LNA-mediated microRNA silencing in non-human primates
LNA-mediated microRNA silencing in non-human primates
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DOI：
10.1038/nature06783
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发表时间：
2008-04-17
期刊：
影响因子：
64.8
通讯作者：
Kauppinen, Sakari
中科院分区：
文献类型：
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作者：
Elmen, Joacim;Lindow, Morten;Kauppinen, Sakari
microRNAs （miRNAs）是在发育和疾病中起重要作用的小调控rna(1-3)，因此代表了治疗干预的潜在新靶点(4)。尽管最近在啮齿类动物中mirna的沉默方面取得了进展（5,6），但在体内开发有效且安全的序列特异性mirna拮抗方法仍然是一项重大的科学和治疗挑战。此外，在灵长类动物中没有miRNA拮抗剂的报道。在这里，我们展示了简单的系统递送非偶联的，PBS配制的锁定核酸修饰的寡核苷酸（LNA- anti - ir）有效地拮抗非人类灵长类动物肝脏表达的miR- 122。急性静脉注射3或10 mg kg(-1) LNA-anti - ir给非洲绿猴，导致LNA-anti - ir在灵长类动物肝细胞的细胞质中被摄取，并在LNA-anti - ir和miR- 122之间形成稳定的异双链。这伴随着成熟miR- 122的消耗和血浆胆固醇的剂量依赖性降低。在灵长类动物中，通过三次剂量的10mg kg(-1) LNA- anti - ir实现了miR- 122的有效沉默，导致血浆总胆固醇的持久和可逆下降，而没有证据表明LNA相关的毒性或组织病理学变化。我们的研究结果证明了系统给药LNA- antimiRs在探索啮齿动物和灵长类动物miRNA功能方面的实用性，并支持这些化合物作为疾病相关miRNA的新一类治疗药物的潜力。
microRNAs ( miRNAs) are small regulatory RNAs that are important in development and disease(1-3) and therefore represent a potential new class of targets for therapeutic intervention(4). Despite recent progress in silencing of miRNAs in rodents(5,6), the development of effective and safe approaches for sequence-specific antagonism of miRNAs in vivo remains a significant scientific and therapeutic challenge. Moreover, there are no reports of miRNA antagonism in primates. Here we show that the simple systemic delivery of a unconjugated, PBS- formulated locked- nucleic- acid- modified oligonucleotide ( LNA- antimiR) effectively antagonizes the liver- expressed miR- 122 in non- human primates. Acute administration by intravenous injections of 3 or 10 mg kg(-1) LNA- antimiR to African green monkeys resulted in uptake of the LNA- antimiR in the cytoplasm of primate hepatocytes and formation of stable heteroduplexes between the LNA-antimiR and miR- 122. This was accompanied by depletion of mature miR- 122 and dose- dependent lowering of plasma cholesterol. Efficient silencing of miR- 122 was achieved in primates by three doses of 10 mg kg(-1) LNA- antimiR, leading to a long- lasting and reversible decrease in total plasma cholesterol without any evidence for LNA- associated toxicities or histopathological changes in the study animals. Our findings demonstrate the utility of systemically administered LNA- antimiRs in exploring miRNA function in rodents and primates, and support the potential of these compounds as a new class of therapeutics for disease-associated miRNAs.