Conditional inactivation of HIF-1 using intrabodies.
Hypoxia is a hallmark of solid cancers and triggers the transcription of genes responsible for cell survival. The transcription factor Hypoxia-Inducible Factor 1 (HIF-1) is a key regulator in this response and frequently activated in human cancer. HIF-1 activation is associated with tumor aggressiveness and poor clinical outcome and, therefore, may provide an attractive therapeutic target. Here we provide a novel approach for HIF-1 targeted therapy using single-domain llama antibodies directed against the HIF-1alpha oxygen dependent degradation domain which encompass the N-terminal transactivation domain. Conditional expression of HIF intrabodies in mammalian cells interfered with binding to pVHL and inhibited hypoxia induced activation of endogenous target genes. Inducible intrabody targeting is a highly specific strategy for temporal protein inactivation and may have applications for disease treatment.
Groot
Arjan J
AJ
Department of Pathology, University Medical Center Utrecht, 3508 GA, Utrecht, The Netherlands.
Gort
Eelke H
EH
van der Wall
Elsken
E
van Diest
Paul J
PJ
Vooijs
Marc
M
D000818:Animals; D000915:Antibody Affinity; D000918:Antibody Specificity; D015153:Blotting, Western; D002161:Camelids, New World; D045744:Cell Line, Tumor; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D007128:Immunoglobulin Fragments / Q000276:immunology*; D047468:Immunoprecipitation; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic; D014162:Transfection
D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D007128:Immunoglobulin Fragments