Background:Oncological diseases are a major focus in medicine, with millions diagnosed each year, leading researchers to seek new diagnostic and treatment methods. One promising avenue is the development of targeted therapies and rapid diagnostic tests using recognition molecules. The pharmaceutical industry is increasingly exploring nucleic acid-based therapeutics. However, producing long oligonucleotides, especially aptamers, poses significant production challenges.Objectives:This study aims to demonstrate the efficacy of using molecular modeling, supported by experimental procedures, for altering aptamer nucleotide sequences while maintaining their binding capabilities. The focus is on reducing production costs and enhancing binding dynamics by removing nonfunctional regions and minimizing nonspecific binding.Methods:A molecular modeling approach was employed to elucidate the structure of a DNA aptamer, Gli-55, facilitating the truncation of nonessential regions in the Gli-55 aptamer, which selectively binds to glioblastoma (GBM). This process aimed to produce a truncated aptamer, Gli-35, capable of forming similar structural elements to the original sequence with reduced nonspecific binding. The efficiency of the truncation was proved by flow cytometry, fluorescence polarization (FP), and confocal microscopy.Results:The molecular design indicated that the new truncated Gli-35 aptamer retained the structural integrity of Gli-55. In vitro studies showed that Gli-35 had a binding affinity comparable to the initial long aptamer while the selectivity increased. Gli-35 internalized inside the cell faster than Gli-55 and crossed the blood–brain barrier (BBB), as demonstrated in an in vitro model.Conclusions:The success of this truncation approach suggests its potential applicability in scenarios where molecular target information is limited. The study highlights a strategic and resource-efficient methodology for aptamer development. By employing molecular modeling and truncation, researchers can reduce production costs and avoid trial and error in sequence selection. This approach is promising for enhancing the efficiency of therapeutic agent development, particularly in cases lacking detailed molecular target insights.
背景：肿瘤疾病是医学领域的重要焦点，每年有数百万人被诊断，促使研究人员寻求新的诊断和治疗方法。其中，利用识别分子开发靶向疗法和快速诊断测试是一条前景广阔的途径。制药行业正日益探索基于核酸的治疗方法。然而，生产长链寡核苷酸，特别是适配体，面临着显著的生产挑战。 目的：本研究旨在证明，在实验程序的支持下，利用分子建模改变适配体核苷酸序列同时保持其结合能力的有效性。重点是通过去除非功能区域和减少非特异性结合来降低生产成本并增强结合动力学。 方法：采用分子建模方法阐明DNA适配体Gli-55的结构，促进对选择性结合胶质母细胞瘤（GBM）的Gli-55适配体中非必需区域的截短。此过程旨在产生截短的适配体Gli-35，其能够形成与原始序列相似的结构元件，同时减少非特异性结合。通过流式细胞术、荧光偏振（FP）和共聚焦显微镜证明了截短的有效性。 结果：分子设计表明，新的截短适配体Gli-35保持了Gli-55的结构完整性。体外研究显示，Gli-35的结合亲和力与初始长链适配体相当，同时选择性有所提高。在体外模型中证明，Gli-35比Gli-55更快地内化到细胞内部，并能穿过血脑屏障（BBB）。 结论：这种截短方法的成功表明其在分子靶标信息有限的情况下具有潜在适用性。该研究强调了一种战略性和资源高效的适配体开发方法。通过采用分子建模和截短技术，研究人员可以降低生产成本，并避免序列选择中的试错过程。这种方法有望提高治疗剂开发的效率，特别是在缺乏详细分子靶标信息的情况下。
Aptamer’s Structure Optimization for Better Diagnosis and Treatment of Glial Tumors
肿瘤(癌症)患者之家