1999 Fiscal Year Annual Research Report

炎症性増殖性血管病変の分子病態解明と遺伝子治療法の開発-組換えアデノウイルスを用いた血管壁・心臓への生体内遺伝子導入による病態解析と治療効果の検討-

Research Project

Project/Area Number	09470170
Research Institution	KYUSHU UNIVERSITY
Principal Investigator	上野光九州大学, 医学部, 講師 (50260378)
Co-Investigator(Kenkyū-buntansha)	西田誉浩九州大学, 医学部, 助手 (50284500) 大原郁一九州大学, 医学部, 助手 (90185364) 筒井裕之九州大学, 医学部, 助手 (70264017) 居石克夫九州大学, 大学院・医学系研究科, 教授 (70108710) 盛栄三東海大学, 医学部, 助教授 (90146598)
Keywords	遺伝子導入 / アデノウイルス / 変異型TGFーβ受容体 / TGF-β / 細胞外マトリクス / TFPI / 線維化
Research Abstract	1.Tissue factor pathway inhibitor(TFPI)産生アデノウイルスを作製し、バルーンで障害を加えたウサギ頚動脈壁に導入した。導入7日後に、血管狭窄によりシアストレスを加えても血栓形成は惹起されなかった。この血栓形成抑制効果はカテコラミン存在下でも全く影響を受けなかった。全身の凝固・線溶系は影響を受けず血小板機構も正常に保持された。TFPIの局所遺伝子導入により出血性副作用を伴わない理想的な抗血栓遺伝子療法への展開が期待できる。成果は米国心臓病学会誌に発表したが、"Blue print for future local gene therapy"とのEditorial commentを得た。 2.CNP遺伝子を針付きカテーテルを用いてブタ冠動脈に導入したところ内膜肥厚の抑制が達成された。 3.細胞外マトリックスの産生・蓄積が血管壁の硬化と内腔の狭窄の主因であり、この過程はTGF-βで制御されている可能性が高い。TGF-βの作用を特異的に抑制できる変異型TGF-β受容体産生ベクタを作製し、肝臓線維症モデルに導入した。すると肝線維化が著明に抑制され、血清中の肝細胞酵素は低値に保たれ、肝不全から全例救命できた。線維性病変形成におけるTGF-βの中心的役割を実証するとともに線維化の抑制が治療的であることを初めて明らかに示した。傷害血管壁への導入実験を開始した。 4.受容体の細胞外ドメインのみ可溶型受容体を過剰発現させるとリガンドを吸着してその作用を抑制できる可能性がある。肝臓線維化モデルで検証したところ、変異型受容体と同様の抑制効果を観察し得た。この戦略では標的臓器の一部の細胞に遺伝子導入されればよいばかりか、全く離れた部位への導入でもよいことになり、遺伝子治療を想定する際には重要な知見である。すべての血管壁細胞に遺伝子導入されなくてもよいことを意味している。

Research Products
(28 results)

All Other

All Publications (28 results)

