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AAV将RNAi表达构建体递送到非人灵长类动物大脑后的毒性
作者:小柯机器人 发布时间:2021/10/23 20:12:54

美国费城研究所儿童医院Beverly L. Davidson课题组报道了AAV 将RNA 干扰 (RNAi)表达构建体递送到非人灵长类动物大脑后的毒性。相关论文于2021年10月18日发表在《自然—医学》杂志上。

他们证明了递送方法是可扩展的,并且腺相关病毒载体 (AAV.miS1)在短期非人类灵长类动物 (NHP) 试点研究中是安全的。为了将这项技术推向患者,在 NHP 中启动了研究性新药 (IND) 研究。在将 AAV.miS1 递送到小脑深部细胞核后,他们意外地观察到了小脑毒性。小RNA-seq 和使用不含microRNA (miRNA)的 AAV 的研究都表明,这不是内源性 miRNA 加工机制饱和的结果。

RNA-seq 连同 AAV 产品的测序表明,尽管交叉包装的材料数量有限,但仍有大量与神经病理学相关的反向末端重复 (ITR) 启动子活性。通过改变 miS1 表达环境降低了 ITR 启动子活性。他们的啮齿动物和 NHP 研究结果之间令人惊讶的对比突出了在评估用于人类应用的新疗法时对多个物种进行扩展安全性研究的必要性。

据了解,1 型脊髓小脑共济失调的RNAi可以预防和逆转小鼠模型中的行为缺陷和神经病理学读数、安全性和益处持续数月。由AAV.miS1表达的 RNAi 触发器也纠正了错误调节的 miRNA,例如 miR150。

附:英文原文

Title: Toxicity after AAV delivery of RNAi expression constructs into nonhuman primate brain

Author: Keiser, Megan S., Ranum, Paul T., Yrigollen, Carolyn M., Carrell, Ellie M., Smith, Geary R., Muehlmatt, Amy L., Chen, Yong Hong, Stein, Joel M., Wolf, Ronald L., Radaelli, Enrico, Lucas, Timothy J., Gonzalez-Alegre, Pedro, Davidson, Beverly L.

Issue&Volume: 2021-10-18

Abstract: RNA interference (RNAi) for spinocerebellar ataxia type 1 can prevent and reverse behavioral deficits and neuropathological readouts in mouse models, with safety and benefit lasting over many months. The RNAi trigger, expressed from adeno-associated virus vectors (AAV.miS1), also corrected misregulated microRNAs (miRNA) such as miR150. Subsequently, we showed that the delivery method was scalable, and that AAV.miS1 was safe in short-term pilot nonhuman primate (NHP) studies. To advance the technology to patients, investigational new drug (IND)-enabling studies in NHPs were initiated. After AAV.miS1 delivery to deep cerebellar nuclei, we unexpectedly observed cerebellar toxicity. Both small-RNA-seq and studies using AAVs devoid of miRNAs showed that this was not a result of saturation of the endogenous miRNA processing machinery. RNA-seq together with sequencing of the AAV product showed that, despite limited amounts of cross-packaged material, there was substantial inverted terminal repeat (ITR) promoter activity that correlated with neuropathologies. ITR promoter activity was reduced by altering the miS1 expression context. The surprising contrast between our rodent and NHP findings highlight the need for extended safety studies in multiple species when assessing new therapeutics for human application. A preclinical safety study of adeno-associated viruses (AAVs) for RNA interference (RNAi) for spinocerebellar ataxia type 1 therapy showed toxicity in nonhuman primates but not rodents, due to unexpected AAV inverted terminal repeat transcriptional activity that was mitigated on altering the RNAi expression environment.

DOI: 10.1038/s41591-021-01522-3

Source: https://www.nature.com/articles/s41591-021-01522-3

期刊信息

Nature Medicine:《自然—医学》,创刊于1995年。隶属于施普林格·自然出版集团,最新IF:30.641
官方网址:https://www.nature.com/nm/
投稿链接:https://mts-nmed.nature.com/cgi-bin/main.plex