Generation of porcine hepatocellular carcinoma model by P53 modification and aflatoxin B1 intake in Bamaxiang pigs. Bacteria

Condons 248 and 249 have been identified as two of the few mutational major hotspots of human P53, and Aflatoxin B1 (AFB1) is hepatotoxic. In this study, we firstly obtained F0 P53 gene-modified Bamaxiang boars carrying 241W and 242S mutations (corresponding to 248W and 249S mutations in human P53) via CRISPR/Cas9 and somatic cell nuclear tranfer (SCNT) techniques, and F2 pigs were generated by subsequent breeding. After feeding AFB1 for several months, the F0 and F2 homozygous gene-edited pigs started to exhibited distinct anorexia, depression and growth retardation symptoms along with dramatically increased serum alpha-fetoprotein (AFP) levels. Pathological section and immunohistochemistry staining analyses demonstrated that all P53 gene-edited Bamaxiang pigs manifested hepatocellular carcinoma (HCC) phenotype after AFB1-induced for 2-6 months, but no tumor was found in other tissues. Proteomic analysis identified 1034 DEPs between the 241W242S and WT pigs (both fed with AFB1), and two-thirds of the top 50 DEPs were reported involving HCC. Metagenomics sequencing revealed significant differences in faecal microbiota composition between the genetically modified and WT pigs, both fed with AFB1. Our results demonstrate that we have established an inducible porcine HCC model, which is highly practical and can be very useful in studying human HCC