在S/MAR序列的活体选择有利于AAV在分裂细胞中的环节维护
Andrea Llanos-Ardaiz1,2, Aquilino Lantero1, Leire Neri1
1Vivet Therapeutics S.L., 31008 Pamplona, Spain.
International journal of molecular sciences
|December 17, 2024
概括
将支架/矩阵附着区域 (S/MAR) 添加到腺相关病毒 (AAV) 载体中,可以改善分裂细胞中的基因组维护. 然而,S/MARs可以降低基因表达,这个问题有可能通过无意中介衰变 (NMD) 抑制序列来解决,以获得更好的基因疗法.
科学领域:
- 基因治疗 基因治疗
- 分子生物学分子生物学
- 肝病学 肝病学是一种肝病学.
背景情况:
- 腺相关病毒 (AAV) 载体对治疗单一性肝病具有前景,但面临的挑战是分裂细胞中的基因组损失,这对于儿科应用至关重要.
- 目前的AAV载体是非整合性的,主要存在于情节,这构成在复制细胞中治疗性基因组稀释的风险.
研究的目的:
- 研究将支架/矩阵附着区域 (S/MAR) 纳入重组AAV (rAAV) 基因组的疗效,以增强分裂细胞中的环节稳定性.
- 评估S/MAR包含对mRNA稳定性和蛋白质表达的影响,并探索减轻潜在负面影响的策略.
主要方法:
- 将含有人类β-干扰素S/MAR序列的rAAV载体给新生儿和婴儿小鼠.
- 病毒基因组水平的量化和mRNA和蛋白质表达的评估.
- 评估与S/MAR元素结合的无意中介衰变 (NMD) 抑制序列.
主要成果:
- 具有S/MAR序列的rAAV载体在新生儿和婴儿小鼠中显示病毒基因组维护的增加.
- 在mRNA的3'端包含S/MARs会对mRNA和蛋白质水平产生负面影响.
- 在含有S/MAR的载体中,无意中介衰变 (NMD) 抑制序列部分恢复了mRNA和蛋白质表达.
结论:
- 脚手架/矩阵附着区域 (S/MARs) 可以改善细胞分裂中的腺相关病毒 (AAV) 载体基因组的持久性,这是儿科基因治疗的一个关键挑战.
- 通过结合NMD抑制序列,可以减轻S/MARs对基因表达的负面影响,为更有效和更持久的基于AAV的基因疗法铺平道路.
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