在这里，我们介绍了能够在体外、体外和体内特异性结合胶质瘤细胞的DNA适配体，用于中枢神经系统肿瘤可视化诊断。我们使用修饰过的逐渐丰富配体的系统进化过程（SELEX）来筛选结合特异性结合于术后人类胶质原发性肿瘤而不结合于健康的脑细胞和脑膜瘤的适配体，测定了配体的序列并使用生物信息学工具分析了单链DNA池，通过其结合能力确定了最佳适配体。我们用小角X射线散射和分子建模确定了领先的适配体（Gli-55和Gli-233）的三维结构，并通过质谱法分离和鉴定了适配体的分子靶蛋白，通过分子动力学模拟验证了Gli-233与靶蛋白的潜在结合位点和后转录修饰的作用。我们利用抗胶质瘤适配体Gli-233和Gli-55检测了液体活检中的循环肿瘤细胞。这些适配体用于原位、体外组织染色、组织病理学分析以及对人类星形胶质瘤移植小鼠进行荧光引导的肿瘤和PET/CT肿瘤可视化。在预临床动物研究中，这些适配体没有显示出体内毒性。本研究展示了适配体在脑肿瘤精确诊断和荧光引导手术中的潜在应用。©2023作者。

Here, we present DNA aptamers capable of specific binding to glial tumor cells in vitro, ex vivo, and in vivo for visualization diagnostics of central nervous system tumors. We selected the aptamers binding specifically to the postoperative human glial primary tumors and not to the healthy brain cells and meningioma, using a modified process of systematic evolution of ligands by exponential enrichment to cells; sequenced and analyzed ssDNA pools using bioinformatic tools and identified the best aptamers by their binding abilities; determined three-dimensional structures of lead aptamers (Gli-55 and Gli-233) with small-angle X-ray scattering and molecular modeling; isolated and identified molecular target proteins of the aptamers by mass spectrometry; the potential binding sites of Gli-233 to the target protein and the role of post-translational modifications were verified by molecular dynamics simulations. The anti-glioma aptamers Gli-233 and Gli-55 were used to detect circulating tumor cells in liquid biopsies. These aptamers were used for in situ, ex vivo tissue staining, histopathological analyses, and fluorescence-guided tumor and PET/CT tumor visualization in mice with xenotransplanted human astrocytoma. The aptamers did not show in vivo toxicity in the preclinical animal study. This study demonstrates the potential applications of aptamers for precise diagnostics and fluorescence-guided surgery of brain tumors.© 2023 The Authors.