Donofrio Carmine Antonio, Fioravanti Antonio, Riccio Lucia, Cominetti Marika, Cappelletti Maria Rosa, Longo Giuseppe, Varotti Elena, Generali Daniele, Servadei Franco, Grossi Ilaria, Pelisenco Iulia Andreea, Salvi Alessandro, De Petro Giuseppina
Department of Neurosurgery, ASST Cremona, Viale Concordia 1, Cremona, 26100, Italy.
Division of Biology and Genetics, Department of Molecular and Translational Medicine, University of Brescia, Viale Europa 11, Brescia, 25123, Italy.
Neurosurg Rev. 2025 Aug 7;48(1):592. doi: 10.1007/s10143-025-03747-z.
Glioblastoma (GBM) is characterized by molecular heterogeneity and rapid progression. Liquid biopsy is increasingly recognised as a promising non-invasive biomarker for GBM diagnosis, prognosis and treatment monitoring. The aim of the study was to explore circulating microRNAs (miRNA) as potential GBM-biomarkers. We developed droplet digital PCR (ddPCR) assays to quantify plasmatic concentrations of miR-21-5p, miR-23b-3p and miR-34a-5p both in healthy volunteers and GBM patients at diagnosis and over follow-up (1, 3, 6 and 12 months after surgery). Furthermore, we investigated the correlations between peripheral miRNA levels and clinical, neuroradiological, pathological features, extent of tumour resection, overall (OS) and recurrence-free (RFS) survivals. Our findings showed that these miRNAs were detectable in all samples, even if with different profiles, showing a similar longitudinal course characterized by a gradual increase 1 and 3 months after surgery, followed by a progressive decrease 6 and 12 months after surgery. miR-34a-5p levels were significantly higher in GBM patients compared to healthy volunteers (AUC = 0.664, p = 0.039; cut-off: 1.25 copies/µL). RFS (7.6 vs. 15.6 months, p = 0.049) and OS (8.2 vs. 24.5 months, p = 0.032) were significantly shorter in patients with miR-34a-5p levels below the mean at diagnosis and 3 months after surgery, respectively. Similarly, OS (13.3 vs. 24.4months, p = 0.024) was significantly shorter in patients with miR-21-5p levels below the mean 6 months after surgery. This study highlights the potential clinical utility of ddPCR-based quantification of plasmatic miRNAs in GBM. Longitudinal analysis revealed consistent dynamic expression patterns for all three investigated miRNAs, with miR-34a-5p and miR-21-5p emerging as potential prognostic biomarkers. Although the diagnostic performance of miR-34a-5p was intermediate and the small cohort size limited definitive conclusions, these preliminary findings support further exploration of these miRNAs as part of a multi-marker panel to enhance diagnostic and prognostic accuracy in GBM. Larger, prospective studies are required to validate these results and to elucidate the biological underpinnings of peripheral miRNA dynamics in the context of GBM pathophysiology and treatment.
胶质母细胞瘤（GBM）具有分子异质性和快速进展的特点。液体活检越来越被认为是一种有前景的用于GBM诊断、预后评估和治疗监测的非侵入性生物标志物。本研究的目的是探索循环微小RNA（miRNA）作为潜在的GBM生物标志物。我们开发了液滴数字PCR（ddPCR）检测方法，以定量健康志愿者以及GBM患者在诊断时和随访期间（术后1、3、6和12个月）血浆中miR-21-5p、miR-23b-3p和miR-34a-5p的浓度。此外，我们研究了外周血miRNA水平与临床、神经放射学、病理特征、肿瘤切除范围、总生存期（OS）和无复发生存期（RFS）之间的相关性。我们的研究结果表明，这些miRNA在所有样本中均可检测到，即使其表达谱不同，呈现出相似的纵向变化过程，其特征为术后1个月和3个月逐渐升高，随后在术后6个月和12个月逐渐下降。与健康志愿者相比，GBM患者中miR-34a-5p水平显著更高（AUC = 0.664，p = 0.039；临界值：1.25拷贝/μL）。诊断时和术后3个月miR-34a-5p水平低于均值的患者，其RFS（7.6个月对15.6个月，p = 0.049）和OS（8.2个月对24.5个月，p = 0.032）显著更短。同样，术后6个月miR-21-5p水平低于均值的患者，其OS（13.3个月对24.4个月，p = 0.024）显著更短。本研究强调了基于ddPCR定量血浆miRNA在GBM中的潜在临床应用价值。纵向分析显示，所有三种研究的miRNA都具有一致的动态表达模式，其中miR-34a-5p和miR-21-5p成为潜在的预后生物标志物。尽管miR-34a-5p的诊断性能中等，且样本量较小限制了得出确定性结论，但这些初步发现支持进一步探索这些miRNA，作为多标志物组合的一部分，以提高GBM的诊断和预后准确性。需要更大规模的前瞻性研究来验证这些结果，并阐明在GBM病理生理学和治疗背景下外周血miRNA动态变化的生物学基础。
