Neves Ana R, Vivès Eric, Boisguérin Prisca, Quintela Telma, Costa Diana
RISE-Health, Department of Medical Sciences, Faculty of Health Sciences, University of Beira Interior, Av. Infante D. Henrique, 6200-506 Covilhã, Portugal.
PhyMedExp, University of Montpellier, INSERM, CNRS, 34295 Montpellier, France.
Int J Mol Sci. 2025 Jun 26;26(13):6130. doi: 10.3390/ijms26136130.
Glioblastoma is the most prevalent and aggressive form of brain malignancy. Actual treatments face several challenges due to its high aggressiveness and poor prognosis. The chemotherapeutic agent temozolomide (TMZ) has limited therapeutic efficacy, and mutations in the tumour protein p53 gene () have been associated with treatment resistance. Thus, this study aimed to explore an innovative therapeutic strategy to enhance treatment efficacy of GBM. Previously, our team had developed a WRAP5 cell-penetrating peptide (CPP) functionalized with a transferrin receptor ligand (Tf) for the targeted delivery of TMZ and a p53-encoding plasmid to glioma cells. Our research had elucidated the circadian oscillations of the clock genes in the U87 glioma cells by employing two different computational models and observed that T16 and T8 time points revealed the highest circadian activity for and genes, respectively. Similar analysis was conducted for the transferrin receptor, which revealed that T7 and T8 were the key time points for its expression. A confocal microscopy study indicated the highest intracellular uptake of complexes and p53 mRNA expression at T8, the time point with the highest Per2 and transferrin receptor expression. Following mRNA analysis, the evaluation of p53 levels confirmed transcriptional changes at the protein level, and that T16 appears to be a favourable time point for enhancing therapeutic efficacy in U87 glioblastoma cells. These findings suggested that synchronizing the complexes' administration with the biological clock of GBM cells may significantly improve glioblastoma therapeutics.
胶质母细胞瘤是最常见且侵袭性最强的脑恶性肿瘤形式。由于其高度侵袭性和不良预后，目前的治疗面临诸多挑战。化疗药物替莫唑胺（TMZ）的治疗效果有限，肿瘤蛋白p53基因（）的突变与治疗耐药性相关。因此，本研究旨在探索一种创新的治疗策略，以提高胶质母细胞瘤的治疗效果。此前，我们的团队开发了一种用转铁蛋白受体配体（Tf）功能化的WRAP5细胞穿透肽（CPP），用于将TMZ和p53编码质粒靶向递送至胶质瘤细胞。我们的研究通过采用两种不同的计算模型阐明了U87胶质瘤细胞中生物钟基因的昼夜节律振荡，并观察到T16和T8时间点分别显示出和基因的最高昼夜活性。对转铁蛋白受体进行了类似分析，结果表明T7和T8是其表达的关键时间点。共聚焦显微镜研究表明，在T8（Per2和转铁蛋白受体表达最高的时间点），复合物的细胞内摄取和p53 mRNA表达最高。mRNA分析后，对p53水平的评估证实了蛋白质水平的转录变化，并且T16似乎是提高U87胶质母细胞瘤细胞治疗效果的有利时间点。这些发现表明，使复合物的给药与胶质母细胞瘤细胞的生物钟同步可能会显著改善胶质母细胞瘤的治疗效果。
