Disruption of anthrax toxin receptor 1 in pigs leads to a rare disease phenotype and protection from senecavirus A infection

Abstract

Senecavirus A (SVA) is a cause of vesicular disease in pigs, and infection rates are rising within the swine industry. Recently, anthrax toxin receptor 1 (ANTXR1) was revealed as the receptor for SVA in human cells. Herein, the role of ANTXR1 as a receptor for SVA in pigs was investigated by CRISPR/Cas9 genome editing. Strikingly, ANTXR1 knockout (KO) pigs exhibited features consistent with the rare disease, GAPO syndrome, in humans. Fibroblasts from wild type (WT) pigs supported replication of SVA; whereas, fibroblasts from KO pigs were resistant to infection. During an SVA challenge, clinical symptoms, including vesicular lesions, and circulating viremia were present in infected WT pigs but were absent in KO pigs. Additional ANTXR1-edited piglets were generated that were homozygous for an in-frame (IF) mutation. While IF pigs presented a GAPO phenotype similar to the KO pigs, fibroblasts showed mild infection, and circulating SVA nucleic acid was decreased in IF compared to WT pigs. Thus, this new ANTXR1 mutation resulted in decreased permissiveness of SVA in pigs. Overall, genetic disruption of ANTXR1 in pigs provides a unique model for GAPO syndrome and prevents circulating SVA infection and clinical symptoms, confirming that ANTXR1 acts as a receptor for the virus.

Porcine ANTXR1, Heparan Sulfate and Neu5Gc act as entry factors for Seneca Valley virus invasion

“…Mutations in ANTXR1 cause progressive extracellular-matrix accumulation in patients with the GAPO syndrome, a complex phenotype consisting of growth retardation, alopecia, pseudoanodontia and progressive optic atrophy 28 . Recently, ANTXR1 knockout pigs were produced, and as expected, exhibited resistance to SVV infection, while these pigs also developed GAPO-like symptoms 29 . Based on the above results, more explicit structural investigations on the SVV-ANTXR1 complex are warranted in order to design accurate editing sites that destroy the function of virus receptor while maintaining its normal physiological function 30 .…”

Gene editing of pigs to control influenza A virus infections

“…The best example of gene-edited, disease-resistant pigs is the generation of CD163 KO pigs, which have been shown to be fully resistant to porcine reproductive and respiratory syndrome virus infections [27][28][29] . Other examples include the disruption of the porcine amino peptidase N (APN) or the anthrax toxin receptor 1 (ANTXR1) which resulted in resistance to infection with transmissible gastroenteritis virus or senecavirus A virus, respectively 30,31 . In the present study, a stable colony of homozygous GE TMPRSS2 KO pigs was established.…”

A bibliometric mapping of advancements and trends in genome editing in pigs

“…Nevertheless, an impact assessment studies on future farming of IGAs are critical to devise strategies for its practical utility and adoption by end users and stakeholders. However, in spite of signi cant advancements in this eld, there are technical challenges like off-target effects, which have resulted in reduced viability and deformities in KO pigs (Chen et al, 2022). There exist regulatory, ethical and societal issues in channelizing the gene edited pigs from the boundaries of labs to the farmers eld and then to the food plate (Meyer and Vergnaud, 2021; Martin-Collado et al, 2022), nevertheless the consumer acceptance for gene edited food will be a major factor in deciding the fate of gene edited pigs (Martin-Collado et al, 2022).…”