Department of Pathology, University of Pittsburg School of Medicine, Pittsburgh, Pennsylvania, USA.
Pittsburgh Liver Research Center at Pittsburgh Liver Institute, Animal Imaging Center, University of Pittsburg School of Medicine, Pittsburgh, Pennsylvania, USA.
Hepatol Commun. 2024 Mar 18;8(4). doi: 10.1097/HC9.0000000000000412. eCollection 2024 Apr 1.
Liver cancer is one of the most lethal malignancies for humans. The treatment options for advanced-stage liver cancer remain limited. A new treatment is urgently needed to reduce the mortality of the disease.
In this report, we developed a technology for mutation site insertion of a suicide gene (herpes simplex virus type 1- thymidine kinase) based on type II CRISPR RNA-guided endonuclease Cas9-mediated genome editing to treat liver cancers.
We applied the strategy to 3 different mutations: S45P mutation of catenin beta 1, chromosome breakpoint of solute carrier family 45 member 2-alpha-methylacyl-CoA racemase gene fusion, and V235G mutation of SAFB-like transcription modulator. The results showed that the herpes simplex virus type 1-thymidine kinase insertion rate at the S45P mutation site of catenin beta 1 reached 77.8%, while the insertion rates at the breakpoint of solute carrier family 45 member 2 - alpha-methylacyl-CoA racemase gene fusion were 95.1%-98.7%, and the insertion at V235G of SAFB-like transcription modulator was 51.4%. When these targeting reagents were applied to treat mouse spontaneous liver cancer induced by catenin beta 1S45P or solute carrier family 45 member 2-alpha-methylacyl-CoA racemase, the mice experienced reduced tumor burden and increased survival rate. Similar results were also obtained for the xenografted liver cancer model: Significant reduction of tumor volume, reduction of metastasis rate, and improved survival were found in mice treated with the targeting reagent, in comparison with the control-treated groups.
Our studies suggested that mutation targeting may hold promise as a versatile and effective approach to treating liver cancers.
肝癌是人类最致命的恶性肿瘤之一。晚期肝癌的治疗选择仍然有限。迫切需要新的治疗方法来降低疾病死亡率。
在本报告中，我们开发了一种基于 II 型 CRISPR RNA 指导的内切酶 Cas9 介导的基因组编辑的自杀基因（单纯疱疹病毒 1-胸苷激酶）突变位点插入技术，用于治疗肝癌。
我们将该策略应用于 3 种不同的突变：连接蛋白 beta 1 的 S45P 突变、溶质载体家族 45 成员 2- -甲基酰基辅酶 A 消旋酶基因融合的染色体断点和 SAFB 样转录调节剂的 V235G 突变。结果表明，连接蛋白 beta 1 的 S45P 突变位点的单纯疱疹病毒 1-胸苷激酶插入率达到 77.8%，而溶质载体家族 45 成员 2 - -甲基酰基辅酶 A 消旋酶基因融合的插入率为 95.1%-98.7%，SAFB 样转录调节剂的 V235G 插入率为 51.4%。当这些靶向试剂用于治疗连接蛋白 beta 1S45P 或溶质载体家族 45 成员 2- -甲基酰基辅酶 A 消旋酶诱导的小鼠自发性肝癌时，小鼠的肿瘤负担减少，存活率提高。在异种移植肝癌模型中也得到了类似的结果：与对照组相比，用靶向试剂治疗的小鼠肿瘤体积显著减小，转移率降低，存活率提高。
我们的研究表明，突变靶向可能是治疗肝癌的一种有前途的通用有效方法。
