A New Transferrin Receptor Aptamer Inhibits New World Hemorrhagic Fever Mammarenavirus Entry
A New Transferrin Receptor Aptamer Inhibits New World Hemorrhagic Fever Mammarenavirus Entry
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DOI：
10.1038/mtna.2016.32
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发表时间：
2016-05-24
影响因子：
8.8
通讯作者：
Levy, Matthew
中科院分区：
文献类型：
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作者：
Maier, Keith E.;Jangra, Rohit K.;Levy, Matthew
致病性新世界出血热乳房病毒 (NWM) 利用糖蛋白 1 (GP1) 靶向人转铁蛋白受体 (hTfR) 的顶端结构域，以促进细胞进入。然而，它们的 GP1 之间的保守性很低。考虑到这一点以及哺乳动物与病毒相比缓慢的进化进程，hTfR 的治疗靶向为交叉菌株抑制和减少逃逸突变体的可能性提供了一条有吸引力的途径。适体对于开发进入小瓶的抑制剂具有独特的优势，包括易于合成、缺乏免疫原性以及潜在的打破南美流行疾病冷链的解决方案。在这里，我们认识到与天然配体转铁蛋白 (Tf) 的体内竞争可能推动 GP1 进化以识别顶端结构域，因此我们对补充了 Tf 的 hTfR 表达细胞进行了体外竞争选择。所得的最小化适体 Waz 与受体的顶端结构域结合，并抑制培养物中重组 NWM 对人类细胞的感染（EC50 类似于 400 nmol/l）。适体多聚化进一步增强了抑制作用 >10 倍（EC50 类似于 30 nmol/l）。总之，我们的结果强调了使用竞争对手来偏向选择结果的能力，并证明了如何利用亲合力效应来增强适体结合和病毒抑制效力。
Pathogenic New World hemorrhagic fever mammarenaviruses (NWM) utilize Glycoprotein 1 (GP1) to target the apical domain of the human transferrin receptor (hTfR) for facilitating cell entry. However, the conservation between their GP1s is low. Considering this and the slow evolutionary progression of mammals compared to viruses, therapeutic targeting of hTfR provides an attractive avenue for cross-strain inhibition and diminishing the likelihood of escape mutants. Aptamers present unique advantages for the development of inhibitors to vial entry, including ease of synthesis, lack of immunogenicity, and potentially cold-chain breaking solutions to diseases endemic to South America. Here, recognizing that in vivo competition with the natural ligand, transferrin (Tf), likely drove the evolution of GP1 to recognize the apical domain, we performed competitive in vitro selections against hTfR-expressing cells with supplemented Tf. The resultant minimized aptamer, Waz, binds the apical domain of the receptor and inhibits infection of human cells by recombinant NWM in culture (EC50 similar to 400 nmol/l). Aptamer multimerization further enhanced inhibition >10-fold (EC50 similar to 30 nmol/l). Together, our results highlight the ability to use a competitor to bias the outcome of a selection and demonstrate how avidity effects can be leveraged to enhance both aptamer binding and the potency of viral inhibition.