title: Lampe1: an ENU-germline mutation causing spontaneous hepatosteatosis identified through targeted exon-enrichment and next-generation sequencing.
abstract: Using a small scale ENU mutagenesis approach we identified a recessive germline mutant, designated Lampe1 that exhibited growth retardation and spontaneous hepatosteatosis. Low resolution mapping based on 20 intercrossed Lampe1 mice revealed linkage to a ∼14 Mb interval on the distal site of chromosome 11 containing a total of 285 genes. Exons and 50 bp flanking sequences within the critical region were enriched with sequence capture microarrays and subsequently analyzed by next-generation sequencing. Using this approach 98.1 percent of the targeted DNA was covered with a depth of 10 or more reads per nucleotide and 3 homozygote mutations were identified. Two mutations represented intronic nucleotide changes whereas one mutation affected a splice donor site in intron 11-12 of Palmitoyl Acetyl-coenzyme A oxygenase-1 (Acox1), causing skipping of exon 12. Phenotyping of Acox1(Lampe1) mutants revealed a progression from hepatosteatosis to steatohepatitis, and ultimately hepatocellular carcinoma. The current approach provides a highly efficient and affordable method to identify causative mutations induced by ENU mutagenesis and animal models relevant to human pathology.
authors.affiliation: Department of Pathology and Laboratory Medicine, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States of America.
authors.forename: Rachel
authors.initials: R
authors.lastname: Sheridan
authors.forename: Kristin
authors.initials: K
authors.lastname: Lampe
authors.forename: Shiva Kumar
authors.initials: SK
authors.lastname: Shanmukhappa
authors.forename: Patrick
authors.initials: P
authors.lastname: Putnam
authors.forename: Mehdi
authors.initials: M
authors.lastname: Keddache
authors.forename: Senad
authors.initials: S
authors.lastname: Divanovic
authors.forename: Jorge
authors.initials: J
authors.lastname: Bezerra
authors.forename: Kasper
authors.initials: K
authors.lastname: Hoebe
mesh_terms: D000818:Animals; D005038:Ethylnitrosourea / Q000378:metabolism*; D005091:Exons / Q000235:genetics*; D005234:Fatty Liver / Q000150:complications / Q000235:genetics* / Q000473:pathology; D018390:Gene Targeting; D018095:Germ-Line Mutation / Q000235:genetics*; D006801:Humans; D050356:Lipid Metabolism; D008099:Liver / Q000002:abnormalities / Q000254:growth & development / Q000473:pathology / Q000648:ultrastructure; D008113:Liver Neoplasms / Q000150:complications / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D010641:Phenotype; D022821:RNA Splice Sites / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods*; D017384:Sequence Deletion / Q000235:genetics
chemical_list: D022821:RNA Splice Sites; D005038:Ethylnitrosourea