Prospective Biopsy-Controlled Evaluation of Cell Death Biomarkers for Prediction of Liver Fibrosis and Nonalcoholic Steatohepatitis

Abstract

Fibrosis and steatosis are major histopathological alterations in chronic liver diseases. Despite various shortcomings, disease severity is generally determined by liver biopsy, emphasizing the need for simple noninvasive methods for assessing disease activity. Because hepatocyte cell death is considered a crucial pathogenic factor, we prospectively evaluated the utility of serum biomarkers of cell death to predict different stages of fibrosis and steatosis in 121 patients with chronic liver disease. We compared the M30 enzyme‐linked immunosorbent assay (ELISA), which detects a caspase‐cleaved cytokeratin‐18 (CK‐18) fragment and thereby apoptotic cell death, with the M65 ELISA, which detects both caspase‐cleaved and uncleaved CK‐18 and thereby overall cell death. Both biomarkers significantly discriminated patients with different fibrosis stages from healthy controls. However, whereas both markers differentiated low or moderate from advanced fibrosis, only the M65 antigen could discriminate even lower stages of fibrosis. The M65 assay also performed better in distinguishing low (≤10%) and higher (>10%) grades of steatosis. In a subgroup of patients, we evaluated the biomarkers for their power to predict nonalcoholic steatohepatitis (NASH). Importantly, both markers accurately differentiated healthy controls or simple steatosis from NASH. However, only serum levels of M65 antigen could differentiate simple steatosis from healthy controls. Conclusion: Cell death biomarkers are potentially useful to predict fibrosis, steatosis, or NASH. Compared with the widely used apoptosis marker M30, the M65 assay had a better diagnostic performance and even differentiated between lower fibrosis stages as well as between healthy individuals and patients with simple steatosis. (HEPATOLOGY 2012)

Assessment of non‐alcoholic fatty liver disease using serum total cell death and apoptosis markers

“…However, we chose the assays that were used in recent similar studies to facilitate comparison across studies. 9,24 In conclusion, apoptosis marker CK-18 M30 and total cell death marker M65/M65ED have similar high diagnostic performance in NAFLD and similar moderate accuracy in predicting NASH. Changes in the biomarkers also correlate with histological progression.…”

“…21,22 However, Joka and colleagues suggested recently that necrosis might also contribute to the liver damage in 5000 4000 3000 2000 NAFLD and NASH by showing biomarkers detecting both apoptosis and necrosis might be superior to detecting apoptosis alone. 24 However, this study included patients with different liver diseases, and NAFLD patients were underrepresented. The role of different cell death markers was also discussed in other liver diseases, such as HBV infection and acute liver failure.…”

“…23 Joka and colleagues suggested that M65/ M65ED may have superior performance to M30 in detecting mild fibrosis and steatosis. 24 However, that study was limited by the inclusion of different liver diseases and the small number of NAFLD patients. 25 As such, the performance of M65 and M65ED in detecting NASH compared with M30 is currently unknown.…”

Association of Serum Cytokeratin-18 Fragment Concentration in Patients with Different Types of Nonalcoholic Fatty Liver Disease

“…CK-18 fragment level did not differ significantly between the patients with simple fatty liver and borderline NASH also. Joka et al [12] also observed that, CK-18 fragment could make discrimination between simple fatty liver and NASH. In the study of Wieckowska et al [13] CK-18 fragment levels were significantly increased in the blood of patients with NASH compared with patients with simple steatosis.…”

“…Tsutsui et al [15] also observed that there was no significant correlation between fibrosis and CK-18 fragment level. On the other hand, Joka et al [12] observed a positive significant correlation between CK-18 fragment level and fibrosis score12. However, our study finding was not consistent with the studies conducted by Joka et al [12].…”