Liver Fat Content Research Articles

98-OR: Oral Insulin Alleviates Liver Fibrosis and Reduces Fat in Patients with NASH and Type 2 Diabetes—Results of Phase II Clinical Trial

NAFLD is the most common liver disorder in the USA. NASH is an advanced form of NAFLD and a leading cause of end-stage liver disease and indication for liver transplantation. Insulin resistance and inflammation underlie the pathogenesis of the disease. This double-blind, randomized, placebo-controlled, multicenter study aimed to determine the effect of oral insulin in patients with NASH and T2DM. Patients were treated twice daily for 12 weeks with an 8 mg insulin (n=21) or placebo (n=11) capsule. Safety was monitored throughout the study. At screening and after 12 weeks of treatment, liver fat content was measured by MRI-PDFF and liver fibrosis and steatosis levels by FibroScan®. No severe drug-related adverse events were reported during the study. After 12 weeks of treatment, mean percent reductions in whole-liver (-11.2±22.8% vs. -6.5±15.3%, respectively) and liver segment 3 (-11.7±21.1% vs. +0.1±17.3%, respectively) fat content were higher in the insulin as compared to the placebo arm. Patients receiving insulin showed a median -1.1 kPa and -21.0 dB/m reduction from baseline fibrosis and steatosis levels, respectively. In comparison, placebo-treated patients showed median increases of 0.3 kPa and 13.0 dB/m, respectively. At Week 12, HbA1c levels decreased by a mean -0.27±1.3% in the oral insulin cohort and increased by a mean 0.23±0.73% in the placebo cohort. Mean percent changes from baseline ALT and AST levels were -0.1±30.4% and -0.8±28.6%, respectively, in the oral insulin arm and 3.0±26.7% and 13.4±46.6%, respectively, in the placebo arm. Over the treatment period, mean total cholesterol, HDL, LDL, and triglycerides levels were consistently lower than baseline in the active arm and higher than baseline in the placebo arm. The results of this feasibility study support the safety and potential therapeutic effect of orally delivered insulin on liver fibrosis, fat accumulation, and inflammatory processes. Disclosure Y.Ishay: None. J.M.Neutel: None. Y.Kolben: None. R.Gelman: None. O.Sneh arbib: None. O.Lopez: None. R.Sotolongo: None. Y.Ilan: None. M.Kidron: Employee; Oramed Pharmaceuticals.

191-OR: Deletion of the Type 2 Diabetes Candidate Gene SLC16A11 Reduces Peripheral Insulin Sensitivity in Mice

Background and Aims: Recent GWAS in large human populations identified the monocarboxylate transporter SLC16A11 as a candidate gene for T2D. While previous studies postulated that SLC16A11 plays a role in hepatic lipid and glucose metabolism, the existing data are controversial and exact mechanisms of how SLC16A11 affects T2D are missing. Here, we aimed at elucidating the role of SLC16A11 in the development of metabolic disease. Methods: HEK293 cell lines expressing murine and human SLC16A11 were established. Moreover, using CRISPR/Cas9, a SLC16A11 knockout mouse model was generated. Finally, SLC16A11 mRNA expression was measured in various insulin responsive tissues of ob/ob and db/db mice. Results: In HEK293 cells, substrates of the transporter were identified. mRNA expression of SLC16A11 was reduced in skeletal muscle of ob/ob and db/db mice (-69.8% and -22.8%; p<0.05) compared to littermate controls. SLC16A11 KO and WT littermate mice fed NCD or HFD did not differ in body weight and composition, energy expenditure or food intake. While basal glucose and insulin in SLC16A11 KO and WT littermates on a HFD were not different, glucose infusion rate during hyperinsulinemic euglycemic clamp was lower in SLC16A11 KO mice (SLC16A11 WT= 36.4±3 mg/kg/min; SLC16A11 KO= 24.7±2 mg/kg/min; P<0.01), indicating reduced whole-body insulin sensitivity. No changes were observed in hepatic glucose production, liver fat content or hepatic mitochondrial function using PINTA method. Instead, clamped 2-deoxy-glucose uptake was significantly reduced in skeletal muscle (-41%; p<0.05) of SLC16A11 KO, together with increased DAGs levels and PKCθ activation, indicating skeletal muscle insulin resistance. Conclusions: Our data show for the first time that in contrast to previous assumptions, SLC16A11 impairs skeletal muscle insulin sensitivity without affecting hepatic metabolism. Taking also human GWAS into account, we propose that SLC16A11 may be an interesting target for the treatment of T2D. Disclosure N.N.El-agroudy: None. T.Schumann: None. A.Kurzbach: None. G.Sancar: None. L.Sandforth: None. C.Herrmann: 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 & Co., Inc., Janssen Research & 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. A.L.Birkenfeld: None.

