Implementing Efficient Peptoid-Mediated Delivery of RNA-Based Therapeutics to the Vocal Folds.
Implementing Efficient Peptoid-Mediated Delivery of RNA-Based Therapeutics to the Vocal Folds.
复制标题
实施基于 RNA 的治疗剂向声带的有效肽介导递送。
DOI：
10.1002/lio2.310
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发表时间：
2019
影响因子：
1.9
通讯作者：
Branski,RyanC
中科院分区：
文献类型：
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作者：
Mukudai,Shigeyuki;Kraja,Iv;Bing,Renjie;Nalband,DanielleM;Tatikola,Mallika;Hiwatashi,Nao;Kirshenbaum,Kent;Branski,RyanC
我们假设Smad 3是声带（VF）纤维化的主要调节因子，靶向Smad 3的基于RNA的治疗方法具有治疗前景。交付仍然具有挑战性。我们以前描述了一种新的合成类肽寡聚体lipitoidL 0，与siRNA复合，以提高稳定性和细胞摄取。然而，这些类肽的优点是极大的结构和化学延展性，以优化转染效率。L0的修饰进行了测定，以优化siRNA介导的基因expression.MethodsIn体外，Smad 3敲低各种类脂质变体进行了评估，通过定量真实的时间聚合酶链反应在人VF成纤维细胞。通过比色测定法定量细胞毒性。在体内，VF损伤的兔模型被用来评估注射theL 0-siRNA complex.ResultsIn vitro后Smad 3敲低的时间动力学，建立了类似的减少Smad 3表达的所有类脂质变体，有一个例外。序列变体也表现出相似的无毒特征;未观察到细胞增殖的统计学显著差异。在体内，注射L0复合的Smad 3 siRNA后1天，损伤的VF中Smad 3表达显著降低。Smad 3表达的定性抑制持续到3天后的损伤，但没有达到统计学significant.ConclusionsIn尽管这些类肽转染试剂的化学多样性，序列变异体一般提供一致的有效减少Smad 3 expressions.L0产生了有效的，但暂时有限的敲低Smad 3在体内。类肽可以为发现针对临床应用优化的siRNA递送载体提供多功能平台。
ObjectiveWe hypothesize that Smad3 is a master regulator of fibrosis in the vocal folds (VFs) and RNA‐based therapeutics targeting Smad3 hold therapeutic promise. Delivery remains challenging. We previously described a novel synthetic peptoid oligomer, lipitoidL0, complexed with siRNA to improve stability and cellular uptake. An advantage of these peptoids, however, is tremendous structural and chemical malleability to optimize transfection efficiency. Modifications ofL0were assayed to optimize siRNA‐mediated alteration of gene expression.MethodsIn vitro, Smad3 knockdown by various lipitoid variants was evaluated via quantitative real‐time polymerase chain reaction in human VF fibroblasts. Cytotoxicity was quantified via colorimetric assays. In vivo, a rabbit model of VF injury was employed to evaluate the temporal dynamics of Smad3 knockdown following injection of theL0‐siRNA complex.ResultsIn vitro, similar reductions in Smad3 expression were established by all lipitoid variants, with one exception. Sequence variants also exhibited similar nontoxic characteristics; no statistically significant differences in cell proliferation were observed. In vivo, Smad3 expression was significantly reduced in injured VFs following injection ofL0‐complexed Smad3 siRNA at 1 day postinjection. Qualitative suppression of Smad3 expression persisted to 3 days following injury, but did not achieve statistical significance.ConclusionsIn spite of the chemical diversity of these peptoid transfection reagents, the sequence variants generally provided consistently efficient reductions in Smad3 expression.L0yielded effective, yet temporally limited knockdown of Smad3 in vivo. Peptoids may provide a versatile platform for the discovery of siRNA delivery vehicles optimized for clinical application.Level of EvidenceNA