Development of Non-Viral, Trophoblast-Specific Gene Delivery for Placental Therapy.
Development of Non-Viral, Trophoblast-Specific Gene Delivery for Placental Therapy.
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DOI：
10.1371/journal.pone.0140879
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Jones H
中科院分区：
文献类型：
--
作者：
Abd Ellah N;Taylor L;Troja W;Owens K;Ayres N;Pauletti G;Jones H
低出生体重既与短期问题有关，也与胎儿对成人发病疾病的规划有关，包括肥胖、糖尿病和心血管疾病的风险增加。胎盘功能不全可导致胎儿宫内生长受限(IUGR)，导致发育性疾病的流行。目前还没有治疗IUGR或胎盘功能不全的方法。为了解决这一问题并朝着宫内治疗的方向发展，我们采用了一种与IGF-1基因复合的纳米结构递送系统来治疗胎盘。IGF-1是一种生长因子，对实现适当的胎盘和胎儿生长至关重要。在滋养层特异性启动子(Cyp19a或PLAC1)的控制下，利用与hIGF-1质粒DNA络合的两嵌段共聚物(PHPMA-b-pDMAEMA)将基因输送到人滋养层细胞和小鼠胎盘模型中。通过荧光显微镜观察pEGFP-C1纳米载体在BeWo细胞和非滋养细胞中的转染率。通过胎盘直接注射到IUGR小鼠模型中来评估体内的转染性和功能性。用PHPMA-b-pDMAEMA与CyP19a-923或PLAC1修饰的质粒形成的复合体在体外诱导滋养层细胞选择性转基因表达，胎盘注射PLAC1-hIGF-1在我们的小鼠模型中产生可测量的RNA表达并缓解IUGR，从而代表了人类胎盘基因治疗的创新构建块。
Low birth weight is associated with both short term problems and the fetal programming of adult onset diseases, including an increased risk of obesity, diabetes and cardiovascular disease. Placental insufficiency leading to intrauterine growth restriction (IUGR) contributes to the prevalence of diseases with developmental origins. Currently there are no therapies for IUGR or placental insufficiency. To address this and move towards development of an in utero therapy, we employ a nanostructure delivery system complexed with the IGF-1 gene to treat the placenta. IGF-1 is a growth factor critical to achieving appropriate placental and fetal growth. Delivery of genes to a model of human trophoblast and mouse placenta was achieved using a diblock copolymer (pHPMA-b-pDMAEMA) complexed to hIGF-1 plasmid DNA under the control of trophoblast-specific promoters (Cyp19a or PLAC1). Transfection efficiency of pEGFP-C1-containing nanocarriers in BeWo cells and non-trophoblast cells was visually assessed via fluorescence microscopy. In vivo transfection and functionality was assessed by direct placental-injection into a mouse model of IUGR. Complexes formed using pHPMA-b-pDMAEMA and CYP19a-923 or PLAC1-modified plasmids induce trophoblast-selective transgene expression in vitro, and placental injection of PLAC1-hIGF-1 produces measurable RNA expression and alleviates IUGR in our mouse model, consequently representing innovative building blocks towards human placental gene therapies.