Serban Georgiana M, Doina Manu, Balasa Rodica, Balasa Adrian F
Doctoral School, "George Emil Palade" University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Targu Mures, ROU.
Anesthesiology and Critical Care, Emergency Clinical County Hospital of Targu Mures, Targu Mures, ROU.
Cureus. 2024 Sep 28;16(9):e70403. doi: 10.7759/cureus.70403. eCollection 2024 Sep.
Glioblastoma (GB) is the most common brain malignancy occurring in adult patients having an extremely low overall survival. Therefore, it is paramount to establish reliable and accurate diagnostic and prognostic markers to guide a personalized and more effective treatment. Molecular characterization of the tumor is the ultimate goal in GB management and comprises, among others, the study of the extracellular vesicles (EVs). Not only do they carry within their cargo molecules involved in shaping a favorable microenvironment for GB development, but EVs also present surface markers mirroring the phenotype of the donor cells. Our study aims to assess the dynamic evolution of EV-positive surface biomarkers and EV-derived proteins involved in maintaining and transferring a stem cell phenotype to the cells from GB surroundings. We performed a prospective observational study on GB patients operated on in the Neurosurgery Clinic of the Emergency Clinical County Hospital of Târgu Mureș, Romania. GB-derived EVs were isolated from the patients' plasma using a density gradient ultracentrifugation protocol. The expression of EVs positive to four epitopes specific to stem cells (CD44, CD133, CD326/EpCAM, and SSEA4) was followed in three moments in time, preoperatively, seven days, and three months postoperatively, respectively, and quantified by a bead-based multiple analysis using flow cytometry. Moreover, NANOGP8, a protein within GB cargo capable of promoting a stem cell phenotype, was dynamically evaluated using the Western blot technique. Our study showed a statistically significant decrease of all surface markers and NANOGP8 immediately after tumor ablation. Nonetheless, the long-term follow-up of the patients revealed an extremely variable evolutionary pattern reflecting the high heterogeneity of GB. Further studies are necessary to either confirm or infirm the accuracy of these markers in early diagnosing GB, in predicting the outcome of this disease, and in guiding an individualized therapy.
胶质母细胞瘤（GB）是成年患者中最常见的脑恶性肿瘤，总体生存率极低。因此，建立可靠且准确的诊断和预后标志物以指导个性化且更有效的治疗至关重要。肿瘤的分子特征是胶质母细胞瘤治疗的最终目标，其中包括对细胞外囊泡（EVs）的研究。它们不仅在其货物分子中携带参与为胶质母细胞瘤发展塑造有利微环境的分子，而且细胞外囊泡还呈现反映供体细胞表型的表面标志物。我们的研究旨在评估参与维持干细胞表型并将其转移至胶质母细胞瘤周围细胞的细胞外囊泡阳性表面生物标志物和细胞外囊泡衍生蛋白的动态演变。我们对在罗马尼亚特尔古穆列什市急诊临床县医院神经外科诊所接受手术的胶质母细胞瘤患者进行了一项前瞻性观察研究。使用密度梯度超速离心方案从患者血浆中分离出胶质母细胞瘤衍生的细胞外囊泡。分别在术前、术后七天和术后三个月这三个时间点跟踪对四种干细胞特异性表位（CD44、CD133、CD326/EpCAM和SSEA4）呈阳性的细胞外囊泡的表达，并通过基于微珠的流式细胞术多重分析进行定量。此外，使用蛋白质免疫印迹技术动态评估了胶质母细胞瘤货物中能够促进干细胞表型的一种蛋白质NANOGP8。我们的研究表明，肿瘤切除后所有表面标志物和NANOGP8立即出现统计学上的显著下降。尽管如此，对患者的长期随访显示出一种极其多变的演变模式，反映了胶质母细胞瘤的高度异质性。需要进一步研究以确认或否定这些标志物在早期诊断胶质母细胞瘤、预测该疾病的预后以及指导个体化治疗方面的准确性。
