Elkhadragy Lobna, Carlino Maximillian J, Jordan Luke R, Pennix Thomas, Ismail Nahed, Guzman Grace, Samuelson Jonathan P, Schook Lawrence B, Schachtschneider Kyle M, Gaba Ron C
Department of Radiology, University of Illinois at Chicago, Chicago, IL 60612, USA.
Department of Pathology, University of Illinois at Chicago, Chicago, IL 60612, USA.
Dis Model Mech. 2025 Jan 1;18(1). doi: 10.1242/dmm.052079. Epub 2025 Jan 29.
Hepatocellular carcinoma (HCC) is an aggressive disease with poor prognosis, necessitating preclinical models for evaluating novel therapies. Large-animal models are particularly valuable for assessing locoregional therapies, which are widely employed across HCC stages. This study aimed to develop a large-animal HCC model with tailored tumor mutations. The Oncopig, a genetically engineered pig with inducible TP53R167H and KRASG12D, was used in the study. Hepatocytes were isolated from Oncopigs and exposed to Cre recombinase in vitro to create HCC cells, and additional mutations were introduced by CRISPR/Cas9 knockout of PTEN and CDKN2A. These edits increased Oncopig HCC cell proliferation and migration. Autologous HCC cells with these CRISPR edits were implanted into Oncopigs using two approaches: ultrasound-guided percutaneous liver injections, which resulted in the development of localized intrahepatic masses, and portal vein injections, which led to multifocal tumors that regressed over time. Tumors developed by both approaches harbored PTEN and CDKN2A knockout mutations. This study demonstrates the feasibility of developing genetically tailored HCC tumors in Oncopigs using somatic cell CRISPR editing and autologous implantation, providing a valuable large-animal model for in vivo therapeutic assessment.
肝细胞癌（HCC）是一种侵袭性疾病，预后较差，因此需要临床前模型来评估新疗法。大型动物模型对于评估局部区域疗法特别有价值，这种疗法在HCC的各个阶段都被广泛应用。本研究旨在开发一种具有定制肿瘤突变的大型动物HCC模型。研究中使用了一种基因工程猪Oncopig，其具有可诱导的TP53R167H和KRASG12D。从Oncopig中分离出肝细胞，并在体外将其暴露于Cre重组酶以产生HCC细胞，通过CRISPR/Cas9敲除PTEN和CDKN2A引入额外的突变。这些编辑增加了Oncopig HCC细胞的增殖和迁移。采用两种方法将具有这些CRISPR编辑的自体HCC细胞植入Oncopig：超声引导下经皮肝注射，导致局部肝内肿块的形成；门静脉注射，导致多灶性肿瘤，随着时间推移会消退。两种方法产生的肿瘤都存在PTEN和CDKN2A敲除突变。本研究证明了使用体细胞CRISPR编辑和自体植入在Oncopig中开发基因定制的HCC肿瘤的可行性，为体内治疗评估提供了一个有价值的大型动物模型。
