Selection of a Novel Aptamer Against Vitronectin Using Capillary Electrophoresis and Next Generation Sequencing.
Selection of a Novel Aptamer Against Vitronectin Using Capillary Electrophoresis and Next Generation Sequencing.
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使用毛细管电泳和下一代测序选择一种针对玻璃体素的新型适体。
DOI：
10.1038/mtna.2016.91
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发表时间：
2016-11-15
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影响因子：
--
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乳腺癌（BC）在美国每年导致约40，000人死亡，即使在幸存者中，对该疾病的治疗也可能产生破坏性后果，包括心脏病风险增加和化疗毒性作用导致的认知障碍。适体介导的药物递送可以通过选择性地将化疗递送至BC细胞而有助于改善治疗结果，前提是可以鉴定合适的癌症特异性抗原。我们在这里报告使用毛细管电泳结合下一代测序开发第一玻连蛋白（VN）结合适体（Kd 405 nmol/l，第一个与玻连蛋白（VN; Kd = 405 nmol/l），一种在伤口愈合中起重要作用的蛋白质，并且相对于在BC组织中的蛋白质，其在BC组织和BC患者的血液中以升高的水平存在。相应的非恶性组织。我们使用Dox-01开发了DVBA-01，一种二聚体适体复合物，并使用pH敏感的共价键将多柔比星（Dox）与DVBA-01缀合（7：1比例）。Dox缀合增强了复合物的热稳定性（60.2 ° C对46.5°C），并且不降低对VN靶标的亲和力。所得DVBA-01-Dox复合物对在VN（1.8 × 10− 6 mol/l）包被的塑料器皿上培养的MDA-MB-231 BC细胞的细胞毒性相对于未包被的平板（2.4 × 10− 6 mol/l）或相关蛋白纤连蛋白包被的平板（2.1 × 10−6 mol/l）增加。使用免疫组织化学评价了EST-O 1适体与人BC组织的结合，并显示出与BC细胞的组织特异性结合和明显缔合。相反，优先结合多聚体VN的单克隆抗体主要染色BC组织的细胞外基质和血管壁。我们的研究结果表明，使用VN靶向适体来改善药物递送以治疗BC的潜力很大。
Breast cancer (BC) results in ~40,000 deaths each year in the United States and even among survivors treatment of the disease may have devastating consequences, including increased risk for heart disease and cognitive impairment resulting from the toxic effects of chemotherapy. Aptamer-mediated drug delivery can contribute to improved treatment outcomes through the selective delivery of chemotherapy to BC cells, provided suitable cancer-specific antigens can be identified. We report here the use of capillary electrophoresis in conjunction with next generation sequencing to develop the first vitronectin (VN) binding aptamer (VBA-01; Kd 405 nmol/l, the first aptamer to vitronectin (VN; Kd = 405 nmol/l) , a protein that plays an important role in wound healing and that is present at elevated levels in BC tissue and in the blood of BC patients relative to the corresponding nonmalignant tissues. We used VBA-01 to develop DVBA-01, a dimeric aptamer complex, and conjugated doxorubicin (Dox) to DVBA-01 (7:1 ratio) using pH-sensitive, covalent linkages. Dox conjugation enhanced the thermal stability of the complex (60.2 versus 46.5°C) and did not decrease affinity for the VN target. The resulting DVBA-01-Dox complex displayed increased cytotoxicity to MDA-MB-231 BC cells that were cultured on plasticware coated with VN (1.8 × 10−6mol/l) relative to uncoated plates (2.4 × 10−6 mol/l), or plates coated with the related protein fibronectin (2.1 × 10−6 mol/l). The VBA-01 aptamer was evaluated for binding to human BC tissue using immunohistochemistry and displayed tissue specific binding and apparent association with BC cells. In contrast, a monoclonal antibody that preferentially binds to multimeric VN primarily stained extracellular matrix and vessel walls of BC tissue. Our results indicate a strong potential for using VN-targeting aptamers to improve drug delivery to treat BC.
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影响因子：
45.3
作者：
Cristofanilli, M;Hayes, DF;Terstappen, LWMM
通讯作者：
Terstappen, LWMM
影响因子：
4.3
作者：
Riley, Kathryn R.;Gagliano, Jason;Xiao, Jiajie;Libby, Kara;Saito, Shingo;Yu, Guo;Cubicciotti, Roger;Macosko, Jed;Colyer, Christa L.;Guthold, Martin;Bonin, Keith
通讯作者：
Bonin, Keith
影响因子：
4.1
作者：
Riley, Kathryn R.;Saito, Shingo;Colyer, Christa L.
通讯作者：
Colyer, Christa L.
DOI：
10.1083/jcb.200612058
发表时间：
2007-06-04
期刊：
The Journal of cell biology
影响因子：
--
作者：
Madsen CD;Ferraris GM;Andolfo A;Cunningham O;Sidenius N
通讯作者：
Sidenius N
影响因子：
6
作者：
Jo, Minji;Takimoto, Shinako;Gonias, Steven L.
通讯作者：
Gonias, Steven L.