Harnessing CRISPR-Cas9 technology for cancer therapeutics has been hampered by low editing efficiency in tumors and potential toxicity of existing delivery systems. Here, we describe a safe and efficient lipid nanoparticle (LNP) for the delivery of Cas9 mRNA and sgRNAs that use a novel amino-ionizable lipid. A single intracerebral injection of CRISPR-LNPs against PLK1 (sgPLK1-cLNPs) into aggressive orthotopic glioblastoma enabled up to ~70% gene editing in vivo, which caused tumor cell apoptosis, inhibited tumor growth by 50%, and improved survival by 30%. To reach disseminated tumors, cLNPs were also engineered for antibody-targeted delivery. Intraperitoneal injections of EGFR-targeted sgPLK1-cLNPs caused their selective uptake into disseminated ovarian tumors, enabled up to ~80% gene editing in vivo, inhibited tumor growth, and increased survival by 80%. The ability to disrupt gene expression in vivo in tumors opens new avenues for cancer treatment and research and potential applications for targeted gene editing of noncancerous tissues.

由于肿瘤编辑效率低和现有递送系统的潜在毒性，阻碍了将 CRISPR-Cas9 技术用于癌症治疗。在这里，我们描述了一种安全有效的脂质纳米颗粒 (LNP)，用于递送使用新型氨基可电离脂质的 Cas9 mRNA 和 sgRNA。针对PLK1的单次脑内注射 CRISPR-LNP(sgPLK1-cLNPs) 进入侵袭性原位胶质母细胞瘤，可在体内实现高达约 70% 的基因编辑，从而导致肿瘤细胞凋亡，抑制肿瘤生长 50%，并将存活率提高 30%。为了到达播散性肿瘤，cLNPs 也被设计用于抗体靶向递送。腹腔内注射靶向 EGFR 的 sgPLK1-cLNP 可使其选择性吸收到播散性卵巢肿瘤中，从而实现高达约 80% 的体内基因编辑，抑制肿瘤生长，并将存活率提高 80%。破坏肿瘤体内基因表达的能力为癌症治疗和研究以及非癌组织靶向基因编辑的潜在应用开辟了新途径。