Department of Viral Immunology, Scripps Korea Antibody Institute, Chuncheon, South Korea.
Center for Theragnosis, Biomedical Research Institute, Korea Institute of Science and Technology, Seoul, South Korea.
Biomaterials. 2021 May;272:120793. doi: 10.1016/j.biomaterials.2021.120793. Epub 2021 Apr 1.
Therapeutic application of CRISPR/Cas9 nucleases remains a challenge due to the lack of efficient in vivo delivery carriers. Here, we examine the ability of lentiviral vectors pseudotyped with hepatitis C virus (HCV)/E1E2 envelope glycoproteins to systemically deliver CRISPR/Cas9 to hepatic tumors in vivo. We demonstrated that systemic administration of E1E2-pseudotyped lentiviral vectors can selectively deliver Cas9 and sgRNA specific for kinesin spindle protein (KSP) to Huh7 tumors in the orthotopic Huh7 mice due to the specific interactions between E1E2 and their cellular receptors. This specific delivery leads to effective KSP gene disruption, potently inhibiting tumor growth. Furthermore, we demonstrated that E1E2-pseudotyping is more suitable for systemic delivery of CRISPR/Cas9 in cancer therapy than vesicular stomatitis virus-pseudotyping, the most widely used pseudotyping, because of stability in human serum, little transduction to DCs, low innate immune response, and cell-specific targeting ability. This study suggests that E1E2-pseudotyped lentivirus carrying CRISPR/Cas9 can substantially benefit the treatment of Huh7 tumors.
CRISPR/Cas9 核酸酶的治疗应用仍然是一个挑战，因为缺乏有效的体内递送载体。在这里，我们研究了丙型肝炎病毒 (HCV)/E1E2 包膜糖蛋白假型慢病毒载体将 CRISPR/Cas9 系统递送至体内肝肿瘤的能力。我们证明，由于 E1E2 与其细胞受体之间的特异性相互作用，系统给予 E1E2 假型慢病毒载体可以选择性地将 Cas9 和 sgRNA 递送至原位 Huh7 小鼠中的 Huh7 肿瘤，该 sgRNA 特异性针对驱动蛋白纺锤体蛋白 (KSP)。这种特异性递送导致有效的 KSP 基因破坏，强力抑制肿瘤生长。此外，我们证明，E1E2 假型化比最广泛使用的假型化——水疱性口炎病毒假型化更适合用于癌症治疗的 CRISPR/Cas9 的系统递送，因为其在人血清中稳定、向树突状细胞的转导很少、低固有免疫反应和细胞特异性靶向能力。这项研究表明，携带 CRISPR/Cas9 的 E1E2 假型慢病毒可以极大地有益于 Huh7 肿瘤的治疗。
