Basic–Alimentary TractMucosal Protection by Hypoxia-Inducible Factor Prolyl Hydroxylase Inhibition
Section snippets
Cell Culture
HeLa cells were grown and maintained as confluent monolayers on 24-well plates (Costar Corp, Cambridge, MA). Cell cultures were exposed to hypoxia as described previously.15
FG-4442 and FG-4497 were synthesized at FibroGen, Inc (San Francisco, CA). These small molecule inhibitors of PHD enzymes were previously disclosed in patent filings US006093730A and US20040254215A1, respectively. Where indicated, cells were exposed to the PHD inhibitors FG-4497 or FG-4442 at indicated concentrations for
FG-4442 and FG-4497 Increase HIF-1 Activity In Vitro and In Vivo
Initially, we sought to characterize the ability of 2 new HIF PHD inhibitors (FG-4442 and FG-4497) to activate HIF in vitro and to examine expression of HIF target genes in vivo. As shown in Figure 1, using a HIF reporter plasmid transiently transfected into HeLa cells, we showed that FG-4442 increased HIF activation in a concentration-dependent fashion (P < .025), with maximal HIF activity increases of 5.1- ± 0.8-fold (P < .01). In this regard, FG-4497 was more potent that FG-4442. As shown in
Discussion
These studies provide new insight into the potential therapeutic use of PHD inhibitors on the acute phases of mucosal inflammation. On the basis of previous work implicating a protective role for HIF-1α in mucosal inflammatory disease, we focused on defining epithelial HIF activation and colitis end points. These studies extend previous work into the role of HIF in vivo and provide new insight into the potential use of PHD inhibitors as therapeutic modalities.
HIF protein expression and activity
References (45)
- et al.
Hypoxia-inducible factor 1alpha (HIF-1alpha) protein is rapidly degraded by the ubiquitin-proteasome system under normoxic conditionsIts stabilization by hypoxia depends on redox-induced changes
J Biol Chem
(1997) - et al.
Molecular mechanism of hypoxia-inducible factor 1alpha -p300 interactionA leucine-rich interface regulated by a single cysteine
J Biol Chem
(2001) - et al.
Hapten-induced model of chronic inflammation and ulceration in the rat colon
Gastroenterology
(1989) - et al.
Matrix metalloproteinase activity synergizes with alpha2beta1 integrins to enhance collagen remodeling
Exp Cell Res
(2005) HIF-1, O(2), and the 3 PHDs: how animal cells signal hypoxia to the nucleus
Cell
(2001)Recent advances in erythropoietic agents in renal anemia
Semin Nephrol
(2006)- et al.
Critical role of cAMP response element binding protein expression in hypoxia-elicited induction of epithelial tumor necrosis factor-alpha
J Biol Chem
(1999) - et al.
Ecto-nucleotidases--molecular structures, catalytic properties, and functional roles in the nervous system
Prog Brain Res
(1999) - et al.
Regulation of angiogenesis by hypoxia and hypoxia-inducible factors
Curr Top Dev Biol
(2006) - et al.
Hypoxia, HIF-1, and the pathophysiology of common human diseases
Adv Exp Med Biol
(2000)
Epithelial hypoxia-inducible factor-1 is protective in murine experimental colitis
J Clin Invest
Hypoxia-inducible factor 1-dependent induction of intestinal trefoil factor protects barrier function during hypoxia
J Exp Med
Crucial role for ecto-5’-nucleotidase (CD73) in vascular leakage during hypoxia
J Exp Med
Hypoxia-inducible factor-1-dependent regulation of the multidrug resistance (MDR1) gene
Cancer Res
Cellular adaptation to hypoxia: O2-sensing protein hydroxylases, hypoxia-inducible transcription factors, and O2-regulated gene expression
FASEB J
HIFalpha targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing
Science
Targeting of HIF-alpha to the von Hippel-Lindau ubiquitylation complex by O2-regulated prolyl hydroxylation
Science
HIF-1alpha binding to VHL is regulated by stimulus-sensitive proline hydroxylation
Proc Natl Acad Sci U S A
Regulation of hypoxia-inducible factor 1alpha is mediated by an O2-dependent degradation domain via the ubiquitin-proteasome pathway
Proc Natl Acad Sci U S A
Hypoxia-inducible factor-1 modulates gene expression in solid tumors and influences both angiogenesis and tumor growth
Proc Natl Acad Sci U S A
Molecular mechanisms of transcription activation by HLF and HIF1alpha in response to hypoxia: their stabilization and redox signal-induced interaction with CBP/p300
EMBO J
Epithelial exposure to hypoxia modulates neutrophil transepithelial migration
J Exp Med
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All authors declare that they have no conflict of interest to disclose.
Supported by National Institutes of Health grants HL60569, DE016191, and DK50189 and by a grant from the Crohn’s and Colitis Foundation of America.