title: Directed evolution of gene-shuffled IFN-alpha molecules with activity profiles tailored for treatment of chronic viral diseases.
abstract: Type I IFNs are unusually pleiotropic cytokines that bind to a single heterodimeric receptor and have potent antiviral, antiproliferative, and immune modulatory activities. The diverse effects of the type I IFNs are of differential therapeutic importance; in cancer therapy, an enhanced antiproliferative effect may be beneficial, whereas in the therapy of viral infections (such as hepatitis B and hepatitis C), the antiproliferative effects lead to dose limiting bone marrow suppression. Studies have shown that various members of the natural IFN-alpha family and engineered variants, such as IFN-con1, vary in the ratios between various IFN-mediated cellular activities. We used DNA shuffling to explore and confirm the hypothesis that one could simultaneously increase the antiviral and Th1-inducing activity and decrease the antiproliferative activity. We report IFN-alpha hybrids wherein the ratio of antiviral:antiproliferative and Th1-inducing: antiproliferative potencies are markedly increased with respsect to IFN-con1 (75- and 80-fold, respectively). A four-residue motif that overlaps with the IFNAR1 binding site and is derived by cross breeding with a pseudogene contributes significantly to this phenotype. These IFN-alphas have an activity profile that may result in an improved therapeutic index and, consequently, better clinical efficacy for the treatment of chronic viral diseases such as hepatitis B virus, human papilloma virus, HIV, or chronic hepatitis C.
authors.affiliation: Maxygen, Incorporated, 515 Galveston Drive, Redwood City, CA 94063, USA.
authors.forename: Amy D
authors.initials: AD
authors.lastname: Brideau-Andersen
authors.forename: Xiaojian
authors.initials: X
authors.lastname: Huang
authors.forename: Siu-Chi Chang
authors.initials: SC
authors.lastname: Sun
authors.forename: Teddy T
authors.initials: TT
authors.lastname: Chen
authors.forename: Diane
authors.initials: D
authors.lastname: Stark
authors.forename: Ian J
authors.initials: IJ
authors.lastname: Sas
authors.forename: Linda
authors.initials: L
authors.lastname: Zadik
authors.forename: Glenn N
authors.initials: GN
authors.lastname: Dawes
authors.forename: Douglas R
authors.initials: DR
authors.lastname: Guptill
authors.forename: Robert
authors.initials: R
authors.lastname: McCord
authors.forename: Sridhar
authors.initials: S
authors.lastname: Govindarajan
authors.forename: Ajoy
authors.initials: A
authors.lastname: Roy
authors.forename: Shumin
authors.initials: S
authors.lastname: Yang
authors.forename: Judy
authors.initials: J
authors.lastname: Gao
authors.forename: Yong Hong
authors.initials: YH
authors.lastname: Chen
authors.forename: Niels Jørgen Ø
authors.initials: NJ
authors.lastname: Skartved
authors.forename: Annette K
authors.initials: AK
authors.lastname: Pedersen
authors.forename: David
authors.initials: D
authors.lastname: Lin
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Locher
authors.forename: Indrani
authors.initials: I
authors.lastname: Rebbapragada
authors.forename: Anne Dam
authors.initials: AD
authors.lastname: Jensen
authors.forename: Steven H
authors.initials: SH
authors.lastname: Bass
authors.forename: Torben L Straight
authors.initials: TL
authors.lastname: Nissen
authors.forename: Sridhar
authors.initials: S
authors.lastname: Viswanathan
authors.forename: Graham R
authors.initials: GR
authors.lastname: Foster
authors.forename: Julian A
authors.initials: JA
authors.lastname: Symons
authors.forename: Phillip A
authors.initials: PA
authors.lastname: Patten
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D016466:CHO Cells; D002908:Chronic Disease / Q000628:therapy*; D006224:Cricetinae; D003412:Cricetulus; D032621:DNA Shuffling; D019020:Directed Molecular Evolution; D015723:Gene Library; D006367:HeLa Cells; D006801:Humans; D016898:Interferon-alpha / Q000737:chemistry / Q000235:genetics* / Q000494:pharmacology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D011544:Pseudogenes; D018417:Th1 Cells / Q000187:drug effects; D014777:Virus Diseases / Q000628:therapy*
chemical_list: D000998:Antiviral Agents; D016898:Interferon-alpha