Evaluation of Zonulin Expression and Its Potential Clinical Significance in Glioblastoma

Simple Summary Glioblastoma, the most deadly type of brain tumor in adults, poses a significant treatment challenge and often leads to poor outcomes. A recent study delved into the role of a protein called zonulin, known for its impact on the body's barrier functions, in glioblastoma. The research involved examining zonulin levels in both the bloodstream and tumor tissues of 34 newly diagnosed glioblastoma patients. The study discovered that higher levels of zonulin in tumors were linked to increased aggressiveness, which in turn correlated with a poorer prognosis. Elevated serum levels of zonulin in the blood also indicated a connection with a less favorable prognosis. In laboratory experiments, glioblastoma cells expressed higher levels of zonulin, particularly under conditions resembling glioma stem cells, which are associated with treatment resistance. This research suggests that heightened zonulin levels in both tumors and the blood may signal a more challenging prognosis for glioblastoma patients, underscoring the potential importance of this protein in the disease.Abstract Glioblastoma, the deadliest adult brain tumor, poses a significant therapeutic challenge with a dismal prognosis despite current treatments. Zonulin, a protein influencing tight junctions and barrier functions, has gained attention for its diverse roles in various diseases. This study aimed to preliminarily analyze the circulating and tumor zonulin levels, evaluating their impact on disease prognosis and clinical-radiological factors. Additionally, we investigated in vitro zonulin expression in different glioblastoma cell lines under two different conditions. The study comprised 34 newly diagnosed glioblastoma patients, with blood samples collected before treatment for zonulin and haptoglobin analysis. Tumor tissue samples from 21 patients were obtained for zonulin expression. Clinical, molecular, and radiological data were collected, and zonulin protein levels were assessed using ELISA and Western blot techniques. Furthermore, zonulin expression was analyzed in vitro in three glioblastoma cell lines cultured under standard and glioma-stem-cell (GSC)-specific conditions. High zonulin expression in glioblastoma tumors correlated with larger preoperative contrast enhancement and edema volumes. Patients with high zonulin levels showed a poorer prognosis (progression-free survival [PFS]). Similarly, elevated serum levels of zonulin were associated with a trend of shorter PFS. Higher haptoglobin levels correlated with MGMT methylation and longer PFS. In vitro, glioblastoma cell lines expressed zonulin under standard cell culture conditions, with increased expression in tumorsphere-specific conditions. Elevated zonulin levels in both the tumor and serum of glioblastoma patients were linked to a poorer prognosis and radiological signs of increased disruption of the blood-brain barrier. In vitro, zonulin expression exhibited a significant increase in tumorspheres.