biostudies-literatureVadivalagan CNCI NIH HHSDivision of Cancer Prevention, National Cancer Institute6079-6090https://www.ebi.ac.uk/biostudies/studies/S-EPMC12701774biostudies-literatureUnknown24(12)Hepatocellular carcinoma (HCC), commonly associated with cirrhosis, is a major cause of cancer-related mortality due to its poor prognosis. Herein, we investigated fucosylated glycoforms of serum haptoglobin (Hp) as potential biomarkers for HCC of metabolic associated dysfunction associated liver disease (MASLD) and alcohol related liver disease (ALD) etiologies. We analyzed 119 patient samples, including 60 with cirrhosis and 59 with HCC. Isolated Hp protein was digested using trypsin and Glu-C, and site-specific <i>N</i>-glycans were quantified using PRM with LC-HCD-MS/MS. Differential analysis revealed significant variations in fucosylated tetra-antennary glycoforms at N241(VVLHPN²⁴¹YSQVD and VVLHPN²⁴¹YSQVDIGLIK), particularly in distinguishing cirrhosis and HCC (<i>P</i> < 0.05). A combined analysis of identified tetra-antennary fucosylated markers, along with AFP, gender, and age, demonstrated improved AUC. Tetra-antennary glycoforms exhibited an AUC of 0.871 (95% CI: 0.80-0.93) when incorporated into the AFP + age + gender + marker panel compared to AFP alone (0.756) with a sensitivity of 0.763 at a specificity of 0.80. 3-fold cross validation was further used to assess the performance of the optimal biomarker panel. Thus, a combination of fucosylated tetra-antennary glycoforms may serve as important markers for distinguishing HCC from cirrhosis.Journal of proteome researchIntegrative Analysis of Fucosylated Tetra Glycoforms in Hepatocellular Carcinoma: A NanoLC-PRM-MS/MS and Machine Learning Approach.PMC12701774R01 CA160254R01-CA160254-11U01 CA283935U01 CA271887CA283935Vadivalagan CZhang JBass KSingal AGParikh NDLiu SDai JLubman DMfalseIntegrative Analysis of Fucosylated Tetra Glycoforms in Hepatocellular Carcinoma: A NanoLC-PRM-MS/MS and Machine Learning Approach.Hepatocellular carcinoma (HCC), commonly associated with cirrhosis, is a major cause of cancer-related mortality due to its poor prognosis. Herein, we investigated fucosylated glycoforms of serum haptoglobin (Hp) as potential biomarkers for HCC of metabolic associated dysfunction associated liver disease (MASLD) and alcohol related liver disease (ALD) etiologies. We analyzed 119 patient samples, including 60 with cirrhosis and 59 with HCC. Isolated Hp protein was digested using trypsin and Glu-C, and site-specific <i>N</i>-glycans were quantified using PRM with LC-HCD-MS/MS. Differential analysis revealed significant variations in fucosylated tetra-antennary glycoforms at N241(VVLHPN²⁴¹YSQVD and VVLHPN²⁴¹YSQVDIGLIK), particularly in distinguishing cirrhosis and HCC (<i>P</i> < 0.05). A combined analysis of identified tetra-antennary fucosylated markers, along with AFP, gender, and age, demonstrated improved AUC. Tetra-antennary glycoforms exhibited an AUC of 0.871 (95% CI: 0.80-0.93) when incorporated into the AFP + age + gender + marker panel compared to AFP alone (0.756) with a sensitivity of 0.763 at a specificity of 0.80. 3-fold cross validation was further used to assess the performance of the optimal biomarker panel. Thus, a combination of fucosylated tetra-antennary glycoforms may serve as important markers for distinguishing HCC from cirrhosis.2025-01-01T00:00:00Z2025 Dec2026-06-06T03:39:24.239Z2026-05-25T03:07:47.298ZS-EPMC127017744121202710.1021/acs.jproteome.5c00626