神经胶质瘤是最常见的恶性脑肿瘤类型,由神经胶质细胞产生。它们约占所有原发性脑肿瘤的 40%,约占所有原发性恶性脑肿瘤的 70%。它们可发生在中枢神经系统 (CNS) 的任何部位,且预后较差。神经胶质瘤患者的平均生存期约为 6-15 个月,生活质量较差。在这一领域,对癌细胞分泌的蛋白质的鉴定特别令人感兴趣,因为它可以更好地了解肿瘤进展并提供疾病的早期诊断。从健康对照 (n=03) 和不同级别神经胶质瘤患者 (I 级、II 级或 III 级,各 n=03) 的合并血浆中分离出细胞外囊泡 (EV)。进行纳米颗粒跟踪分析、蛋白质印迹和流式细胞术以确定胶质瘤来源的囊泡的大小、形态、浓度和 EV 标记 CD63。此外,对 EV 蛋白进行基于 iTRAQ 的 LC-MS/MS 分析,以确定不同神经胶质瘤级别的细胞外囊泡中的差异蛋白丰度。我们通过 ELISA 进一步验证了对照和神经胶质瘤患者(各 n=40)的个体血浆和血浆源性囊泡中的半乳糖凝集素 3 结合蛋白(LGALS3BP)。Max Quant 的分析从合并的患者外泌体中鉴定出了 123 种蛋白质,其中 34、21 和 14 种蛋白质被发现在不同级别的胶质瘤 I 级毛细胞星形细胞瘤中丰度差异超过 1.3 倍;II级,弥漫性星形细胞瘤;与对照样本相比,分别为III级,间变性星形细胞瘤。总共七种蛋白质——即 CRP、SAA2、SERPINA3、SAA1、C4A、LV211 和 KV112——在所有三个等级中都显示出不同的丰度。LGALS3BP 在不同级别中均上调,并且对个体血浆和血浆来源的细胞外囊泡的 ELISA 分析证实了神经胶质瘤患者中 LGALS3BP 的表达增加(p<0.001)。本研究提供 LGALS3BP 作为早期检测神经胶质瘤并改善患者生存结果的潜在生物标志物。本研究进一步提供进展信息和监测肿瘤等级(1级、II级、III级)。
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Plasma-Derived Extracellular Vesicles Reveal Galectin-3 Binding Protein as Potential Biomarker for Early Detection of Glioma
Gliomas are the most common type of the malignant brain tumor, which arise from glial cells. They make up about 40% of all primary brain tumors and around 70% of all primary malignant brain tumors. They can occur anywhere in the central nervous system (CNS) and have a poor prognosis. The average survival of glioma patients is approximately 6–15 months with poor aspects of life. In this edge, identification of proteins secreted by cancer cells is of special interest because it may provide a better understanding of tumor progression and provide early diagnosis of the diseases. Extracellular vesicles (EVs) were isolated from pooled plasma of healthy controls (n=03) and patients with different grades of glioma (Grade I or II or III, n=03 each). Nanoparticle tracking analysis, western blot, and flow cytometry were performed to determine the size, morphology, the concentration of glioma-derived vesicles and EV marker, CD63. Further, iTRAQ-based LC-MS/MS analysis of EV protein was performed to determine the differential protein abundance in extracellular vesicles across different glioma grades. We further verified galectin-3 binding protein (LGALS3BP) by ELISA in individual blood plasma and plasma-derived vesicles from control and glioma patients (n=40 each). Analysis by Max Quant identified 123 proteins from the pooled patient exosomes, out of which 34, 21, and 14 proteins were found to be differentially abundant by more than 1.3-fold in the different grades of glioma grade I, pilocytic astrocytoma; grade II, diffuse astrocytoma; grade III, anaplastic astrocytoma, respectively, in comparison with the control samples. A total of seven proteins—namely, CRP, SAA2, SERPINA3, SAA1, C4A, LV211, and KV112—showed differential abundance in all the three grades. LGALS3BP was seen to be upregulated across the different grades, and ELISA analysis from individual blood plasma and plasma-derived extracellular vesicles confirmed the increased expression of LGALS3BP in glioma patients (p<0.001). The present study provides LGALS3BP as a potential biomarker for early detection of glioma and improve survival outcome of the patient. The present study further provides the information of progression and monitoring the tumor grades (grade 1, grade II, grade III).