Directed evolution of gene-shuffled IFN-alpha molecules with activity profiles tailored for treatment of chronic viral diseases.
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.
Brideau-Andersen
Amy D
AD
Maxygen, Incorporated, 515 Galveston Drive, Redwood City, CA 94063, USA.
Huang
Xiaojian
X
Sun
Siu-Chi Chang
SC
Chen
Teddy T
TT
Stark
Diane
D
Sas
Ian J
IJ
Zadik
Linda
L
Dawes
Glenn N
GN
Guptill
Douglas R
DR
McCord
Robert
R
Govindarajan
Sridhar
S
Roy
Ajoy
A
Yang
Shumin
S
Gao
Judy
J
Chen
Yong Hong
YH
Skartved
Niels Jørgen Ø
NJ
Pedersen
Annette K
AK
Lin
David
D
Locher
Christopher P
CP
Rebbapragada
Indrani
I
Jensen
Anne Dam
AD
Bass
Steven H
SH
Nissen
Torben L Straight
TL
Viswanathan
Sridhar
S
Foster
Graham R
GR
Symons
Julian A
JA
Patten
Phillip A
PA
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*
D000998:Antiviral Agents; D016898:Interferon-alpha