GFAP Promoter-Driven RNA Interference on TGF-β1 to Treat Liver Fibrosis
GFAP Promoter-Driven RNA Interference on TGF-β1 to Treat Liver Fibrosis
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DOI：
10.1007/s11095-011-0384-y
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发表时间：
2011-04-01
影响因子：
3.7
通讯作者：
Mahato, Ram I.
中科院分区：
文献类型：
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作者：
Yang, Ningning;Mahato, Ram I.
本研究的目的是确定启动子和miRNA骨架在基于shRNA的肝星状细胞(HSC)特异性转化生长因子-β1基因沉默中的作用。这有望避免非特异性的转化生长因子-β1基因沉默的副作用。将针对大鼠转化生长因子-β1基因769和1033个起始点的两个最有效的shRNA克隆到pSilencer 1.0载体中，以增强转化生长因子-β1基因的沉默。然后，我们构建了HSC特异性的pri-miRNA模拟和Pri-miRNA簇模拟表达载体，其中shRNA的表达由胶质纤维酸性蛋白(GFAP)启动子驱动，从而实现HSC特异性的转化生长因子-β1基因沉默，从而避免对其他细胞和组织中转化生长因子-β1表达的非特异性抑制。转化生长因子-β1 Pri-miRNA簇模拟载体可在mRNA和蛋白水平下调转化生长因子-β1和胶原基因的表达，GFAP启动子驱动的转化生长因子-β1 Pri-miRNA产生载体有望用于肝纤维化的定点基因治疗。
The objective was to determine the role of promoters and miRNA backbone in shRNA-based hepatic stellate cell (HSC)-specific transforming growth factor (TGF)-beta 1 gene silencing. This is expected to avoid the side effect of non-specific TGF-beta 1 gene silencing.Two most potent shRNAs targeting 769 and 1033 start sites of rat TGF-beta 1 mRNA were cloned into pSilencer 1.0 vector for enhanced TGF-beta 1 gene silencing. We then constructed HSC-specific pri-miRNA mimic and pri-miRNA cluster mimic expression plasmids in which shRNA expression was driven by a glial fibrillary acidic protein (GFAP) promoter to achieve HSC-specific TGF-beta 1 gene silencing to avoid nonspecific inhibition of TGF-beta 1 expression in other cells and organs.These TGF-beta 1 pri-miRNA-producing plasmids showed the inhibition of proliferation and induced apoptosis of activated HSC-T6 cells. TGF-beta 1 pri-miRNA cluster mimic plasmids decreased TGF-beta 1 and collagen gene expression at both mRNA and protein levels.GFAP promoter driven TGF-beta 1 pri-miRNA producing plasmids have the potential to be used for site-specific gene therapeutics to treat liver fibrosis.