Ghent Research Group on Nanomedicines, Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmaceutical Sciences, Department of Pharmaceutics, Ghent University, Ottergemsesteenweg 460, 9000 Ghent, Belgium.
VIB-UGent, Center for Medical Biotechnology, Technologiepark 927, 9052 Zwijnaarde (Ghent), Belgium; Department of Biomedical Molecular Biology, Ghent University, Technologiepark 927, 9052 Zwijnaarde (Ghent), Belgium.
Acta Biomater. 2018 Sep 15;78:236-246. doi: 10.1016/j.actbio.2018.08.012. Epub 2018 Aug 15.
Despite the many advantages of small interfering RNA (siRNA) inhalation therapy and a growing prevalence of respiratory pathologies, its clinical translation is severely hampered by inefficient intracellular delivery. To this end, we previously developed hybrid nanoparticles consisting of an siRNA-loaded nanosized hydrogel core (nanogel) coated with Curosurf®, a clinically used pulmonary surfactant (PS). Interestingly, the PS shell was shown to markedly improve particle stability as well as intracellular siRNA delivery in vitro and in vivo. The major aim of this work was to identify the key molecular components of PS responsible for the enhanced siRNA delivery and evaluate how the complexity of the PS coat could be reduced. We identified surfactant protein B (SP-B) as a potent siRNA delivery enhancer when reconstituted in proteolipid coated hydrogel nanocomposites. Improved cytosolic siRNA delivery was achieved by inserting SP-B into a simplified phospholipid mixture prior to nanogel coating. This effect was observed both in vitro (lung epithelial cell line) and in vivo (murine acute lung injury model), albeit that distinct phospholipids were required to achieve these results. Importantly, the developed nanocomposites have a low in vivo toxicity and are efficiently taken up by resident alveolar macrophages, a main target cell type for treatment of inflammatory pulmonary pathologies. Our results demonstrate the potential of the endogenous protein SP-B as an intracellular siRNA delivery enhancer, paving the way for future design of nanoformulations for siRNA inhalation therapy.
Despite the therapeutic potential of small interfering RNA (siRNA) and a growing prevalence of lung diseases for which innovative therapies are needed, a safe and effective siRNA inhalation therapy remains non-existing due to a lack of suitable formulations. We identified surfactant protein B (SP-B) as a potent enhancer of siRNA delivery by proteolipid coated nanogel formulations in vitro in a lung epithelial cell line. The developed nanocomposites have a low in vivo toxicity and show a high uptake by alveolar macrophages, a main target cell type for treatment of inflammatory pulmonary pathologies. Importantly, in vivo SP-B is also critical for the developed formulation to obtain a significant silencing of TNFα in a murine LPS-induced acute lung injury model.
尽管小干扰 RNA(siRNA)吸入疗法有许多优势，且呼吸道疾病的发病率也在不断上升，但由于细胞内递药效率低下，其临床转化仍受到严重阻碍。为此，我们之前开发了由负载 siRNA 的纳米水凝胶核（纳米凝胶）和一种临床应用的肺表面活性剂（PS）Curosurf®组成的混合纳米粒。有趣的是，PS 壳明显提高了颗粒稳定性以及体外和体内的细胞内 siRNA 递药效率。本工作的主要目的是确定 PS 中负责增强 siRNA 递药的关键分子成分，并评估 PS 涂层的复杂性如何降低。我们发现，当重新构成包被有蛋白的水凝胶纳米复合物时，表面活性蛋白 B（SP-B）是一种有效的 siRNA 递药增强剂。通过在纳米凝胶包被前将 SP-B 插入简化的磷脂混合物中，可实现细胞质中 siRNA 的有效递药。这一效应不仅在体外（肺上皮细胞系），而且在体内（小鼠急性肺损伤模型）都观察到了，尽管实现这些结果需要不同的磷脂。重要的是，所开发的纳米复合物具有低体内毒性，并被驻留的肺泡巨噬细胞有效摄取，这是治疗炎症性肺疾病的主要靶细胞类型。我们的结果表明，内源性蛋白 SP-B 具有作为细胞内 siRNA 递药增强剂的潜力，为未来设计用于 siRNA 吸入疗法的纳米制剂铺平了道路。
尽管小干扰 RNA（siRNA）具有治疗潜力，且需要创新疗法的肺部疾病患病率不断上升，但由于缺乏合适的制剂，安全有效的 siRNA 吸入疗法仍不存在。我们在体外肺上皮细胞系中发现，表面活性蛋白 B（SP-B）通过包被有蛋白的纳米凝胶制剂作为 siRNA 递药的有效增强剂。所开发的纳米复合物具有低体内毒性，并被肺泡巨噬细胞高效摄取，肺泡巨噬细胞是治疗炎症性肺疾病的主要靶细胞类型。重要的是，体内 SP-B 对于开发的制剂在 LPS 诱导的小鼠急性肺损伤模型中获得 TNFα 的显著沉默也至关重要。
