Investigation of Genetic Modifiers of Copper Toxicosis in Labrador Retrievers

Abstract

Copper toxicosis is a complex genetic disorder in Labrador retrievers characterized by hepatic copper accumulation eventually leading to liver cirrhosis. The variation of hepatic copper levels in Labrador retrievers has been partly explained by mutations in ATP7A c.980C>T and ATP7B c.4358G>A. To further elucidate the genetic background of this disease, we used targeted Next Generation Sequencing (NGS) in a cohort of 95 Labrador retrievers to analyze 72 potential modifier genes for variations associated with hepatic copper levels. Variants associated with copper levels were subsequently evaluated in a replication cohort of 144 Labrador retrievers. A total of 44 variants in 25 different genes were identified, of which four showed significant association with copper levels. Of the four variants found associated with hepatic copper levels in the NGS cohort, one was validated in the replication cohort. The non-reference allele of the variant NC_006602.3.g.52434480C>T in RETN resulting in amino-acid change p.Leu7Phe was associated with decreased hepatic copper levels. In humans, resistin is associated with severity of non-alcoholic fatty liver disease, fibrosis, cirrhosis and mitochondrial dysfunction in hepatocytes. Further studies are needed to investigate the biological function of RETN p.Leu7Phe in the development of copper toxicosis in Labrador retrievers.

Keywords: ATP7A; ATP7B; Labrador retriever; Menkes disease; RETN; Wilson disease; copper; copper toxicosis; dog.

Figure 1

Boxplots of copper score associations between different genotypes in the NGS and replication cohort. (A,B) Boxplots show a positive significant association between copper score and the non-reference A allele for the variation rs85069010 in NPC1 in the NGS but not in the replication cohort. (C,D) Boxplots show a significant negative association between copper score and the non-reference T allele for the variation rs852470997 in RETN in both the NGS and replication cohort. (E,F) Boxplots show a significant negative association between copper score and the non-reference C allele for the variation NC_002008.4:MT.7593T>T in MT-CO2 in the NGS but not in the replication cohort.