Department of Biomedical Engineering, Tufts University, Medford, MA 02155.
Broad Institute of MIT and Harvard, Cambridge, MA 02142.
Proc Natl Acad Sci U S A. 2021 Mar 9;118(10). doi: 10.1073/pnas.2020401118.
Loss-of-function mutations in Angiopoietin-like 3 () are associated with lowered blood lipid levels, making an attractive therapeutic target for the treatment of human lipoprotein metabolism disorders. In this study, we developed a lipid nanoparticle delivery platform carrying Cas9 messenger RNA (mRNA) and guide RNA for CRISPR-Cas9-based genome editing of in vivo. This system mediated specific and efficient gene knockdown in the liver of wild-type C57BL/6 mice, resulting in profound reductions in serum ANGPTL3 protein, low density lipoprotein cholesterol, and triglyceride levels. Our delivery platform is significantly more efficient than the FDA-approved MC-3 LNP, the current gold standard. No evidence of off-target mutagenesis was detected at any of the nine top-predicted sites, and no evidence of toxicity was detected in the liver. Importantly, the therapeutic effect of genome editing was stable for at least 100 d after a single dose administration. This study highlights the potential of LNP-mediated delivery as a specific, effective, and safe platform for Cas9-based therapeutics.
ANGPTL3 基因的功能丧失性突变与降低血脂水平有关，使其成为治疗人类脂蛋白代谢紊乱的有吸引力的治疗靶点。在这项研究中，我们开发了一种携带 Cas9 信使 RNA（mRNA）和指导 RNA 的脂质纳米颗粒（LNP）递送平台，用于 CRISPR-Cas9 介导的体内基因编辑。该系统介导了野生型 C57BL/6 小鼠肝脏中特定和有效的基因敲低，导致血清 ANGPTL3 蛋白、低密度脂蛋白胆固醇和甘油三酯水平的显著降低。我们的递送平台比 FDA 批准的 MC-3 LNP（目前的金标准）效率更高。在九个预测的靶位点中，没有检测到任何脱靶突变的证据，在肝脏中也没有检测到毒性的证据。重要的是，单次给药后，基因编辑的治疗效果至少稳定 100 天。这项研究强调了 LNP 介导的递送作为 Cas9 治疗的一种特异性、有效性和安全性平台的潜力。
