National Center for International Research of Biotargeting Theranostics, Guangxi Key Laboratory of Biotargeting Theranostics, Guangxi Medical University, Nanning, Guangxi, 530021, China.
Department of Hepatobiliary Surgery, The First Affiliated Hospital, Guangxi Medical University, Nanning, Guangxi, 530021, China.
Signal Transduct Target Ther. 2021 Dec 3;6(1):411. doi: 10.1038/s41392-021-00799-7.
Non-human primates (NHPs) represent the most valuable animals for drug discovery. However, the current main challenge remains that the NHP has not yet been used to develop an efficient translational medicine platform simulating human diseases, such as cancer. This study generated an in situ gene-editing approach to induce efficient loss-of-function mutations of Pten and p53 genes for rapid modeling primary and metastatic liver tumors using the CRISPR/Cas9 in the adult cynomolgus monkey. Under ultrasound guidance, the CRISPR/Cas9 was injected into the cynomolgus monkey liver through the intrahepatic portal vein. The results showed that the ultrasound-guided CRISPR/Cas9 resulted in indels of the Pten and p53 genes in seven out of eight monkeys. The best mutation efficiencies for Pten and p53 were up to 74.71% and 74.68%, respectively. Furthermore, the morbidity of primary and extensively metastatic (lung, spleen, lymph nodes) hepatoma in CRISPR-treated monkeys was 87.5%. The ultrasound-guided CRISPR system could have great potential to successfully pursue the desired target genes, thereby reducing possible side effects associated with hitting non-specific off-target genes, and significantly increasing more efficiency as well as higher specificity of in situ gene editing in vivo, which holds promise as a powerful, yet feasible tool, to edit disease genes to build corresponding human disease models in adult NHPs and to greatly accelerate the discovery of new drugs and save economic costs.
非人类灵长类动物（NHPs）是药物发现最有价值的动物。然而，目前的主要挑战仍然是，NHP 尚未被用于开发模拟人类疾病（如癌症）的高效转化医学平台。本研究采用 CRISPR/Cas9 在成年食蟹猴中诱导 Pten 和 p53 基因的高效功能丧失突变，生成一种原位基因编辑方法，用于快速建模原发性和转移性肝肿瘤。在超声引导下，将 CRISPR/Cas9 通过肝内门静脉注入食蟹猴肝脏。结果表明，在 8 只猴子中有 7 只猴子的 CRISPR/Cas9 导致 Pten 和 p53 基因发生插入缺失。Pten 和 p53 的最佳突变效率分别高达 74.71%和 74.68%。此外，CRISPR 处理猴子原发性和广泛转移性（肺、脾、淋巴结）肝癌的发病率为 87.5%。超声引导的 CRISPR 系统具有很大的潜力，可以成功地追求所需的靶基因，从而减少与击中非特异性脱靶基因相关的可能副作用，并显著提高体内原位基因编辑的效率和更高的特异性，有望成为一种强大且可行的工具，用于编辑疾病基因，构建相应的人类疾病模型，从而大大加快新药的发现并节省经济成本。
