Chemically Modified, α-Amino-3-hydroxy-5-methyl-4-isoxazole (AMPA) Receptor RNA Aptamers Designed for in Vivo Use.
Chemically Modified, α-Amino-3-hydroxy-5-methyl-4-isoxazole (AMPA) Receptor RNA Aptamers Designed for in Vivo Use.
复制标题
经过化学修饰的 α-氨基-3-羟基-5-甲基-4-异恶唑 (AMPA) 受体 RNA 适体设计用于体内使用。
DOI：
10.1021/acschemneuro.7b00211
复制
发表时间：
2017
影响因子：
5
通讯作者：
Niu,Li
中科院分区：
文献类型：
--
作者：
Huang,Zhen;Wen,Wei;Wu,Andrew;Niu,Li
谷氨酸离子通道有三种亚型，即α-氨基-3-羟基-5-甲基-4-异恶唑（AMPA）、海因酸盐和n -甲基-d-天冬氨酸（NMDA）受体。这些受体亚型单独或集体的过度活动与各种神经系统疾病有关。RNA适体作为这些受体的拮抗剂是潜在的治疗方法。为了开发适体疗法，必须对RNA适体进行化学修饰，使其在生物液体中具有核糖核酸酶抗性或稳定性。利用配体的系统进化指数富集（SELEX）和化学修饰文库，酶法制备（即，文库包含2 ' -氟修饰的核苷三磷酸或atp， ctp和UTPs，但常规gtp），我们已经分离出一个适体。短适配体（69个核苷酸）FN1040选择性抑制GluA1和GluA2QflipAMPA受体亚基，而全长适配体（101个核苷酸）FN1040还抑制GluK1，但不抑制kainate受体GluK2，以及GluN1a/2A和GluN1a/2B这两种主要的天然NMDA受体。两种适体在含血清的培养基或脑脊液中表现出相似的效价（2 ~ 4 μM），半衰期至少为2天。因此，这两个适体对病毒是可适应的。
Glutamate ion channels have three subtypes, that is, α-amino-3-hydroxy-5-methyl-4-isoxazole (AMPA), kainate, andN-methyl-d-aspartate (NMDA) receptors. Excessive activity of these receptor subtypes either individually or collectively is involved in various neurological disorders. RNA aptamers as antagonists of these receptors are potential therapeutics. For developing aptamer therapeutics, the RNA aptamers must be chemically modified to become ribonuclease-resistant or stable in biological fluids. Using systematic evolution of ligands by exponential enrichment (SELEX) and a chemically modified library, prepared enzymatically (i.e., the library contains RNAs with 2′-fluoro modified nucleoside triphosphates or ATPs, CTPs and UTPs, but regular GTPs), we have isolated an aptamer. The short aptamer (69 nucleotides) FN1040s selectively inhibits the GluA1 and GluA2QflipAMPA receptor subunits, whereas the full-length aptamer (101 nucleotides) FN1040 additionally inhibits GluK1, but not GluK2, kainate receptor, and GluN1a/2A and GluN1a/2B, the two major native NMDA receptors. The two aptamers show similar potency (2–4 μM) and are stable with a half-life of at least 2 days in serum-containing medium or cerebrospinal fluid. Therefore, these two aptamers are amenable forin vivouse.