Agbaria Majd, Jbara-Agbaria Doaa, Grad Etty, Ben-David-Naim Meital, Aizik Gil, Golomb Gershon
Institute for Drug Research, School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem 9112001, Israel.
Institute for Drug Research, School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem 9112001, Israel; The Center for Nanoscience and Nanotechnology, The Hebrew University of Jerusalem, Jerusalem 9190401, Israel.
J Control Release. 2023 Mar;355:312-326. doi: 10.1016/j.jconrel.2023.01.084. Epub 2023 Feb 9.
Pancreatic ductal adenocarcinoma (PDAC) is among the leading causes of cancer-related death, and it is highly resistant to therapy owing to its unique extracellular matrix. VAV1 protein, overexpressed in several cancer diseases including pancreatic cancer (PC), increases tumor proliferation and enhances metastases formation, which are associated with decreased survival. We hypothesized that an additive anti-tumor effect could be obtained by co-encapsulating in PLGA nanoparticles (NPs), the negatively charged siRNA against VAV1 (siVAV1) with the positively charged anti-tumor LL37 peptide, as a counter-ion. Several types of NPs were formulated and were characterized for their physicochemical properties, cellular internalization, and bioactivity in vitro. NPs' biodistribution, toxicity, and bioactivity were examined in a mice PDAC model. An optimal siVAV1 formulation (siVAV1-LL37 NPs) was characterized with desirable physicochemical properties in terms of nano-size, low polydispersity index (PDI), neutral surface charge, high siVAV1 encapsulation efficiency, spherical shape, and long-term shelf-life stability. Cell assays demonstrated rapid engulfment by PC cells, a specific and significant dose-dependent proliferation inhibition, as well as knockdown of VAV1 mRNA levels and migration inhibition in VAV1 cells. Treatment with siVAV1-LL37 NPs in the mice PDAC model revealed marked accumulation of NPs in the liver and in the tumor, resulting in an increased survival rate following suppression of tumor growth and metastases, mediated via the knockdown of both VAV1 mRNA and protein levels. This proof-of-concept study validates our hypothesis of an additive effect in the treatment of PC facilitated by co-encapsulating siVAV1 in NPs with LL37 serving a dual role as a counter ion as well as an anti-tumor agent.
胰腺导管腺癌（PDAC）是癌症相关死亡的主要原因之一，由于其独特的细胞外基质，它对治疗具有高度抗性。VAV1蛋白在包括胰腺癌（PC）在内的几种癌症疾病中过度表达，会增加肿瘤增殖并促进转移形成，这与生存率降低有关。我们假设，通过将带负电荷的针对VAV1的小干扰RNA（siVAV1）与带正电荷的抗肿瘤LL37肽作为抗衡离子共包封在聚乳酸-羟基乙酸共聚物纳米颗粒（NPs）中，可以获得相加的抗肿瘤效果。制备了几种类型的纳米颗粒，并对其理化性质、细胞内化和体外生物活性进行了表征。在小鼠PDAC模型中检测了纳米颗粒的生物分布、毒性和生物活性。一种最佳的siVAV1制剂（siVAV1-LL37 NPs）具有理想的理化性质，包括纳米尺寸、低多分散指数（PDI）、中性表面电荷、高siVAV1包封效率、球形形状和长期储存稳定性。细胞实验表明，PC细胞能快速摄取，具有特异性且显著的剂量依赖性增殖抑制作用，以及VAV1细胞中VAV1 mRNA水平的敲低和迁移抑制。在小鼠PDAC模型中用siVAV1-LL37 NPs治疗显示，纳米颗粒在肝脏和肿瘤中显著蓄积，通过敲低VAV1 mRNA和蛋白水平，抑制肿瘤生长和转移后，生存率提高。这项概念验证研究证实了我们的假设，即通过将siVAV1与LL37共包封在纳米颗粒中治疗PC具有相加效应，LL37兼具抗衡离子和抗肿瘤剂的双重作用。
