One-year follow-up of gene expression by integrase-defective lentiviral vectors and their therapeutic potential in spinocerebellar ataxia model mice.
One-year follow-up of gene expression by integrase-defective lentiviral vectors and their therapeutic potential in spinocerebellar ataxia model mice.
复制标题
整合酶缺陷型慢病毒载体的基因表达及其在脊髓小脑共济失调模型小鼠中的治疗潜力的一年随访。
DOI：
10.1038/gt.2014.60
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发表时间：
2014
期刊：
影响因子：
5.1
通讯作者：
and Hirai H.
中科院分区：
文献类型：
--
作者：
Saida H;Matsuzaki Y;Takayama K;Iizuka A;Konno A;Yanagi S;and Hirai H.
我们研究了整合酶缺陷慢病毒载体（idlv）与整合酶突变体（D64V）的剩余整合能力、转基因表达水平和持续时间，以及与整合酶基因野生型慢病毒载体（WTLVs）的治疗潜力。与WTLVs相比，idlv介导的前病毒整合到宿主细胞染色体的比例在HeLa细胞中约为1/3850，在小鼠小脑神经元中约为1/111。在2个月时，idlv在小鼠小脑中的转基因表达与WTLVs相当，但随后显著降低。注射后6个月和12个月，idlv感染小脑的mRNA水平分别约为wtlv注射小脑的26%和5%。为了研究其治疗潜力，将表达一种增强泛素-蛋白酶体途径的分子的idlv或WTLVs注射到脊髓小脑性共济失调3型模型小鼠（SCA3小鼠）的小脑中。注射idlv的SCA3小鼠即使在注射后1年也表现出明显改善的旋转杆性能。注射后1年的免疫组织化学显示，注射idlv和注射wtlv的SCA3小鼠浦肯野细胞中突变体聚集量急剧减少。我们的研究结果表明，由于插入突变的风险大大降低，idlv作为基因治疗载体是更安全、潜在有效的。
We examined integrase-defective lentiviral vectors (IDLVs) with a mutant (D64V) integrase in terms of their residual integration capability, the levels and duration of transgene expression and their therapeutic potential in comparison to wild-type lentiviral vectors (WTLVs) with a wild-type integrase gene. Compared with WTLVs, the IDLV-mediated proviral integration into host-cell chromosomes was approximately 1/3850 in HeLa cells and approximately 1/111 in mouse cerebellar neurons in vivo. At 2 months, transgene expression by IDLVs in the mouse cerebellum was comparable to that by WTLVs, but then significantly decreased. The mRNA levels at 6 and 12 months after injection in IDLV-infected cerebella were approximately 26% and 5%, respectively, of the mRNA levels in WTLV-injected cerebella. To examine the therapeutic potential, IDLVs or WTLVs expressing a molecule that enhances the ubiquitin-proteasome pathway were injected into the cerebella of spinocerebellar ataxia type 3 model mice (SCA3 mice). IDLV-injected SCA3 mice showed a significantly improved rotarod performance even at 1 year after-injection. Immunohistochemistry at 1 year after injection showed a drastic reduction of mutant aggregates in Purkinje cellsfrom IDLV-injected, as well as WTLV-injected, SCA3 mice. Our results suggest that because of the substantially reduced risk of insertional mutagenesis, IDLVs are safer and potentially effective as gene therapy vectors.