2005 Fiscal Year Final Research Report Summary
Gene manipulation of hypoxia inducible gene regulation
| Project/Area Number |
16590923
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Hematology
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| Research Institution | University of Tsukuba |
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Principal Investigator |
IMAGAWA Shigehiko University of Tsukuba, GSCHS, Professor, 大学院・人間総合科学研究科, 教授 (60231164)
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| Co-Investigator(Kenkyū-buntansha) |
YAMAMOTO Masayuki University of Tsukuba, GSCHS, Professor, 大学院・人間総合科学研究科, 教授 (50166823)
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| Project Period (FY) |
2004 – 2005
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| Keywords | erythropoietin / HIF / GATA / VEGF / transcription factor / gene expression / promoter / enhancer |
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| Research Abstract |
Erythropoietin (Epo) gene expression is negatively regulated by GATA, which binds to the GATA site in the Epo promoter, and also positively regulated by hypoxia inducible factor-1 (HIF-1), which binds to the hypoxia responsive element (HRE) in the Epo enhancer. The disorders associated with anemia of chronic disease (ACD) are characterized by a deficiency of Epo, leading to Epo being considered as a treatment. Interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α), which increase the binding activity of GATA and inhibit Epo promoter activity, are increased in patients with ACD. In a previous study, we demonstrated the ability of K-7174 (a GATA-specific inhibitor) to improve Epo production that had been inhibited by IL-1β or TNF-α treatment in Hep3B cells in vitro and in an in vivo mouse assay (FASEB J 17:1742; 2003). However, K-7174 was injected intraperitoneally into the mice. We examined the ability of K-11706, a novel GATA-specific inhibitor that has a 1,000-fold higher affinity
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than K-7174, to improve Epo production following inhibition by IL-1β or TNF-α in Hep3B cells in vitro. Oral administration of K-11706 reversed the decreases of hemoglobin, serum Epo, reticulocyte counts and CFU-E induced by IL-1β or TNF-α. These results raise the possibility of using orally administered K-11706 as a novel drug for treating ACD (Blood 104:4300; 2004).
In oxygenated cells, hypoxia inducible factor-1 (HIF-1) α subunits are rapidly degraded by a mechanism that involves ubiquitination by the von Hippel-Lindau tumor suppressor E3 ligase complex using 2-oxoglutarate as a substrate. We examined the effect of 2-oxoglutarate on the production of Epo and vascular endothelial growth factor (VEGF). The expression of Epo and VEGF protein were dose-dependently downregulated in Hep3B cells by the addition of 2-oxoglutarate. The promoter activity of VEGF-luciferase was dose-dependently downregulated by the addition of 2-oxoglutarate. Gel mobility shift assays revealed that the addition of 2-oxoglutarate dose-dependently inhibited HIF-1 binding activity, but did not affect GATA binding activity. Western blot analysis revealed that 2-oxoglutarate dose-dependently inhibited the HIF-1α protein level in Hep3B cells in hypoxic conditions. However, MG132 (the proteasome inhibitor) rescued the inhibition of HIF-1α protein expression by 2-oxoglutarate. Furthermore, under hypoxic conditions, 2-oxoglutarate dose-dependently inhibited tube formation in in vitro angiogenesis assays. These results indicate that 2-oxoglutarate treatment may be useful for the inhibition of angiogenesis. Less
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[Journal Article] Enhanced erythropoiesis mediated by activation of the renin-angiotensin system via angiotensin II type la receptor.2005
Author(s)
Kato H, Ishida J, Imagawa S, Saito T, Suzuki N, Matsuoka T, Sugaya T, Tanimoto K, Yokoo T, Ohneda O, Sugiyama F, Yagami K, Fujita T, Yamamoto M, Nangaku M, Fukamizu A.
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Journal Title
FASEB J 19
Pages: 2023-2025
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Oral administration of K-11706 inhibits GATA binding activity, enhances hypoxia-inducible factor 1 binding activity and restores indicators in an in vivo mouse model of anemia of chronic disease.2004
Author(s)
Nakano Y, Imagawa S, Matsumoto K, Stockmann C, Obara N, Suzuki N, Doi T, Kodama T, Takahashi S, Nagasawa T, Yamamoto M.
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Journal Title
Blood 104
Pages: 4300-4307
Description
「研究成果報告書概要(欧文)」より
