Department of Neurosurgery, Sheba Medical Center , Ramat Gan, Israel.
ACS Nano. 2015 Feb 24;9(2):1581-91. doi: 10.1021/nn506248s. Epub 2015 Jan 8.
Glioblastoma multiforme (GBM) is one of the most infiltrating, aggressive, and poorly treated brain tumors. Progress in genomics and proteomics has paved the way for identifying potential therapeutic targets for treating GBM, yet the vast majority of these leading drug candidates for the treatment of GBM are ineffective, mainly due to restricted passages across the blood-brain barrier. Nanoparticles have been emerged as a promising platform to treat different types of tumors due to their ability to transport drugs to target sites while minimizing adverse effects. Herein, we devised a localized strategy to deliver RNA interference (RNAi) directly to the GBM site using hyaluronan (HA)-grafted lipid-based nanoparticles (LNPs). These LNPs having an ionized lipid were previously shown to be highly effective in delivering small interfering RNAs (siRNAs) into various cell types. LNP's surface was functionalized with hyaluronan (HA), a naturally occurring glycosaminoglycan that specifically binds the CD44 receptor expressed on GBM cells. We found that HA-LNPs can successfully bind to GBM cell lines and primary neurosphers of GBM patients. HA-LNPs loaded with Polo-Like Kinase 1 (PLK1) siRNAs (siPLK1) dramatically reduced the expression of PLK1 mRNA and cumulated in cell death even under shear flow that simulate the flow of the cerebrospinal fluid compared with control groups. Next, a human GBM U87MG orthotopic xenograft model was established by intracranial injection of U87MG cells into nude mice. Convection of Cy3-siRNA entrapped in HA-LNPs was performed, and specific Cy3 uptake was observed in U87MG cells. Moreover, convection of siPLK1 entrapped in HA-LNPs reduced mRNA levels by more than 80% and significantly prolonged survival of treated mice in the orthotopic model. Taken together, our results suggest that RNAi therapeutics could effectively be delivered in a localized manner with HA-coated LNPs and ultimately may become a therapeutic modality for GBM.
多形性胶质母细胞瘤（GBM）是最具浸润性、侵袭性和治疗效果最差的脑肿瘤之一。基因组学和蛋白质组学的进展为确定治疗 GBM 的潜在治疗靶点铺平了道路，但这些治疗 GBM 的主要候选药物绝大多数都无效，主要是因为它们穿过血脑屏障的通道受限。由于纳米颗粒能够将药物输送到靶位，同时最小化不良反应，因此它们已成为治疗各种类型肿瘤的有前途的平台。在此，我们设计了一种局部策略，使用透明质酸（HA）接枝脂质纳米颗粒（LNPs）将 RNA 干扰（RNAi）直接递送到 GBM 部位。先前已经表明，具有离子化脂质的 LNPs 非常有效地将小干扰 RNA（siRNA）递送到各种细胞类型中。LNP 的表面用透明质酸（HA）功能化，透明质酸是一种天然存在的糖胺聚糖，专门与 GBM 细胞上表达的 CD44 受体结合。我们发现 HA-LNPs 可以成功地与 GBM 细胞系和 GBM 患者的原代神经球结合。载有 Polo-Like Kinase 1（PLK1）siRNA（siPLK1）的 HA-LNPs 可显著降低 PLK1 mRNA 的表达，并在剪切流下导致细胞死亡，剪切流模拟脑脊液的流动，与对照组相比。接下来，通过将 U87MG 细胞颅内注射到裸鼠中建立人 GBM U87MG 原位异种移植模型。进行了 HA-LNPs 包封的 Cy3-siRNA 的对流，并且在 U87MG 细胞中观察到特异性 Cy3 摄取。此外，HA-LNPs 包封的 siPLK1 对流使 mRNA 水平降低了 80%以上，并显著延长了原位模型中治疗小鼠的存活时间。总之，我们的结果表明，HA 涂层 LNPs 可以有效地以局部方式递送电镜 RNA 治疗药物，最终可能成为 GBM 的一种治疗方式。
