Plasma Proteome Profiling of HbH-CS Disease Using TMT-Based Quantitative Proteomics Approach

Hemoglobin H-Constant Spring disease (HbH-CS) is the most clinically severe non-deletional form of α-thalassemia, yet its plasma proteomic characteristics remain unclear. In this study, we employed TMT-based quantitative proteomics to analyze differentially expressed plasma proteins between HbH-CS patients (n = 6) and healthy controls (n = 6), with validation performed in an independent cohort (45 cases per group) by ELISA. A total of 668 proteins were identified, among which 56 were differentially expressed (35 upregulated, 21 downregulated). Multivariate regression analysis identified NACA (OR = 1.014), HSP90AA1 (OR = 1.068), and PTGES3 (OR = 1.032) as independent risk factors (all P < 0.05). The area under the ROC curve (AUC) of NACA for diagnosing HbH-CS reached 0.911. NACA levels were negatively correlated with hemoglobin concentration (ρ=-0.397, P < 0.01), and the AUC for predicting severe anemia (Hb ≤ 80 g/L) was also 0.911. This study provides the first comprehensive plasma proteomic landscape of HbH-CS disease. NACA, as an independent risk factor, demonstrates excellent diagnostic efficacy and predictive value for anemia severity, and holds promise as a novel non-invasive biomarker for this disease.