Defective Antiviral Responses of Induced Pluripotent Stem Cells to Baculoviral Vector Transduction
Defective Antiviral Responses of Induced Pluripotent Stem Cells to Baculoviral Vector Transduction
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DOI：
10.1128/jvi.00808-12
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发表时间：
2012-08-01
影响因子：
5.4
通讯作者：
Hu, Yu-Chen
中科院分区：
文献类型：
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作者：
Chen, Guan-Yu;Hwang, Shiaw-Min;Hu, Yu-Chen
诱导多能干细胞（iPSCs）的基因工程对其临床应用具有重要意义，杆状病毒（BV）作为基因传递载体具有广阔的应用前景。为了探索利用BY进行iPSCs转导的可行性，在本研究中，我们首先研究了iPSCs对BV的反应。我们确定BY有效地转导了iPSCs，对细胞增殖、凋亡、多能性和分化没有明显的负面影响。BY的转导轻微扰乱了toll样受体（TLR）信号通路中12个基因的转录，但在蛋白水平上，除了IP-10外，BY没有引发众所周知的细胞因子（如白细胞介素-6 [IL-6]、肿瘤坏死因子α [tnf - α]和β干扰素[ifn - β]）。分子分析显示，iPSCs不表达TLR1、-6、-8或-9，仅表达低水平的TLR2、-3和-4。尽管iPSCs明显表达rig - 1和AIM2等RNA/DNA传感器，但几乎不表达MDA5和DAI （IFN调节因子的DNA依赖性激活因子[IRF]）。重要的是，iPSCs的BY转导没有刺激上述传感器或其下游信号介质（IRF3和NF-kappa B）。这些数据共同证实，由于感知和信号系统受损，iPSCs对BY的反应较差，因此证明了用杆状病毒载体转导iPSCs是合理的。
Genetic engineering of induced pluripotent stem cells (iPSCs) is important for their clinical applications, and baculovirus (BV) holds promise as a gene delivery vector. To explore the feasibility of using BY for iPSCs transduction, in this study we first examined how iPSCs responded to BV. We determined that BY transduced iPSCs efficiently, without inducing appreciable negative effects on cell proliferation, apoptosis, pluripotency, and differentiation. BY transduction slightly perturbed the transcription of 12 genes involved in the Toll-like receptor (TLR) signaling pathway, but at the protein level BY elicited no well-known cytokines (e.g., interleukin-6 [IL-6], tumor necrosis factor alpha [TNF-alpha], and beta interferon [IFN-beta]) except for IP-10. Molecular analyses revealed that iPSCs expressed no TLR1, -6, -8, or -9 and expressed merely low levels of TLR2, -3, and -4. In spite of evident expression of such RNA/DNA sensors as RIG-I and AIM2, iPSCs barely expressed MDA5 and DAI (DNA-dependent activator of IFN regulatory factor [IRF]). Importantly, BY transduction of iPSCs stimulated none of the aforementioned sensors or their downstream signaling mediators (IRF3 and NF-kappa B). These data together confirmed that iPSCs responded poorly to BY due to the impaired sensing and signaling system, thereby justifying the transduction of iPSCs with the baculoviral vector.