Institute of Pharmacology, National Yang-Ming University, Taipei 11221, Taiwan.
Nanoscale Horiz. 2021 Sep 1;6(9):729-743. doi: 10.1039/d1nh00254f. Epub 2021 Jul 29.
Head and neck cancer (HNC) has a high incidence and a poor prognosis. Epirubicin, a topoisomerase inhibitor, is a potential anthracycline chemotherapeutic for HNC treatment. HuR (ELAVL1), an RNA-binding protein, plays a critical role in promoting tumor survival, invasion, and resistance. HuR knockout via CRISPR/Cas9 (HuR CRISPR) is a possible strategy for the simultaneous modulation of the various pathways of tumor progression. Multifunctional nanoparticles modified with pH-sensitive epidermal growth factor receptor (EGFR)-targeting and nucleus-directed peptides were designed for the efficient delivery of HuR CRISPR and epirubicin to human tongue squamous carcinoma SAS cells and SAS tumor-bearing mice. The pH-sensitive nanoparticles responded to the acidic pH value as a switch to expose the targeting peptides. The cellular uptake and transfection efficiency of these nanoparticles in SAS cells increased via EGFR targeting, ligand-mediated endocytosis, and endosomal escape. These nanoparticles showed low cytotoxicity towards normal oral keratinocyte NOK cells. CRISPR/Cas9 was transported into the nucleus via the nuclear directing peptide and successfully knocked out HuR to suppress proliferation, metastasis, and resistance in SAS cells. The multiple inhibition of EGFR/β-catenin/epithelial-mesenchymal transition pathways was mediated through modulating the EGFR/PI3K/mTOR/AKT axis. The co-treatment of epirubicin and HuR CRISPR in SAS cells further facilitated apoptosis/necroptosis/autophagy and caused cancer cell death. In combination with HuR CRISPR nanoparticles, the efficacy and safety of epirubicin nanoparticles against cancer in SAS tumor-bearing mice improved significantly. Collectively, these nanoparticles showed a tumor pH response, active EGFR targeting, and nuclear localization and thus offered a combinatorial spatiotemporal platform for chemotherapy and the CRISPR/Cas gene-editing system.
头颈部癌症（HNC）发病率高，预后差。表阿霉素是一种拓扑异构酶抑制剂，是治疗 HNC 的潜在蒽环类化疗药物。HuR（ELAVL1）是一种 RNA 结合蛋白，在促进肿瘤存活、侵袭和耐药性方面发挥着关键作用。通过 CRISPR/Cas9（HuR CRISPR）敲除 HuR 是同时调节肿瘤进展的各种途径的一种可能策略。设计了具有 pH 敏感表皮生长因子受体（EGFR）靶向和核靶向肽的多功能纳米粒子，用于有效递送至人舌鳞癌细胞 SAS 细胞和 SAS 荷瘤小鼠的 HuR CRISPR 和表阿霉素。这些 pH 敏感的纳米粒子响应酸性 pH 值作为开关，暴露靶向肽。通过 EGFR 靶向、配体介导的内吞作用和内涵体逃逸，增加了这些纳米粒子在 SAS 细胞中的细胞摄取和转染效率。这些纳米粒子对正常口腔角质形成细胞 NOK 细胞的细胞毒性低。CRISPR/Cas9 通过核靶向肽进入细胞核，并成功敲除 HuR，以抑制 SAS 细胞的增殖、转移和耐药性。通过调节 EGFR/PI3K/mTOR/AKT 轴，介导了对 EGFR/β-catenin/上皮间质转化途径的多重抑制。在 SAS 细胞中联合使用表阿霉素和 HuR CRISPR 进一步促进了细胞凋亡/坏死/自噬，并导致癌细胞死亡。与 HuR CRISPR 纳米粒子联合使用时，显著提高了载药纳米粒子对 SAS 荷瘤小鼠癌症的疗效和安全性。总之，这些纳米粒子表现出肿瘤 pH 响应、主动 EGFR 靶向和核定位，从而提供了化疗和 CRISPR/Cas 基因编辑系统的组合时空平台。