220-LB: Characterizing the Role of 99 Cardiometabolic Candidate Genes Using CRISPR/Cas9, In Vivo Imaging, and Deep Learning in Zebrafish Larvae

Introduction: Genome-wide association studies identified 100s of loci that are associated with the risk of common cardio-metabolic diseases (T2D and CVD). Here we functionally characterise 99 candidate genes for a role in relevant traits using zebrafish larvae. Methods: Fifty-six T2D and 46 CVD candidate genes (three overlap) were functionally characterised by targeting their zebrafish orthologs using CRISPR/Cas9. On day 9-11 post-fertilisation, we acquired optical sections of the pancreatic islet, liver and vasculature using semi-automated fluorescence microscopy, in 16,141 CRISPR/Cas9-edited zebrafish larvae, followed by image analysis using deep learning. Results: Perturbing all nine T2D genes with at least moderate prior evidence affects ≥1 T2D trait (i.e., beta cell mass, beta cell insulin expression, liver fat, and/or glucose content), while no consequence is observed for 22 genes without prior evidence of a role in T2D. For 12 genes with at most modest prior evidence, perturbation also affects at least 1 T2D trait. Perturbation of 23 genes prioritised for a role in CVD affects at least one vascular trait, while no consequence is observed for 15 non-prioritised genes. Effects of mutations in mice and/or humans are published for 14 of 23 genes, 78% of which show directionally consistent effects across zebrafish larvae and mammals. Of the 15 genes influencing liver fat upon CRISPR/Cas9 editing, mutations in 12 previously showed an effect on liver fat in mice and/or humans, with 75% congruence in direction of effect across species. Conclusion: Systematically characterising candidate genes for a role in image-based T2D, liver fat and CVD traits in CRISPR/Cas9-edited zebrafish larvae shows highly congruent results with mice and humans, and prioritises promising genes for further in-depth characterisation. Disclosure E. Mujica: None. H. Sun: None. M. Nyberg: Employee; Novo Nordisk A/S. D. Djordjevic: Employee; Novo Nordisk. S. Vienberg: Employee; Novo Nordisk A/S. A. L. Gloyn: Other Relationship; Genentech, Inc., Roche Pharmaceuticals. A. Larsson: None. A. Allalou: None. M. Den hoed: None. H. Zhang: None. A. Emmanouilidou: None. E. Mazzaferro: None. N. Cook: None. C. Metzendorf: None. G. Alavioon: None. M. Bandaru: None. A. Rottner: Employee; AstraZeneca. Funding Swedish Research Council (201901417); Swedish Heart and Lung Foundation (20200602, 20200781)

1325-P: Continuous Representation of Type 2 Diabetes Heterogeneity on a Tree-Like Graph Structure Stratifies Risk of Complications

Heterogeneity in type 2 diabetes (T2D) can be represented by a tree-like structure using reverse graph-embedded dimensionality reduction. We aimed to validate this approach and study if intensified phenotyping stratifies complications. Participants (n=927) of the German Diabetes Study (GDS) with recent-onset T2D were mapped onto a two-dimensional tree based on HbA1c, BMI, total and HDL cholesterol, triglycerides, alanine aminotransferase, creatinine, systolic and diastolic blood pressure, residualized for age and sex. A subset (n=447) had follow-up examinations after 5 years. Phenotypic variability rendered on a tree structure showed gradients for insulin sensitivity (hyperinsulinemic-euglycemic clamp, both dimensions: p<0.001) and insulin secretion (intravenous glucose tolerance test, both dimensions: p<0.001) (Fig 1). Individuals in the tree branch with the lowest insulin sensitivity had the highest liver fat content (1H-MR spectroscopy, pdim1<0.001, pdim2=0.04), elevated cardiovascular hazard (nevents=147, HRmax 4.0, pdim1=0.2, pdim2<0.001) and diabetic retinopathy (nevents=36, fundus photography, HRmax 2.5, pdim1=0.04, pdim2=0.2). This method provides an alternative approach to discrete clusters in identifying persons at high risk of certain diabetes-related diseases, potentially providing a framework for precision diabetology. Disclosure M.Schön: None. G.J.Bönhof: None. A.L.Birkenfeld: None. J.Seissler: None. A.F.Pfeiffer: Advisory Panel; Abbott Diabetes, Speaker's Bureau; Novo Nordisk, Sanofi-Aventis Deutschland GmbH. M.Blüher: Advisory Panel; Boehringer Ingelheim Inc., Lilly, Novo Nordisk, Consultant; Novo Nordisk Foundation, Speaker's Bureau; Amgen Inc., AstraZeneca, Bayer Inc., Daiichi Sankyo, Novartis, Sanofi-Aventis Deutschland GmbH. S.R.Bornstein: None. J.Szendroedi: None. S.Meyhoefer: None. V.Burkart: None. V.Schrauwen-hinderling: None. K.Prystupa: Other Relationship; Berlin-Chemie AG. O.Kuss: Consultant; Berlin-Chemie AG. E.Pearson: Speaker's Bureau; Novo Nordisk, Lilly, Illumina. M.Roden: Advisory Panel; Eli Lilly and Company, Consultant; TARGET PharmaSolutions, Inc., Research Support; Boehringer-Ingelheim, Novo Nordisk, Novartis, Sanofi. R.Wagner: Advisory Panel; Daiichi Sankyo, Speaker's Bureau; Novo Nordisk, Sanofi. The german diabetes study (gds) group: n/a. T.Mori: None. O.P.Zaharia: None. K.Bódis: None. Y.Kupriyanova: None. A.Nair: None. A.Strom: None. R.Guthoff: Other Relationship; Alimera, Bayer Inc., Novartis, Allergan. Funding German Diabetes Center; German Federal Ministry of Health (Berlin, Germany); Ministry of Culture and Science of the state North Rhine-Westphalia (Düsseldorf, Germany); German Federal Ministry of Education and Research; German Center for Diabetes Research

728-P: Biomarkers of Low-Grade Inflammation in Novel Subtypes of Patients at Risk for Diabetes

In the Tübingen Family Study recently 6 subtypes of prediabetic metabolism were identified. Cluster 3, 5 and 6 have a moderate to high risk for T2D or its associated complications. Cluster 3 is characterized by low insulin secretion, cluster 5 by insulin resistance and high liver fat content. Individuals in cluster 6 have an elevated risk of nephropathy despite a delayed diabetes development. Low-grade inflammation has been implicated in the development of T2D and cardiorenal complications. We hypothesized that the subtypes differ in levels of inflammatory biomarkers. Thus, we measured 31 inflammatory biomarkers associated with metabolic disease. We used Bio-Plex® multiplex immunoassays in 145 individuals who were recruited in Tübingen from the Prediabetes Lifestyle Intervention Study (PLIS) - a multicenter study conducted in individuals with ADA-defined prediabetes. Cluster comparison was done using ANOVA models, raw unadjusted values are reported. We identified eight biomarkers which differed among diabetes-risk subtypes, namely MIP1a, MIP1b, IL18, Osteopontin (OPN), IFNy, TNFb, and soluble TNFa-Receptor (sTNF-R)- 1 and -2. High risk cluster 3, 5 and 6 had significantly elevated levels of several inflammatory biomarkers compared to low risk cluster 2 with the lowest levels of these biomarkers. Within the high-risk groups, cluster 5 had increased biomarker levels compared to cluster 3 (e.g., IL-18: cluster 5: 86.2 pg/ml ± 3.9 vs cluster 3: 66.8 pg/ml ± 4.8, p=0.018; OPN: 16.8 ng/ml ± 1.0 vs. 12.8 ng/ml ± 0.8, p = 0.022; MIP1a 2.2 pg/ml ± 0.1 vs. 1.6 pg/ml ± 0.1, p < 0.001). Cluster 6 showed lower levels of IFNy than cluster 3 (2.2pg/ml ± 0.3 vs 3.5pg/ml ± 0.2, P=0.019). Our data show that the novel T2D risk subtypes differ in inflammatory biomarkers. This suggests that a low-grade inflammation may be involved in the progression from prediabetes to T2D and complications. A more precise mechanistic understanding is needed in order to establish precise preventive strategies for different risk phenotypes. Disclosure S.Katzenstein: None. J.Szendroedi: None. M.Roden: Advisory Panel; Eli Lilly and Company, Consultant; TARGET PharmaSolutions, Inc., Research Support; Boehringer-Ingelheim, Novo Nordisk, Novartis, Sanofi. L.Fritsche: None. N.Stefan: Advisory Panel; Pfizer Inc., Research Support; Sanofi, Speaker's Bureau; AstraZeneca, Boehringer Ingelheim (Canada) Ltd., Lilly Diabetes, Novo Nordisk, Sanofi-Aventis Deutschland GmbH. R.Wagner: Advisory Panel; Daiichi Sankyo, Speaker's Bureau; Novo Nordisk, Sanofi. A.Fritsche: Advisory Panel; Novo Nordisk, Lilly, Sanofi, Boehringer-Ingelheim, Speaker's Bureau; AstraZeneca, SYNLAB Holding Deutschland GmbH. A.L.Birkenfeld: None. R.Jumpertz von schwartzenberg: Other Relationship; Sanofi, Amgen Inc., Lilly, Novo Nordisk. A.Sandforth: None. V.Minelli faiao: None. J.Seissler: None. S.R.Bornstein: None. N.Perakakis: Advisory Panel; Bayer Inc., Other Relationship; Novo Nordisk, Novo Nordisk. A.Schürmann: None. S.Kabisch: Other Relationship; Berlin-Chemie AG, Sanofi, Lilly, Boehringer Ingelheim Inc., JuZo Akademie, Research Support; J. Rettenmaier / Söhne, Wilhelm-Doerenkamp Foundation, German Center of Diabetes Research. M.Blüher: Advisory Panel; Boehringer Ingelheim Inc., Lilly, Novo Nordisk, Consultant; Novo Nordisk Foundation, Speaker's Bureau; Amgen Inc., AstraZeneca, Bayer Inc., Daiichi Sankyo, Novartis, Sanofi-Aventis Deutschland GmbH.

1379-P: Adding Salt to Foods and Risk of Severe Nonalcoholic Fatty Liver

Background: High sodium intake is associated with insulin resistance, obesity, and metabolic syndrome, whereas very few studies have assessed the association of sodium intake with risk of nonalcoholic fatty liver disease (NAFLD). We recently found that the frequency of adding salt to foods could reflect a person’s long-term salt taste preference and sodium intake and was significantly related to life expectancy and cardiovascular disease risk. In this study, we aimed to evaluate the association between the frequency of adding salt to foods and liver proton density fat fraction (PDFF, a magnetic resonance imaging-based measure to assess liver fat content) and severe NAFLD risk. Methods: A total of 494,240 participants from UK biobank, who were free of NAFLD at baseline (2006-2010) were analyzed in this study. Of the above participants, 9816 participants have MRI-PDFF data, measured in 2014. Generalized linear models were used to evaluated association between the frequency of adding salt to foods with liver-PDFF; Cox proportional hazards models were used to estimate the association between the frequency of adding salt to foods and severe NAFLD risk. Results: During a median of 11 years of follow-up, 5,294 severe NAFLD cases were documented. Compared with participants who Never/Rarely added salt to foods, the adjusted Hazard Ratios (95% confidence interval) were 1.11 (1.04, 1.18), 1.27 (1.17, 1.37), and 1.45 (1.30, 1.62) across the groups of Sometimes, Usually, and Always, respectively (P-trend<0.001). The liver-PDFF values were 4.7 % (95% CI, 3.9, 5.5), 5.1% (4.3, 5.9), 5.2% (4.4, 6.0) and 5.5% (4.5-6.4) in the groups of Never/Rarely, Sometimes, Usually, and Always added salt to foods, respectively (P-trend<0.001). In sensitivity analyses, we replaced the frequency of adding salt to foods with estimated 24-hour urinary sodium extraction and found similar significant associations. Conclusions: Our findings indicate that high sodium intake is associated with higher risk of severe NAFLD. Disclosure H. Ma: None. X. Wang: None. X. Li: None. Y. Heianza: None. L. Qi: None.

838-P: Reversion of Nonalcoholic Steatohepatitis by Skeletal-Muscle Directed FGF21 Gene Therapy

Nonalcoholic steatohepatitis (NASH) is a severe form of nonalcoholic fatty liver disease, which is driven by the global obesity and type 2 diabetes epidemic. NASH is characterized by liver inflammation and fibrosis that can progress to cirrhosis, liver failure, cancer and death. Currently, NASH is an unmet medical need. Recent studies revealed the therapeutic potential of fibroblast growth factor 21 (FGF21) analogues/mimetics against NASH in pre-clinical models and in human patients, although periodic administrations are required and immunological responses may raise due to the use of engineered, non-native, proteins. Here, we demonstrated that adeno-associated viral (AAV) vector-mediated-FGF21 genetic engineering of skeletal muscle of high-fat diet-fed obese and insulin-resistant mice resulted in increased circulating levels of native FGF21. AAV-FGF21-treatment normalized liver fat content, liver inflammatory and fibrotic markers and counteracted hepatocyte damage, consistent with NASH resolution. It also precluded progression from NASH to cirrhosis and hepato-cellular carcinoma. Moreover, circulating FGF21 reversed adipose tissue hypertrophy and inflammation, increased energy expenditure, normalized glycemia and insulinemia, and increased insulin sensitivity. Scale-up of muscular FGF21 gene transfer to large animals also successfully resulted in long-term secretion of biologically active FGF21 and therapeutic effects in key target tissues, in the absence of adverse events. All these results point out the potential of the AAV-FGF21-mediated gene therapy to treat NASH and support the clinical translation of the approach. Disclosure V. Jimenez: None. V. Sacristán: None. C. Jambrina: None. M. Jaén: None. S.A. Munoz: None. E. Casana: None. M. Garcia: None. I. Grass: None. X. Leon: None. I. Elias: None. A. Ribera: None. G. Elias: None. V. Sanchez: None. A. Casellas: None. T. Ferré: None. M. Molas: None. F. Bosch: None.

Insights into the mechanism of growth and fat deposition by feeding different levels of lipid provided by transcriptome analysis of swamp eel (Monopterus albus, Zuiew 1793) liver.

Lipid is an important source of energy in fish feeds, and the appropriate fat content can improve the efficiency of protein utilization. However, excessive lipid content in the feed can lead to abnormal fat deposition in fish, which has a negative effect on the growth of fish. Therefore, the effects of feed lipid levels on swamp eel were studied. Essential functional genes were screened using transcriptomics. We divided 840 fish into seven groups (four replicates). A mixture of fish and soybean oils (1:4), 0%, 2%, 4%, 6%, 8%, 10%, and 12% was added to the basic feed were named groups one to seven (L1-L7), respectively. Isonitrogenous diets were fed swamp eel for 10 weeks. Growth performance, visceral index, nutritional components, and biochemical indexes were measured and analyzed. Livers of the 0%, 6%, and 12% groups were subjected to transcriptome sequencing analysis. The results of our study showed that: the suitable lipid level for the growth of swamp eel was 7.03%; the crude fat content of whole fish, liver, intestine, muscle, and skin increased with the increase of lipid level, with some significant difference, and excess fat was deposited in skin tissue; triglyceride, total cholesterol, and free fatty acid contents increased with the increase of feed lipid level. High-density lipoprotein levels in the L3 and L4 groups were higher than in the other groups. Blood glucose concentrations in the L5, L6, and L7 groups increased; the liver tissue structure was damaged when the lipid level was too high. two-hundred-and-twenty-eight differentially expressed genes were found. Several critical pathways regulating glucose metabolism and energy balance (e.g., glycerolipid metabolism, glycolysis synthesis, degradation of ketone bodies, and Janus Kinase/Signal Transducer and Activator of Transcription signaling pathway) were enriched in swamp eel compared with the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Suitable lipid levels (7.03%) can promote the growth of swamp eel, and excessive lipid levels can cause elevated blood lipids and lead to liver cell damage. Regulatory mechanisms may involve multiple metabolic pathways for glucose and lipid metabolism in eels. This study provides new insights to explain the mechanism of fat deposition due to high levels of lipid and provides a basis for the production of efficient and environmentally friendly feed for swamp eel.

Abdominal magnetic resonance imaging assessment and quantification of pancreatic and liver adipose tissues in obesity before and after laparoscopic sleeve gastrectomy

ObjectivesTo investigate the changes of liver and pancreas fat fraction after laparoscopic sleeve gastrectomy (LSG) by magnetic resonance imaging (MRI). MethodsOur study included morbid obesity with normal blood glucose and underwent abdominal MRI before and after LSG. In total, 21 patients were included, 10 underwent abdominal MRI before and 3 months after LSG, 11 underwent abdominal MRI before and 1–2 years after LSG. Liver fat fraction (LFF), pancreas fat fraction(PFF) were measured on proton density fat fraction (PDFF) maps at different time points. Fasting blood samples were collected for measurement of triglyceride (TG), total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL), plasma glucose and insulin levels. Homeostasis model assessment (HOMA) was used to evaluate insulin resistance (HOMA-IR). Pearson correlation was used to analyze the correlation between different parameters. ResultsLFF and PFF were significantly decreased at 3 months and 1–2 years after LSG along with weight loss, decreased body mass index (BMI) and waist circumference. Though these parameters showed a decreasing trend from 3 months to 1–2 years, but there was no statistical difference. 81.0% patients had fatty liver and 38.1% patients had fatty pancreas at baseline. However, all patients had no fatty liver at 3 months and no fatty pancreas at 1–2 years after LSG. HDL levels were increased, but LDL, TG and insulin levels were decreased 1–2 years after LSG but not at 3 months. LFF was related to fasting insulin and HOMA-IR at baseline but not after LSG. There was no association of PFF with fasting plasma glucose, fasting insulin or HOMA-IR, but ΔPFF was related to Δinsulin and ΔHOMA-IR at 1–2 years after LSG. ConclusionChanges of liver and pancreas fat content in obesity before and after LSG can be quantified by IDEAL-IQ. Treatment with LSG can reverses fatty liver and fatty pancreas in obese patients. In addition, the lipid profile and insulin sensitivity are also improved after LSG along with the decreased of LFF and PFF.

Associations of insulin-like growth factor binding protein-2 with metabolic profile and hepatic fat deposition in asymptomatic men and women.

Low circulating levels of insulin-like growth-factor binding protein-2 (IGFBP-2) have been associated with increased adiposity and metabolic alterations such as insulin resistance, dyslipidemia, and nonalcoholic fatty liver disease in individuals with obesity. However, whether IGFBP-2 affects energy metabolism in the early stages of these disorders remains unclear. Herein, we hypothesized that plasma IGFBP-2 concentrations are inversely associated with early liver fat accumulation and alterations in lipid and glucose homeostasis in apparently healthy and asymptomatic men and women. Three hundred thirty-three middle-aged Caucasian men and women apparently healthy and without cardiovascular symptoms were enrolled for a cross-sectional cardiometabolic imaging study. Individuals with BMI ≥ 40 kg/m2, cardiovascular disease, dyslipidemia, hypertension, and diabetes were excluded. Fasting glucose and lipid profiles were measured and an oral glucose tolerance test was performed. Liver fat content was assessed by magnetic resonance spectroscopy. Volume of visceral adipose tissue (VAT) was evaluated by magnetic resonance imaging. Plasma IGFBP-2 levels were quantified by ELISA. Participants with low IGFBP-2 levels were characterized by a higher body fat mass (P < 0.0001), insulin resistance (P < 0.0001), higher plasma triglyceride (TG) (P < 0.0001), and lower HDL-cholesterol levels (P < 0.0001) in a sex-independent manner. IGFBP-2 levels were inversely correlated with hepatic fat fraction in both men (r = -0.36, P < 0.0001) and women (r = -0.40, P < 0.0001). IGFBP-2 concentrations were negatively associated with hepatic fat fraction independently of age and VAT in both men (R2 = 0.23, P = 0.012) and women (R2 = 0.27, P = 0.028). In conclusion, our findings show that even in asymptomatic, apparently healthy individuals, low IGFBP-2 levels are associated with a more deteriorated cardiometabolic risk profile and with a high hepatic fat content in a VAT-independent manner. However, IGFBP-2 does not appear to influence the established sexual dimorphism observed for metabolic variables and hepatic fat fraction. Additional studies are required to better understand the relationships between IGFBP-2 and liver fat content.NEW & NOTEWORTHY Faced with a paucity of reliable clinical etiologic markers for fatty liver, this research article demonstrates, for the first time, that low blood levels of the protein IGFBP-2 are associated with a more deteriorated cardiometabolic risk profile and with a high hepatic fat content independently of visceral fat volume and sex, even in asymptomatic, apparently healthy individuals.

The impact of concomitant metabolic dysfunction-associated fatty liver disease on adverse outcomes in patients with hepatitis B cirrhosis: a propensity score matching study.

In cirrhotic patients, the clinical relevance of metabolic dysfunction-associated fatty liver disease (MAFLD) is unclear. We aimed to research the relationship between MAFLD and adverse clinical outcomes in patients with hepatitis B cirrhosis. A total of 439 patients with hepatitis B cirrhosis were enrolled. Abdominal MRI and computed tomography were used to calculate liver fat content in order to evaluate steatosis. The Kaplan-Meier method was implemented to generate survival curves. The independent risk factors for prognosis were identified by multiple Cox regression. Propensity score matching (PSM) was used to reduce the influence of confounding factors. This study explored the relevance between MAFLD and mortality, first decompensation and further decompensation. In our study, most patients were decompensated cirrhosis (n = 332, 75.6%) and the ratio of decompensated cirrhosis patients in non-MAFLD to MAFLD group was 199 : 133. Compared to the non-MAFLD group, patients with MAFLD had worse liver function which mainly reflected that there were more Child-Pugh C patients and higher model for end-stage liver disease score in the MAFLD group. A total of 207 adverse clinical events occurred in the total cohort during a median follow-up of 47 months, including 45 deaths, 28 hepatocellular carcinoma, 23 first decompensation and 111 further decompensation. Cox multivariate analysis showed that MAFLD was an independent risk factor for death [hazard ratio (HR) 1.931; 95% confidence interval (CI) 1.019-3.660; P = 0.044 HR 2.645; 95% CI, 1.145-6.115; P = 0.023] and further decompensation (HR 1.859; 95% CI, 1.261-2.741; P = 0.002 HR 1.953; 95% CI, 1.195-3.192; P = 0.008) before and after PSM. In decompensated group with MAFLD, diabetes had a more significant effect on adverse prognosis than overweight or obesity and other metabolic risk factors. In patients with hepatitis B cirrhosis, concomitant MAFLD can predict a higher risk of further decompensation and death among decompensated individuals. According to patients among MAFLD, diabetes may be a major factor in the occurrence of adverse clinical events.

Obesity management for the pre–liver transplant and post–liver transplant patient

Obesity management for the pre–liver transplant and post–liver transplant patient

OS-086 - Stratification of liver fat content in non-alcoholic steatohepatitis patients with significant liver fibrosis using the MEFIB-Index and MRI-PDFF and its association with hepatocellular carcinoma, decompensation, and mortality

OS-086 - Stratification of liver fat content in non-alcoholic steatohepatitis patients with significant liver fibrosis using the MEFIB-Index and MRI-PDFF and its association with hepatocellular carcinoma, decompensation, and mortality

Peroxisome proliferator-activated receptor γ activation ameliorates liver fibrosis-differential action of transcription factor EB and autophagy on hepatocytes and stellate cells.

Peroxisome proliferator-activated receptor γ (PPARγ) activation suppresses HSC activation and liver fibrosis. Moreover, autophagy is implicated in hepatic lipid metabolism. Here, we determined whether PPARγ activation ameliorates HSC activation by downregulating transcription factor EB (TFEB)-mediated autophagy. Atg7 or Tfeb knockdown in human HSC line LX-2 cells downregulated the expression of fibrogenic markers including α smooth muscle actin, glial fibrillary acidic protein, and collagen type 1. Conversely, Atg7 or Tfeb overexpression upregulated fibrogenic marker expression. Rosiglitazone (RGZ)-mediated PPARγ activation and/or overexpression in LX-2 cells and primary HSCs decreased autophagy, as indicated by LC3B conversion, total and nuclear-TFEB contents, mRFP-LC3 and BODIPY 493/503 colocalization, and GFP-LC3 and LysoTracker colocalization. RGZ treatment decreased liver fat content, liver enzyme levels, and fibrogenic marker expression in high-fat high-cholesterol diet-fed mice. Electron microscopy showed that RGZ treatment restored the high-fat high-cholesterol diet-mediated lipid droplet decrease and autophagic vesicle induction in primary HSCs and liver tissues. However, TFEB overexpression in LX-2 cells offset the aforementioned effects of RGZ on autophagic flux, lipid droplets, and fibrogenic marker expression. Activation of PPARγ with RGZ ameliorated liver fibrosis and downregulation of TFEB and autophagy in HSCs may be important for the antifibrotic effects of PPARγ activation.

Karakteristik Minyak Ikan Manyung (Arius sp.) dengan Metode Dry Rendering

Manyung fish (Arius sp.) is a demersal species that lives in sea and estuary waters. In Tarakan City, North Kalimantan, the meat of Manyung fish is widely consumed by the society, but the liver is discarded. One of the efforts to utilize manyung fish liver, so it does not become waste was prepare it into fish oil products. The aims of this study were to determine the yield of manyung fish (Arius sp.) liver oil, test its chemical characteristics, such as free fatty acid value (FFA), peroxide value (PV), and fatty acid profile. The method used for producing manyung fish (Arius sp.) liver oil is extraction by dry rendering at 80°C for 2 hours. The data obtained from the results of the research were then processed in a quantitative descriptive manner. The results showed that the fat content of manyung fish liver was 3.60%. The yield of manyung fish (Arius sp.) was 14.25%. The free fatty acid values (FFA) were 14.60%. The peroxide value was 3.46 mEq/kg. The fatty acid profile of mayung fish liver oil (Arius sp.) in dry rendering, the content of SFA was 0.0344%, MUFA was 0.022%, and PUFA was 0.184%. The conclusion of this research is manyung fish liver can be produced into fish oil.

Deep Learning–Based Image Analysis of Liver Steatosis in Mouse Models

The incidence of non-alcoholic fatty liver disease (NAFLD) is a continuously growing health problem worldwide, along with obesity. Therefore, both novel methods to efficiently study the manifestation of NAFLD and to analyze drug efficacy in pre-clinical models are needed. In the present study, we developed a deep neural network -based model to quantify micro- and macrovesicular steatosis in the liver on hematoxylin-eosin stained whole slide images (WSIs), using the cloud-based platform, Aiforia Create (Aiforia Technologies, Helsinki, Finland). The training data included a total of 101 WSIs from dietary interventions of wild-type mice and from two genetically modified (GM) mouse models with steatosis. The algorithm was trained for the following: to detect liver parenchyma, to exclude the blood vessels and any artefacts generated during tissue processing and image acquisition, to recognize and differentiate the areas of micro- and macrovesicular steatosis, and to quantify the recognized tissue area. The results of the image analysis replicated well the evaluation by expert pathologists, and correlated well with the liver fat content measured by EcoMRI ex vivo, and the correlation with total liver triglycerides were notable. In conclusion, the developed deep learning-based model is a novel tool for studying liver steatosis in mouse models on paraffin sections, and thus, can facilitate reliable quantification of the amount of steatosis in large preclinical study cohorts.

Bayesian networks and imaging-derived phenotypes highlight the role of fat deposition in COVID-19 hospitalisation risk.

Objective: Obesity is a significant risk factor for adverse outcomes following coronavirus infection (COVID-19). However, BMI fails to capture differences in the body fat distribution, the critical driver of metabolic health. Conventional statistical methodologies lack functionality to investigate the causality between fat distribution and disease outcomes. Methods: We applied Bayesian network (BN) modelling to explore the mechanistic link between body fat deposition and hospitalisation risk in 459 participants with COVID-19 (395 non-hospitalised and 64 hospitalised). MRI-derived measures of visceral adipose tissue (VAT), subcutaneous adipose tissue (SAT), and liver fat were included. Conditional probability queries were performed to estimate the probability of hospitalisation after fixing the value of specific network variables. Results: The probability of hospitalisation was 18% higher in people living with obesity than those with normal weight, with elevated VAT being the primary determinant of obesity-related risk. Across all BMI categories, elevated VAT and liver fat (>10%) were associated with a 39% mean increase in the probability of hospitalisation. Among those with normal weight, reducing liver fat content from >10% to <5% reduced hospitalisation risk by 29%. Conclusion: Body fat distribution is a critical determinant of COVID-19 hospitalisation risk. BN modelling and probabilistic inferences assist our understanding of the mechanistic associations between imaging-derived phenotypes and COVID-19 hospitalisation risk.

