iTRAQ-based Quantitative Proteomic Analysis of Dural Tissues Reveals Upregulated Haptoglobin to be a Potential Biomarker of Moyamoya Disease

Background: Moyamoya Disease (MMD) is a rare cerebrovascular disease with a high rate of disability and mortality. Immune reactions have been implicated in the pathogenesis of MMD, however, the underlying mechanism is still unclear. Objective: To identify proteins related to MMD specially involved in the immunogenesis, we performed a proteomic study. Methods: In this work, dural tissues or plasma from 98 patients with MMD, 17 disease controls without MMD, and 12 healthy donors were included. Proteomic profiles of dural tissues from 4 MMD and 4 disease controls were analyzed by an isobaric tag for relative and absolute quantitation (iTRAQ)- based proteomics. The immune-related proteins were explored by bioinformatics and the key MMDrelated proteins were verified by western blot, multiple reaction monitoring methods, enzyme-linked immunosorbent assay, and tissue microarray. Results: 1,120 proteins were identified, and 82 MMD-related proteins were found with more than 1.5 fold difference compared with those in the control samples. Gene Ontology analysis showed that 29 proteins were immune-related. In particular, Haptoglobin (HP) was up-regulated in dural tissue and plasma of MMD samples compared to the controls, and its up-regulation was found to be sex- and MMD Suzuki grade dependent. Through Receiver Operating Characteristic (ROC) analysis, HP can well discriminate MMD and healthy donors with the Area Under the Curve (AUC) of 0.953. Conclusion: We identified the biggest protein database of the dura mater. 29 out of 82 differentially expressed proteins in MMD are involved in the immune process. Of which, HP was up-regulated in dural tissue and plasma of MMD, with sex- and MMD Suzuki grade-dependence. HP might be a potential biomarker of MMD.