Generation of in situ CRISPR-mediated primary and metastatic cancer from monkey liver

Abstract

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.

Emerging Nano-/Biotechnology Drives Oncolytic Virus-Activated and Combined Cancer Immunotherapy

“…Unfortunately, the absence of spontaneous orthotopic tumor model hinders this progression of new drugs. Zhong and coworkers [ 151 ] evaluated ultrasound-guided infusion of CRISPR-Cas9 into the liver of cynomolgus monkeys via the intrahepatic portal vein. They demonstrated that Pten and p53 gene mutations could be efficiently generated in the liver of these monkeys through intrahepatic portal vein injection of the CRISPR-Cas9 genome editing system.…”

Gene editing monkeys: Retrospect and outlook

“…As expected, the model monkey died shortly after birth and displayed severe developmental retardation. A recent study reported a new method to generate gene-modified monkeys by in situ CRISPR-mediated technique ( Zhong et al, 2021 ). This study utilized CRISPR/Cas9 to knock down Pten and p53 genes in adult cynomolgus, thus modeling primary and metastatic liver tumors rapidly, which was effective in situ gene editing approach.…”

Unexpected mutations occurred in CRISPR/Cas9 edited Drosophila analyzed by deeply whole genomic sequencing

“…Due to the characteristics of simple composition and high editing efficiency [ [4] , [5] , [6] ], it has rapidly gained popularity and become as a hallmark in biomedicine field. CRISPR/Cas9 can be universally applied to a wide range of biological studies, including nonhuman primates [ 7 ], mice [ 8 ], rat [ 9 ], zebra fish [ 10 ], Drosophila [ 11 ], helminth [ 12 ], as well as a myriad of common plants [ 13 ] and microbes [ 14 ]. CRISPR/Cas9 gene editing has found widespread application, however, there remains no definitive conclusion regarding whether the process of gene editing and repair can potentially induce harmful effects on the genome.…”