased DM and non-DM patients without NAFLD, and analysed age/sex effects hereon. Methods: Hepatic fat and inflammatory cells were studied through (immuno)histochemical analysis in liver tissue from 24 DM patients and 66 non-diabetic controls, without histopathological characteristics of NAFLD. Results: We observed a 2-fold increase in fat percentage/mm² and a near 5-fold increase in the number of fat-containing cells/mm² in DM patients compared to non-diabetic controls. Fat content was significantly higher in patients with type 2 DM, but not type 1 DM, compared to non-diabetic controls, while the number of CD68+ cells/mm² was significantly elevated in both DM groups. Conclusion: Hepatic fat and number of macrophages are increased in patients with DM without NAFLD, which may reflect a higher risk on development of steatosis and steatohepatitis.