[Publications] Honda M, et al: "Experimental subretinal neovascularization is inhibited by adenovirus-mediated soluble VEGF/flt-1 receptor gene transfection:A role of VEGF possible treatment for SRN in age-related macular degeneration."Gene Ther. (in press). (2000)
[Publications] Sakamoto T, et al.: "Blockade of TGF-β by in vivo gene transfer of a soluble TGF-βtype II receptor in the muscle inhibits corneal opacification, edema and angiogenesis."Lab. Invest. (in press). (2000)
[Publications] Tremain R, et al.: "Defectcts in TGF-β signaling overcome senescence of mouse keratinocytes expressing v-H-ras."Oncogene. (in press). (2000)
[Publications] Uruno T, et al.: "Distinct regulation of myoblast defferentiation by intracellular and extracellular fibroblrast growth factor-1."Growth Factors. 17. 93-113 (1999)
[Publications] Glick A, et al.: "Defects in TGF-βsignaling cooperate with a Ras oncogene to cause rapid aneuploidy and malignant transformation of mouse keratinocytes. Proc."Natl. Acad. Sci. 96. 14949-14954 (1999)
[Publications] Eguchi S. et al.: "Intracellular signaling of angiotensin II-induced p70 S6 kinase phosphorylation at Ser 411 in vascular smooth muscle cells."J. Biol. Chem.. 274. 36843-36851 (1999)
[Publications] Saadeh PB, et al.: "Transforming growth factor beta 1 modulates the expression of VEGF by osteoblasts."Am. J. Physiol.. (in press). (1999)
[Publications] Steinbrech DS, et al.: "Mechanisms of FGF-2 modulation of VEGF expression by osteoblasts."Endocrinology. (in press). (1999)
[Publications] Takayama K, et al.: "Suppression of tumor angiogeesis and growth by gene transfer of a soluble form of VEGF receptor into a remote organ."Cancer Res.. (in press). (2000)
[Publications] Ueno H, et al.: "A soluble TGF-β receptor expressed in muscle prevents liver fibrogenesis and dysfunction in rats."Hum Gene Ther.. 11. 33-42 (2000)
[Publications] Morishige K, et al.: "Takeshita A. Local adnovirus-mediated transfer of C-type natriuetic peptide suppresses vascular remodeling in porcine coronary arteries in vivo."J. Am. Coll. Cardiol.. (in press). (2000)
[Publications] Kotani K, et al.: "Dominant-negative forms of Akt(protein kinase B) and atypicak PKCδ do not prevent insulin inhibition of PEPCK gene transcription."J. Biol. Chem.. 274. 21305-21312 (1999)
[Publications] Iwasaki H, et al.: "Endothelin-mediated bascular growth requires p42/p44 MAP kinase and p70 S6 kinase cascades via transactivation of epidermal growth factor receptor."Endocrinology. 140. 4659-4668 (1999)
[Publications] Saiki M, et al.: "Adenovirus-mediated gene transfer of a truncated form of fibroblast growth factor receptor inhibits growth of glioma cells both in vitro and in vivo."J. Neuro-Oncology. (in press). (1999)
[Publications] Nishida T, et al.: "Adenovirus-mediated local expression of human tissue factor pathway inhibitor eliminates shear stress-induced recurrent thrombosis in the injured carotid artery of the rabbit."Circ Res. 84. 1446-1452 (1999)
[Publications] Matsumoto T, et al.: "Plantelet-derived growth factor activates p38 mitogen activated protein kinase through ras- dependent pathway which is important for actin reorganization and cell migration."J. Biol Chem. 274. 13954-13960 (1999)
[Publications] Yamamoto H, et al.: "Roles of H-ras in signaling by TGF-β:H-ras is essential in activation of MAP kinase, partially involved in transcriptional activation by TGF-β but not in signaling of anti-proliferative response of TGF-β."Eur. J. Biochem. 264. 110-119 (1999)
[Publications] Takahashi T, et al.: "VEGF activates protein kinase C-dependent, but Ras-independent Raf-MEK-MAP kinase pathway fro DNA synthesis in primary endothelial cells."Oncogene. 18. 2221-2230 (1999)
[Publications] Qi Z, et al.: "Blockad of TGF-β signaling prevents liver fibrosis and dysfunction in the rat."Proc. Natl. Acad. Sci. USA. 96. 2221-2230 (1999)
[Publications] Yukawa H, et al.: "In vitro growth suppression of vascular smooth muscle cells using adenovirus-mediated gene transfer of a truncated form of fibroblast growth factor receptor."Atherosclerosis. 141. 125-132 (1998)
[Publications] Li Q et al.: "Stretch-induced collagen synthesis in culturede smooth muscle cells from rabbit aortic media and a possible involvement of angiotensis II and transforming growth factor-β."Vas. Res. 35. 93-103 (1998)
[Publications] Maru Y, et al.: "Shibuya M. Virally activated Ras cooperates with integrin to induce tubulogenesis in sinusoidal endothelial cell lines"J. Cell. Physiol. 176. 223-234 (1998)
[Publications] Li Q et al.: "Stretch-induced proliferation of cultured vascular smooth muscle cells and a possible involvement of local renin-angiotensis system and platelet-derived grwoth factor."Hypertens. Res. 20. 217-223 (1997)
[Publications] Ueno H, et al.: "Adeovirus-mediated expression of secreted form of basic fibroblast growth factor(FGF-2) induces cellular proliferation and angiogenesis in vivo."Arterioscl. Thromb. Vasc, Biol.. 17. 2453-2460 (1997)
[Publications] Ueno H, et al.: "Local expressison of C-type natriuretic peptide markedly suppresses neointimal formation in rat injured arteries through an autocrine/paracrine loop."Circulation. 96. 2272-2279 (1997)
[Publications] Ueno H, et al.: "Adenovirus-mediated transfer of a dominant-negative H-ras suppresses neointimal formation in balloon-injured arteries in vivo."Arterioscl. Thromb. Vasc, Biol.. 17. 898-904 (1997)
[Publications] Takahashi T, et al.: "Angiotensin II stimulates mitogen-activated protein syntesis by a Ras-independent pathway in vascular smooth muscle sells."J. Biol. Chem.. 272. 16018-16022 (1997)
[Publications] Ueno H, et al.: "Adenovirus-mediated transfer of cyclin-dependent kinase inhibitor-p21 suppresses neointimal formation in the balloon-injured rat carotid arteries in vivo."Ann. NY. Acad. Sci.. 811. 40-411 (1997)

