title: Characterization of woodchuck apolipoprotein A-I: a new tool for drug delivery and identification of altered isoforms in the woodchuck chronic hepatitis model.
abstract: Apolipoprotein A-I (ApoA-I) is the major protein component of high density lipoprotein (HDL) particles in serum, and participates in the reverse transport of cholesterol from tissues to the liver for excretion. The natural HDL tropism to the liver and cancer cells has been used extensively to target encapsulated drugs. The alteration of the plasmatic isoforms of ApoA-I is a hallmark of chronic hepatitis and hepatocarcinoma in mice and humans. Woodchucks infected with the woodchuck hepatitis virus (WHV) represent the best animal model for the study of chronic viral hepatitis B and viral induced hepatocarcinoma (HCC). WHV-infected woodchuck represents a clinically relevant animal model under which new treatment strategies can be evaluated and optimized. Therapeutic efficacy in this model is likely to be translated into a successful therapy for patients infected with HBV. The present study describes, for the first time, the cloning and characterization of woodchuck ApoA-I. The open reading frame (ORF) of the woodchuck ApoA-I is 795 bp long, coding for 264 amino acids. Unexpectedly, phylogenetic analysis revealed that the closest sequences are those of human and macaque. Woodchuck HDLs were isolated successfully from sera by density gradient ultracentrifugation. A commercial antibody that recognized the woodchuck ApoA-I was also identified. Finally, taking advantage of the techniques and tools developed in this study, two potential applications of woodchuck HDLs are illustrated: drug delivery to a woodchuck hepatocarcinoma cell line and the use of isoelectrofocusing to identify ApoA-I isoforms.
authors.affiliation: Division of Hepatology and Gene Therapy, Center for Applied Medical Research, University of Navarra, Pamplona, Navarra, Spain.
authors.forename: Jessica
authors.initials: J
authors.lastname: Fioravanti
authors.forename: Celia
authors.initials: C
authors.lastname: Gomar
authors.forename: José
authors.initials: J
authors.lastname: Medina-Echeverz
authors.forename: Itziar
authors.initials: I
authors.lastname: Otano
authors.forename: Alberto
authors.initials: A
authors.lastname: Benito
authors.forename: Jesús
authors.initials: J
authors.lastname: Prieto
authors.forename: Gloria
authors.initials: G
authors.lastname: González-Aseguinolaza
authors.forename: Pedro
authors.initials: P
authors.lastname: Berraondo
mesh_terms: D000818:Animals; D016632:Apolipoprotein A-I / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D001483:Base Sequence; D002232:Carbocyanines / Q000032:analysis; D006528:Carcinoma, Hepatocellular; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D004195:Disease Models, Animal; D016503:Drug Delivery Systems / Q000379:methods; D005434:Flow Cytometry; D017919:Hepatitis B Virus, Woodchuck / Q000254:growth & development; D019694:Hepatitis B, Chronic / Q000235:genetics* / Q000378:metabolism / Q000473:pathology / Q000821:virology*; D006801:Humans; D008099:Liver / Q000473:pathology / Q000821:virology; D008113:Liver Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology / Q000821:virology*; D008392:Marmota / Q000235:genetics / Q000378:metabolism / Q000821:virology*; D051379:Mice; D008969:Molecular Sequence Data; D058990:Molecular Targeted Therapy / Q000379:methods; D020033:Protein Isoforms / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014162:Transfection; D014779:Virus Replication
chemical_list: C035005:3,3'-dihexadecylindocarbocyanine; D016632:Apolipoprotein A-I; D002232:Carbocyanines; D020033:Protein Isoforms