Institute of Pharmacology, National Yang Ming Chiao Tung University, Taipei, 112, Taiwan.
Faculty of Pharmacy, National Yang Ming Chiao Tung University, Taipei, 112, Taiwan.
Arch Pharm Res. 2024 Nov;47(10-11):829-853. doi: 10.1007/s12272-024-01514-0. Epub 2024 Nov 1.
Human antigen R (HuR), an RNA-binding protein, is implicated in regulating mRNA stability and translation in cancer, especially in triple-negative breast cancer (TNBC), a highly aggressive form. CRISPR/Cas9-mediated HuR knockout (HuR CRISPR) presents a promising genetic therapeutic approach, but it encounters transfection limitations. Docetaxel (DTX), an effective cytotoxic agent against metastatic breast cancer (BC), faces challenges related to vehicle-associated adverse events in DTX formulations. Therefore, we designed multifunctional nanoparticles with pH-sensitive PEG derivatives and targeting peptides to enable efficient HuR CRISPR and DTX delivery to human TNBC MDA-MB-231 cells and tumor-bearing mice. Our findings indicated that these nanoparticles displayed pH-responsive cytotoxicity, precise EGFR targeting, efficient tumor penetration, successful endosomal escape, and accurate nuclear and cytoplasmic localization. They also demonstrated the ability to spare normal cells and prevent hemolysis. Our study concurrently modulated multiple pathways, including EGFR, Wnt/β-catenin, MDR, and EMT, through the regulation of EGFR/PI3K/AKT, HuR/galectin-3/GSK-3β/β-catenin, and P-gp/MRPs/BCRP, as well as YAP1/TGF-β/ZEB1/Slug/MMPs. The combined treatment arrested the cell cycle at the G2 phase and inhibited EMT, effectively impeding tumor progression. Tissue distribution, biochemical assays, and histological staining revealed the enhanced safety profile of pH-responsive PEG- and peptide-modified nanoformulations in TNBC mice. The DTX-embedded and peptide-modified nanoparticles mitigated the side effects of DTX, enhanced cytotoxicity in TNBC MDA-MB-231 cells, and exhibited remarkable antitumor efficacy and safety in TNBC-bearing mice with HuR CRISPR deletion. Collectively, the combination therapy of DTX and CRISPR/Cas9 offers an effective platform for delivering antineoplastic agents and gene-editing systems to combat tumor resistance and progression in TNBC.
人抗原 R（HuR）是一种 RNA 结合蛋白，与癌症中的 mRNA 稳定性和翻译调节有关，特别是在三阴性乳腺癌（TNBC）中，这种调节作用更为显著。CRISPR/Cas9 介导的 HuR 敲除（HuR CRISPR）是一种很有前途的基因治疗方法，但它面临着转染的局限性。多西紫杉醇（DTX）是一种有效的转移性乳腺癌（BC）细胞毒药物，但在 DTX 制剂中存在与载体相关的不良反应的挑战。因此，我们设计了具有 pH 敏感性 PEG 衍生物和靶向肽的多功能纳米粒子，以实现有效的 HuR CRISPR 和 DTX 递送到人 TNBC MDA-MB-231 细胞和荷瘤小鼠。我们的研究结果表明，这些纳米粒子表现出 pH 响应性细胞毒性、精确的 EGFR 靶向、高效的肿瘤穿透、成功的内体逃逸以及准确的核质定位。它们还能够保护正常细胞并防止溶血。我们的研究同时通过调节 EGFR/PI3K/AKT、HuR/半乳糖凝集素-3/GSK-3β/β-catenin 和 P-gp/MRPs/BCRP 以及 YAP1/TGF-β/ZEB1/Slug/MMPs 等多种途径，同时调节了多个途径，包括 EGFR、Wnt/β-catenin、MDR 和 EMT。联合治疗将细胞周期阻滞在 G2 期，并抑制 EMT，有效阻止肿瘤进展。组织分布、生化测定和组织学染色显示，pH 响应性 PEG 和肽修饰的纳米制剂在 TNBC 小鼠中的安全性得到了提高。载有 DTX 的肽修饰纳米粒子减轻了 DTX 的副作用，增强了 TNBC MDA-MB-231 细胞的细胞毒性，并在 HuR CRISPR 缺失的 TNBC 荷瘤小鼠中表现出显著的抗肿瘤疗效和安全性。总的来说，DTX 和 CRISPR/Cas9 的联合治疗为递送抗肿瘤药物和基因编辑系统提供了一个有效的平台，以对抗 TNBC 中的肿瘤耐药性和进展。
