An arrhythmogenic cardiomyopathy pig model by PKP2 heterozygous knockout

Arrhythmogenic cardiomyopathy (ACM) patients are at higher risk of ventricular arrhythmias and sudden death. The inability to obtain samples of early lesions of ACM patients and the lack of suitable animal models limit the progress of ACM research. The characteristics of clinical, pathological and mutation loci of 24 ACM patients with PKP2 mutation were summarized. Patient carrying the c.1132C>T mutation, exhibited the earliest age of onset and presented both major adverse cardiovascular events and cardiac structural abnormalities. Additionally, we selected the previously reported PKP2 C>T pathogenic mutation (c.1147C>T), located near c.1132 of PKP2, to enhance the likelihood of successfully constructing the PKP2 mutant pig model. The BE3 gene editing system were used to induce C>T mutation, sgRNA1 and sgRNA2 in the PKP2 gene was designed to induce the c.1132C>T and c.1147C>T mutation. Target editing efficiencies of sgRNA1 and sgRNA2 were 42.9% and 25.9%, respectively. SgRNA1 were finally selected to construct PKP2+/- porcine fetal fibroblasts. 11 live and 3 dead PKP2+/- piglets were obtained. At 24-month-old, PKP2+/- pigs developed premature ventricular contraction and right ventricular dilatation. Pathological examination showed adipocytes infiltration was present in RV of PKP2+/- pigs. PKP2 haploinsufficiency resulted in reduced length and electron density of desmosomes. Transcriptome sequencing revealed that PKP2+/- pigs exhibited high expression of genes associated with lipid catabolic reaction. This study firstly constructed the PKP2+/- pigs model by used the BE3 gene-edit system.