School of Chemistry, University of New South Wales, Sydney, 2052, Australia.
School of Women's and Children's Health, Faculty of Medicine and Health, University of New South Wales, Australia.
J Mater Chem B. 2021 Nov 17;9(44):9123-9135. doi: 10.1039/d1tb01837j.
High-grade serous ovarian cancer (HGSOC) is the most lethal gynaecological malignancy. Most patients are diagnosed at late stages when the tumour has metastasised throughout the peritoneal cavity. The Wnt receptor ROR2 has been identified as a promising therapeutic target in HGSOC, with limited targeting therapeutic options currently available. Small interfering RNA (siRNA)-based therapeutics hold great potential for inhibiting the function of specific biomarkers, however major challenges remain in efficient delivery and stability. The aim of this study was to investigate the ability of nanoparticles to deliver ROR2 siRNA into HGSOC cells, including platinum resistant models, and estimate the anti-metastatic effect a 3D organotypic model for ovarian cancer. The nanoparticles were generated by conjugating poly[2-(dimethylamino) ethyl methacrylate] (PDMAEMA) of various chain length to bovine serum albumin (BSA), followed by the condensation of ROR2 siRNA into polyplexes, also termed polyion complex (PIC) nanoparticles. The toxicity and uptake of ROR2 siRNA PIC nanoparticles in two HGSOC cell lines, CaOV3 as well as its cisplatin resistant pair (CaOV3CisR), in addition to primary cells used for the 3D organotypic model were investigated. ROR2 knockdown at both transcriptional and translational levels were evaluated real-time PCR and western blot analysis, respectively. Following 24 h incubation with the nanoparticles, functional assays were performed including proliferation (IncuCyte S3), transwell migration and 3D co-cultured transwell invasion assays. The PICs nanoparticles exhibited negligible toxicity in the paired CaOV3 cell lines or primary cells. Treating CaOV3 and CaOV3CisR cells with ROR2 siRNA containing PICs nanoparticles significantly inhibited migration and invasion ability. The biocompatible ROR2 siRNA conjugated PICs nanoparticles provide an innovative therapeutic option. ROR2 targeting therapy shows potential in treating HGSOC including platinum resistant forms.
高级别浆液性卵巢癌（HGSOC）是最致命的妇科恶性肿瘤。大多数患者在肿瘤已转移至整个腹腔的晚期才被诊断出来。Wnt 受体 ROR2 已被确定为 HGSOC 中有前途的治疗靶点，目前可用的靶向治疗选择有限。基于小干扰 RNA（siRNA）的治疗方法在抑制特定生物标志物的功能方面具有很大的潜力，然而，在有效递送上仍然存在重大挑战。本研究旨在研究纳米颗粒将 ROR2 siRNA 递送至包括铂耐药模型在内的 HGSOC 细胞的能力，并估计其在卵巢癌 3D 器官型模型中的抗转移作用。纳米颗粒通过将聚[2-（二甲氨基）乙基甲基丙烯酸酯]（PDMAEMA）与牛血清白蛋白（BSA）偶联，然后将 ROR2 siRNA 缩合成交联复合物，也称为聚离子复合物（PIC）纳米颗粒来生成。研究了两种 HGSOC 细胞系（CaOV3 及其顺铂耐药对（CaOV3CisR））以及用于 3D 器官型模型的原代细胞中 ROR2 siRNA PIC 纳米颗粒的毒性和摄取。通过实时 PCR 和 Western blot 分析分别评估了转录和翻译水平的 ROR2 敲低。在用纳米颗粒孵育 24 小时后，进行了功能测定，包括增殖（IncuCyte S3）、Transwell 迁移和 3D 共培养 Transwell 侵袭测定。在配对的 CaOV3 细胞系或原代细胞中，PIC 纳米颗粒表现出可忽略的毒性。用含 ROR2 siRNA 的 PIC 纳米颗粒处理 CaOV3 和 CaOV3CisR 细胞显著抑制了迁移和侵袭能力。ROR2 siRNA 缀合的 PIC 纳米颗粒具有生物相容性，为治疗 HGSOC 提供了一种创新的治疗选择。ROR2 靶向治疗在治疗包括铂耐药形式在内的 HGSOC 方面具有潜力。
