Rapid identification of specific DNA aptamers precisely targeting CD33 positive leukemia cells through a paired cell-based approach.
Rapid identification of specific DNA aptamers precisely targeting CD33 positive leukemia cells through a paired cell-based approach.
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DOI：
10.1039/c8bm01393d
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发表时间：
2019-02
影响因子：
6.6
通讯作者：
Chang Yang;Yun Wang;M. Ge;Yushi Fu;Rui Hao;Khairul Islam;Ping Huang;Fu Chen;Jie Sun
中科院分区：
文献类型：
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作者：
Chang Yang;Yun Wang;M. Ge;Yushi Fu;Rui Hao;Khairul Islam;Ping Huang;Fu Chen;Jie Sun
核酸适体是一种短的单链DNA或RNA分子，近年来被开发用于临床治疗、诊断和肿瘤靶向给药等方面。然而，使用经典的蛋白质或基于细胞的SELEX来选择特异性适体通常是不令人满意的。在此，我们修改了配对细胞系方法以鉴定靶向表达CD 33抗原的白血病细胞的适体。我们的策略巧妙地使用相同的细胞进行阴性（HEK 293 T细胞）和阳性（CD 33转染的HEK 293 T细胞）适体选择，并且在阳性选择之前充分进行阴性选择以去除非特异性序列。该策略的优点是快速且准确，其中仅几轮选择连同PCR扩增就足以获得高结合亲和力的靶向抗原的适体。利用我们改进的方法，我们成功地获得了靶向CD 33的适配子S30，它在体外和体内都能高度识别CD 33抗原的C2结构域。此外，优化的适体S30-T1（即，将S30的核心区与阿霉素（Dox）偶联，合成S30-T1-Dox偶联物，该偶联物可特异性抑制CD 33阳性急性髓系白血病HL-60细胞的增殖，使细胞周期阻滞于G2期。因此，我们改进的方法可以快速筛选可靠，稳定和高结合亲和力的适体，用于精确的癌症治疗。
Aptamers are short single-stranded DNA or RNA molecules, which have recently been developed for potential broad applications such as clinical therapeutics, diagnosis and tumor-targeted drug delivery. However, the selection of specific aptamers is often unsatisfactory using the classical protein or cell-based SELEX. Herein, we modified the paired cell line approach to identify aptamers targeting leukemia cells expressing the CD33 antigen. Our strategy artfully used the same cells for negative (HEK293T cells) and positive (CD33 transfected-HEK293T cells) aptamer selections, and the negative selections were performed adequately before the positive selection to remove unspecific sequences. The advantages of this strategy are that it is fast and accurate, where only a few rounds of selection together with PCR amplifications are sufficient to obtain high binding affinity antigen-targeted aptamers. By using our modified approach, we successfully obtained the CD33-targeting aptamer S30, which could highly recognize the C2 domain of the CD33 antigen in vitro and in vivo. Moreover, the optimized aptamer S30-T1 (i.e., core region of S30) was conjugated with doxorubicin (Dox) to synthesize S30-T1-Dox conjugates, which could specifically inhibit CD33 positive acute myeloid leukemia HL-60 cell proliferation by arresting the cell cycle at the G2 phase. Thus, our modified approach can rapidly screen reliable, stable and high binding affinity aptamers for precise cancer treatment.