title: Mouse model for noninvasive imaging of HIF prolyl hydroxylase activity: assessment of an oral agent that stimulates erythropoietin production.
abstract: Many human diseases are characterized by the development of tissue hypoxia. Inadequate oxygenation can cause cellular dysfunction and death. Tissues use many strategies, including induction of angiogenesis and alterations in metabolism, to survive under hypoxic conditions. The heterodimeric transcription factor hypoxia-inducible factor (HIF) is a master regulator of genes that promote adaptation to hypoxia. HIF activity is linked to oxygen availability because members of the EGLN family hydroxylate HIFalpha subunits on specific prolyl residues when oxygen is present, which marks them for ubiquitination and proteasomal degradation. We created a mouse that ubiquitously expresses a bioluminescent reporter consisting of firefly luciferase fused to a region of HIF that is sufficient for oxygen-dependent degradation. Our validation studies suggest that this mouse will be useful for monitoring hypoxic tissues and evaluating therapeutic agents that stabilize HIF. One such agent, the HIF prolyl hydroxylase inhibitor FG-4383, was active in the liver and kidney after systemic administration as determined by bioluminescence imaging, transcription profiling, and production of erythropoietin, indicating that the HIF transcriptional program can be manipulated in vivo with orally active organic small molecules.
authors.affiliation: Department of Medical Oncology, Dana-Farber Cancer Institute, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Michal
authors.initials: M
authors.lastname: Safran
authors.forename: William Y
authors.initials: WY
authors.lastname: Kim
authors.forename: Fionnuala
authors.initials: F
authors.lastname: O'Connell
authors.forename: Lee
authors.initials: L
authors.lastname: Flippin
authors.forename: Volkmar
authors.initials: V
authors.lastname: Günzler
authors.forename: James W
authors.initials: JW
authors.lastname: Horner
authors.forename: Ronald A
authors.initials: RA
authors.lastname: Depinho
authors.forename: William G
authors.initials: WG
authors.lastname: Kaelin
mesh_terms: D000818:Animals; D015687:Cell Hypoxia / Q000502:physiology; D002478:Cells, Cultured; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D049308:Dioxygenases / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D004791:Enzyme Inhibitors; D004797:Enzyme-Linked Immunosorbent Assay; D004921:Erythropoietin / Q000096:biosynthesis*; D051793:Hypoxia-Inducible Factor 1 / Q000235:genetics / Q000378:metabolism*; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases; D008156:Luciferases; D051379:Mice; D023421:Models, Animal; D015336:Molecular Probe Techniques; D010957:Plasmids / Q000235:genetics; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D022661:RNA, Untranslated
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D004791:Enzyme Inhibitors; C110853:Gt(ROSA)26Sor non-coding RNA, mouse; D051793:Hypoxia-Inducible Factor 1; D011506:Proteins; D022661:RNA, Untranslated; D004921:Erythropoietin; D049308:Dioxygenases; D008156:Luciferases; C488894:EGLN3 protein, human; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases