P33 Sera metabolite changes in obesity-induced MASLD, assayed with metabolomic approaches

<h3>Background</h3> Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD), formerly known as Non-alcoholic fatty liver disease (NAFLD) is one of the leading causes of chronic liver disease, affecting 25% globally.¹ It starts as hepatic steatosis, progresses to steatohepatitis, fibrosis, cirrhosis, and possibly hepatocellular cancer.² MASLD is often associated with obesity and Type 2 diabetes.^3–7 Liver function blood tests &amp; non-invasive radiological imaging are clinically used to detect MASLD. Whilst, liver biopsy remains the ‘gold standard’ for detection &amp; staging, the procedure is invasive &amp; can be unreliable especially in obese patients.⁸ There is a need for specific &amp; non-invasive diagnostic biomarkers for the early detection &amp; staging of MASLD. Our aim was to assay sera metabolite changes, as our previous investigation showed urinary metabolome changes in mice with obesity-induced MASLD. <h3>Methods</h3> C57BL/6 mice (n=78) were fed a diet comprising high-fat(54%), cholesterol(1.4%) plus sucrose(55%) &amp; fructose(45%) in their drinking water, to simulate the diet typically consumed by obese patients with MASLD. Sera of mice fed the junk diet (JC) with their parallel controls fed normal chow (NC) for 6,8 &amp; 10months, were assayed using metabolomic techniques. These techniques were Fourier-transform infrared (FT-IR), 1H nuclear magnetic resonance (NMR), 2D 1H–1H total correlation (TOCSY), heteronuclear single quantum coherence &amp; electrospray liquid chromatography-mass spectrometry (ESI-LC-MS/MS) with multivariate analysis. Body, liver &amp; liver/body weights, liver biochemistry tests &amp; histology were evaluated. <h3>Results and Conclusion</h3> Sera ALT were elevated in JC groups compared with normal chow, NC (57.57±10.84 IU/L vs 501.60±54.48IU/L; 97.00±24.82 IU/L vs 613.30±56.69 IU/L; 101.70±15.79 IU/L vs 401.40±10.57 IU/L for the 6,8 &amp; 10months, respectively, compared to NC, all p&lt;0.01). Sera AST levels were increased in JC cohorts (201.70±29.28 U/L vs 541.10±54.97 U/L; 246.60±42.86 U/L vs 665.60±66.12 U/L; 367.00±45.58 U/L vs 544.00±21.75 U/L for the 6,8 &amp; 10months, respectively, all p&lt;0.01). The MASLD clinical research network score for the JC livers increased (0.14 vs 4.8; 0.6 vs 6.0; 1.0 vs 5.1 for the 6,8 &amp; 10months, respectively, all p&lt;0.001) compared with NC. Principal component analysis scores plot (figure 1) sera FT-IR spectra displayed differences between the NC and JC groups, indicating that JC induced biomolecule level changes. We are now identifying the sera metabolites with 1H NMR, 2D 1H–1H TOCSY and ESI-LC-MS/MS and planning parallel human studies. The results demonstrate that metabolomic approaches may be useful in detecting novel biomarkers. <h3>References</h3> Huang DQ, HB El-Serag, R Loomba. Global epidemiology of MASLD-related HCC: trends, predictions, risk factors and prevention. <i>Nat Rev Gastroenterol Hepatol</i>, 2021;<b>18</b>(4):223–238. Ekstedt M, <i>et al</i>. Long-term follow-up of patients with MASLD and elevated liver enzymes. <i>Hepatology</i>, 2006;<b>44</b>(4):865–73. Amacher DE, N Chalasani. Drug-induced hepatic steatosis. <i>Semin Liver Dis</i>, 2014;<b>34</b>(2):205–14.