Kraja Iv, Bing Renjie, Hiwatashi Nao, Rousseau Bernard, Nalband Danielle, Kirshenbaum Kent, Branski Ryan C
NYU Voice Center, Department of Otolaryngology-Head and Neck Surgery, New York University School of Medicine, New York, New York, U.S.A.
Bill Wilkerson Center for Otolaryngology and Communication Sciences, Department of Otolaryngology, Hearing and Speech Sciences, and Mechanical Engineering, Vanderbilt University Medical Center, Nashville, Tennessee, U.S.A.
Laryngoscope. 2017 Jul;127(7):E231-E237. doi: 10.1002/lary.26432. Epub 2016 Dec 20.
An obstacle to clinical use of RNA-based gene suppression is instability and inefficiency of current delivery modalities. Nanoparticle delivery likely holds great promise, but the kinetics and transfection conditions must be optimized prior to in vivo utility. We investigated a RNA nanoparticle complex incorporating a lipitoid transfection reagent in comparison to a commercially available reagent.
In vitro.
We investigated which variables influence transfection efficiency of lipitoid oligomers and a commercially available reagent across species, in vitro. These variables included duration, dose, and number of administrations, as well as serum and media conditions. The target gene was Smad3, a signaling protein in the transforming growth factor-β cascade implicated in fibroplasia in the vocal folds and other tissues.
The two reagents suppressed Smad3 mRNA for up to 96 hours; lipitoid performed favorably and comparably. Both compounds yielded 60% to 80% mRNA knockdown in rat, rabbit, and human vocal fold fibroblasts (P < 0.05 relative to control). Dose and number of administrations played a significant role in gene suppression (P < 0.05). Suppression was more dose-sensitive with lipitoid. At a constant siRNA concentration, a 50% decrease in gene expression was observed in response to a five-fold increase in lipitoid concentration. Increased number of administrations enhanced gene suppression, ∼45% decrease between one and four administrations. Neither serum nor media type altered efficiency.
Lipitoid effectively knocked down Smad3 expression across multiple transfection conditions. These preliminary data are encouraging, and lipitoid warrants further investigation with the goal of clinical utility.
NA. Laryngoscope, 127:E231-E237, 2017.
基于RNA的基因抑制技术在临床应用中的一个障碍是当前递送方式的不稳定性和低效性。纳米颗粒递送可能具有很大的前景，但在体内应用之前，必须优化其动力学和转染条件。我们研究了一种包含类脂质转染试剂的RNA纳米颗粒复合物，并与一种市售试剂进行了比较。
体外研究。
我们在体外研究了哪些变量会影响类脂质寡聚物和市售试剂在不同物种中的转染效率。这些变量包括作用持续时间、剂量、给药次数，以及血清和培养基条件。目标基因是Smad3，它是转化生长因子-β信号级联反应中的一种信号蛋白，与声带及其他组织的纤维增生有关。
两种试剂均可抑制Smad3 mRNA达96小时；类脂质的表现良好且与之相当。两种化合物在大鼠、兔和人声带成纤维细胞中均使mRNA敲低60%至80%（相对于对照组，P < 0.05）。剂量和给药次数在基因抑制中起显著作用（P < 0.05）。类脂质对剂量更为敏感。在siRNA浓度恒定的情况下，随着类脂质浓度增加五倍，基因表达下降50%。给药次数增加可增强基因抑制效果，一次给药和四次给药之间基因表达下降约45%。血清和培养基类型均未改变转染效率。
类脂质在多种转染条件下均能有效敲低Smad3表达。这些初步数据令人鼓舞，类脂质值得进一步研究以期用于临床。
无。《喉镜》，2017年，第127卷，E231 - E237页
