Osteosarcoma (OS) is a highly aggressive pediatric cancer, characterized by frequent lung metastasis and pathologic bone destruction. Vascular endothelial growth factor A (VEGFA), highly expressed in OS, not only contributes to angiogenesis within the tumor microenvironment via paracrine stimulation of vascular endothelial cells, but also acts as an autocrine survival factor for tumor cell themselves, thus making it a promising therapeutic target for OS. CRISPR/Cas9 is a versatile genome editing technology and holds tremendous promise for cancer treatment. However, a major bottleneck to achieve the therapeutic potential of the CRISPR/Cas9 is the lack of in vivo tumor-targeted delivery systems. Here, we screened an OS cell-specific aptamer (LC09) and developed a LC09-functionalized PEG-PEI-Cholesterol (PPC) lipopolymer encapsulating CRISPR/Cas9 plasmids encoding VEGFA gRNA and Cas9. Our results demonstrated that LC09 facilitated selective distribution of CRISPR/Cas9 in both orthotopic OS and lung metastasis, leading to effective VEGFA genome editing in tumor, decreased VEGFA expression and secretion, inhibited orthotopic OS malignancy and lung metastasis, as well as reduced angiogenesis and bone lesion with no detectable toxicity. The delivery system simultaneously restrained autocrine and paracrine VEGFA signaling in tumor cells and could facilitate translating CRISPR-Cas9 into clinical cancer treatment.

中文翻译：

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通过适体功能化脂聚合物靶向递送 CRISPR/Cas9，用于骨肉瘤中 VEGFA 的治疗性基因组编辑


骨肉瘤（OS）是一种高度侵袭性的儿童癌症，其特征是频繁的肺转移和病理性骨破坏。血管内皮生长因子 A (VEGFA) 在 OS 中高表达，不仅通过旁分泌刺激血管内皮细胞促进肿瘤微环境中的血管生成，而且还充当肿瘤细胞本身的自分泌生存因子，从而使其成为一种有前途的治疗方法操作系统的目标。 CRISPR/Cas9 是一种多功能的基因组编辑技术，为癌症治疗带来了巨大的前景。然而，实现 CRISPR/Cas9 治疗潜力的一个主要瓶颈是缺乏体内肿瘤靶向递送系统。在这里，我们筛选了 OS 细胞特异性适体 (LC09)，并开发了一种 LC09 功能化的 PEG-PEI-胆固醇 (PPC) 脂聚合物，封装编码VEGFA gRNA 和 Cas9 的 CRISPR/Cas9 质粒。我们的结果表明，LC09促进了CRISPR/Cas9在原位OS和肺转移中的选择性分布，导致肿瘤中有效的VEGFA基因组编辑，减少了VEGFA的表达和分泌，抑制了原位OS恶性肿瘤和肺转移，并减少了血管生成和骨转移。没有可检测到的毒性的损伤。该递送系统同时抑制肿瘤细胞中的自分泌和旁分泌 VEGFA 信号传导，并有助于将 CRISPR-Cas9 转化为临床癌症治疗。