Loss of hepatic PPARα in mice causes hypertension and cardiovascular disease.

The leading cause of death in patients with nonalcoholic fatty liver disease (NAFLD) is cardiovascular disease (CVD). However, the mechanisms are unknown. Mice deficient in hepatocyte proliferator-activated receptor-α (PPARα) (PparaHepKO) exhibit hepatic steatosis on a regular chow diet, making them prone to manifesting NAFLD. We hypothesized that the PparaHepKO mice might be predisposed to poorer cardiovascular phenotypes due to increased liver fat content. Therefore, we used PparaHepKO and littermate control mice fed a regular chow diet to avoid complications with a high-fat diet, such as insulin resistance and increased adiposity. After 30 wk on a standard diet, male PparaHepKO mice exhibited elevated hepatic fat content compared with littermates as measured by Echo MRI (11.95 ± 1.4 vs. 3.74 ± 1.4%, P < 0.05), hepatic triglycerides (1.4 ± 0.10 vs. 0.3 ± 0.01 mM, P < 0.05), and Oil Red O staining, despite body weight, fasting blood glucose, and insulin levels being the same as controls. The PparaHepKO mice also displayed elevated mean arterial blood pressure (121 ± 4 vs. 108 ± 2 mmHg, P < 0.05), impaired diastolic function, cardiac remodeling, and enhanced vascular stiffness. To determine mechanisms controlling the increase in stiffness in the aorta, we used state-of-the-art PamGene technology to measure kinase activity in this tissue. Our data suggest that the loss of hepatic PPARα induces alterations in the aortas that reduce the kinase activity of tropomyosin receptor kinases and p70S6K kinase, which might contribute to the pathogenesis of NAFLD-induced CVD. These data indicate that hepatic PPARα protects the cardiovascular system through some as-of-yet undefined mechanism.

An adult-based genetic risk score for liver fat associates with liver and plasma lipid traits in children and adolescents.

Genome-wide association studies have identified steatogenic variants that also showed pleiotropic effects on cardiometabolic traits in adults. We investigated the effect of eight previously reported genome-wide significant steatogenic variants, individually and combined in a weighted genetic risk score (GRS), on liver and cardiometabolic traits, and the predictive ability of the GRS for hepatic steatosis in children and adolescents. Children and adolescents with overweight (including obesity) from an obesity clinic group (n = 1768) and a population-based group (n = 1890) were included. Cardiometabolic risk outcomes and genotypes were obtained. Liver fat was quantified using 1 H-MRS in a subset of 727 participants. Variants in PNPLA3, TM6SF2, GPAM and TRIB1 were associated with higher liver fat (p < .05) and with distinct patterns of plasma lipids. The GRS was associated with higher liver fat content, plasma concentrations of alanine transaminase (ALT), aspartate aminotransferase (AST) and favourable plasma lipid levels. The GRS was associated with higher prevalence of hepatic steatosis (defined as liver fat ≥5.0%) (odds ratio per 1-SD unit: 2.17, p = 9.7E-10). A prediction model for hepatic steatosis including GRS alone yielded an area under the curve (AUC) of 0.78 (95% CI 0.76-0.81). Combining the GRS with clinical measures (waist-to-height ratio [WHtR] SDS, ALT, and HOMA-IR) increased the AUC up to 0.86 (95% CI 0.84-0.88). The genetic predisposition for liver fat accumulation conferred risk of hepatic steatosis in children and adolescents. The liver fat GRS has potential clinical utility for risk stratification.

Alleviation of Hepatic Steatosis by Alpha-Defensin Is Associated with Enhanced Lipolysis.

Background and Objectives: The neutrophilic peptide, alpha-defensin, is considered an evolving risk factor intimately linked with lipid mobilization. It was previously linked to augmented liver fibrosis. Here, we assess a potential association between alpha-defensin and fatty liver. Materials and Methods: A cohort of transgenic C57BL/6JDef+/+ male mice that overexpress the human neutrophil-derived alpha-defensin in their polymorphonuclear neutrophils (PMNs) were assessed for liver steatosis and fibrosis development. Wild type (C57BL/6JDef.Wt) and transgenic (C57BL/6JDef+/+) mice were maintained on a standard rodent chow diet for 8.5 months. At the termination of the experiment, systemic metabolic indices and hepatic immunological cell profiling were assessed. Results: The Def+/+ transgenic mice exhibited lower body and liver weights, lower serum fasting glucose and cholesterol, and significantly lower liver fat content. These results were associated with impaired liver lymphocytes count and function (lower CD8, NK cells, and killing marker CD107a). The metabolic cage demonstrated dominant fat utilization with a comparable food intake in the Def+/+ mice. Conclusions: Chronic physiological expression of alpha-defensin induces favorable blood metabolic profile, increased systemic lipolysis, and decreased hepatic fat accumulation. Further studies are needed to characterize the defensin net liver effect.