Circulating Extracellular Vesicles Carrying Sphingolipid Cargo for the Diagnosis and Dynamic Risk Profiling of Alcoholic Hepatitis

Abstract

Background and Aims Alcoholic hepatitis (AH) is diagnosed by clinical criteria, although several objective scores facilitate risk stratification. Extracellular vesicles (EVs) have emerged as biomarkers for many diseases and are also implicated in the pathogenesis of AH. Therefore, we investigated whether plasma EV concentration and sphingolipid cargo could serve as diagnostic biomarkers for AH and inform prognosis to permit dynamic risk profiling of AH subjects. Approach and Results EVs were isolated and quantified from plasma samples from healthy controls, heavy drinkers, and subjects with end‐stage liver disease (ESLD) attributed to cholestatic liver diseases and nonalcoholic steatohepatitis, decompensated alcohol‐associated cirrhosis (AC), and AH. Sphingolipids were quantified by tandem mass spectroscopy. The median plasma EV concentration was significantly higher in AH subjects (5.38 × 1011/mL) compared to healthy controls (4.38 × 1010/mL; P < 0.0001), heavy drinkers (1.28 × 1011/mL; P < 0.0001), ESLD (5.35 × 1010/mL; P < 0.0001), and decompensated AC (9.2 × 1010/mL; P < 0.0001) disease controls. Among AH subjects, EV concentration correlated with Model for End‐Stage Liver Disease score. When EV counts were dichotomized at the median, survival probability for AH subjects at 90 days was 63.0% in the high‐EV group and 90.0% in the low‐EV group (log‐rank P value = 0.015). Interestingly, EV sphingolipid cargo was significantly enriched in AH when compared to healthy controls, heavy drinkers, ESLD, and decompensated AC (P = 0.0001). Multiple sphingolipids demonstrated good diagnostic and prognostic performance as biomarkers for AH. Conclusions Circulating EV concentration and sphingolipid cargo signature can be used in the diagnosis and differentiation of AH from heavy drinkers, decompensated AC, and other etiologies of ESLD and predict 90‐day survival permitting dynamic risk profiling.

XIAP Knockdown in Alcohol-Associated Liver Disease Models Exhibits Divergent in vitro and in vivo Phenotypes Owing to a Potential Zonal Inhibitory Role of SMAC

“…Cells were cultured in an EV free 10% FBS medium. EVs were then isolated from cultured cells or human plasma samples as described earlier (Sehrawat et al, 2020a). Supernatants from cultured cells were subjected to differential ultracentrifugation for 45 mins (20,000 g) and twice for 2 hrs (110,000 g) at 4 • C. Determination of EV concentration and size was completed using nanoparticle tracking analysis (NTA) (NS300, Malvern Instruments, Malvern, United Kingdom).…”

Extracellular Vesicles in NAFLD/ALD: From Pathobiology to Therapy

“…Additionally, a very recent study showed that circulating EVs carrying a sphingolipid cargo might be helpful for the diagnosis of AH, and may allow dynamic risk profiling of these patients. [98]. Most recently, EVs have been used as a surrogate marker of improvement in clinical trials of patients with AH [99].…”

Roles of Macrophages and Exosomes in Liver Diseases

“…Exosomes can also be used for prognostic analyses. Circulating EV concentrations and sphingolipid carrier characteristics can be used not only for the diagnosis and differentiation of AH, decompensated alcoholic cirrhosis, and other end-stage liver diseases but also for the prediction of the 90-day survival time ( 101 ).…”