Conjugation of palmitic acid improves potency and longevity of siRNA delivered via endosomolytic polymer nanoparticles.
Conjugation of palmitic acid improves potency and longevity of siRNA delivered via endosomolytic polymer nanoparticles.
复制标题
DOI：
10.1002/jbm.a.35413
复制
发表时间：
2015-09
期刊：
影响因子：
--
通讯作者：
Duvall CL
中科院分区：
文献类型：
--
作者：
Sarett SM;Kilchrist KV;Miteva M;Duvall CL
siRNA 疗法的临床转化因无法有效克服与细胞内作用生物制剂相关的严格递送障碍而受到限制。在此，为了解决 siRNA 基因沉默的效力和寿命问题，研究了负载有与棕榈酸缀合的 siRNA (siRNA-PA) 的 pH 响应性胶束纳米颗粒 (NP) 载体，作为改善 siRNA 内体逃逸和稳定性的组合方法。相对于未修饰的 siRNA，与疏水性 PA 的缀合提高了 NP 负载效率，从而能够以较低的聚合物：siRNA 比例完全包装 siRNA-PA。 PA 缀合还将核酸货物的细胞内摄取增加了 35 倍，并使细胞内半衰期延长了 3.1 倍。与成纤维细胞和间充质干细胞中的 siRNA NP 相比，对于模型基因荧光素酶和治疗相关基因 PHD2，siRNA-PA NP 的更高摄取和更好的保留与基因沉默 IC50 分别降低 2 倍和 11 倍相关。正如在成纤维细胞中观察到的那样，PA 缀合还显着延长了单次处理后沉默活性的寿命。因此，PA 与 siRNA 的结合与内体溶解 NP 配对是增强 siRNA 在组织再生和其他应用中的功能功效的有前途的方法。
Clinical translation of siRNA therapeutics has been limited by the inability to effectively overcome the rigorous delivery barriers associated with intracellular-acting biologics. Here, in order to address both potency and longevity of siRNA gene silencing, pH-responsive micellar nanoparticle (NP) carriers loaded with siRNA conjugated to palmitic acid (siRNA-PA) were investigated as a combined approach to improve siRNA endosomal escape and stability. Conjugation to hydrophobic PA improved NP loading efficiency relative to unmodified siRNA, enabling complete packaging of siRNA-PA at a lower polymer:siRNA ratio. PA conjugation also increased intracellular uptake of the nucleic acid cargo by 35-fold and produced a 3.1-fold increase in intracellular half-life. The higher uptake and improved retention of siRNA-PA NPs correlated to a 2- and 11-fold decrease in gene silencing IC50 in comparison to siRNA NPs in fibroblasts and mesenchymal stem cells, respectively, for both the model gene luciferase and the therapeutically relevant gene PHD2. PA conjugation also significantly increased longevity of silencing activity following a single treatment, as observed in fibroblasts. Thus, conjugation of PA to siRNA paired with endosomolytic NPs is a promising approach to enhance the functional efficacy of siRNA in tissue regenerative and other applications.