1999 Fiscal Year Annual Research Report

炎症性増殖性血管病変の分子病態解明と遺伝子治療法の開発-組換えアデノウイルスを用いた血管壁・心臓への生体内遺伝子導入による病態解析と治療効果の検討-

Principal Investigator

上野 光 九州大学, 医学部, 講師 (50260378)

Research Products

[Publications] Honda M, et al: "Experimental subretinal neovascularization is inhibited by adenovirus-mediated soluble VEGF/flt-1 receptor gene transfection:A role of VEGF possible treatment for SRN in age-related macular degeneration."Gene Ther. (in press). (2000)

[Publications] Sakamoto T, et al.: "Blockade of TGF-β by in vivo gene transfer of a soluble TGF-βtype II receptor in the muscle inhibits corneal opacification, edema and angiogenesis."Lab. Invest. (in press). (2000)

[Publications] Tremain R, et al.: "Defectcts in TGF-β signaling overcome senescence of mouse keratinocytes expressing v-H-ras."Oncogene. (in press). (2000)

[Publications] Uruno T, et al.: "Distinct regulation of myoblast defferentiation by intracellular and extracellular fibroblrast growth factor-1."Growth Factors. 17. 93-113 (1999)

[Publications] Glick A, et al.: "Defects in TGF-βsignaling cooperate with a Ras oncogene to cause rapid aneuploidy and malignant transformation of mouse keratinocytes. Proc."Natl. Acad. Sci. 96. 14949-14954 (1999)

[Publications] Eguchi S. et al.: "Intracellular signaling of angiotensin II-induced p70 S6 kinase phosphorylation at Ser 411 in vascular smooth muscle cells."J. Biol. Chem.. 274. 36843-36851 (1999)

[Publications] Saadeh PB, et al.: "Transforming growth factor beta 1 modulates the expression of VEGF by osteoblasts."Am. J. Physiol.. (in press). (1999)

[Publications] Steinbrech DS, et al.: "Mechanisms of FGF-2 modulation of VEGF expression by osteoblasts."Endocrinology. (in press). (1999)

[Publications] Takayama K, et al.: "Suppression of tumor angiogeesis and growth by gene transfer of a soluble form of VEGF receptor into a remote organ."Cancer Res.. (in press). (2000)

[Publications] Ueno H, et al.: "A soluble TGF-β receptor expressed in muscle prevents liver fibrogenesis and dysfunction in rats."Hum Gene Ther.. 11. 33-42 (2000)

[Publications] Morishige K, et al.: "Takeshita A. Local adnovirus-mediated transfer of C-type natriuetic peptide suppresses vascular remodeling in porcine coronary arteries in vivo."J. Am. Coll. Cardiol.. (in press). (2000)

[Publications] Kotani K, et al.: "Dominant-negative forms of Akt(protein kinase B) and atypicak PKCδ do not prevent insulin inhibition of PEPCK gene transcription."J. Biol. Chem.. 274. 21305-21312 (1999)

[Publications] Iwasaki H, et al.: "Endothelin-mediated bascular growth requires p42/p44 MAP kinase and p70 S6 kinase cascades via transactivation of epidermal growth factor receptor."Endocrinology. 140. 4659-4668 (1999)

[Publications] Saiki M, et al.: "Adenovirus-mediated gene transfer of a truncated form of fibroblast growth factor receptor inhibits growth of glioma cells both in vitro and in vivo."J. Neuro-Oncology. (in press). (1999)

[Publications] Nishida T, et al.: "Adenovirus-mediated local expression of human tissue factor pathway inhibitor eliminates shear stress-induced recurrent thrombosis in the injured carotid artery of the rabbit."Circ Res. 84. 1446-1452 (1999)

[Publications] Matsumoto T, et al.: "Plantelet-derived growth factor activates p38 mitogen activated protein kinase through ras- dependent pathway which is important for actin reorganization and cell migration."J. Biol Chem. 274. 13954-13960 (1999)

[Publications] Yamamoto H, et al.: "Roles of H-ras in signaling by TGF-β:H-ras is essential in activation of MAP kinase, partially involved in transcriptional activation by TGF-β but not in signaling of anti-proliferative response of TGF-β."Eur. J. Biochem. 264. 110-119 (1999)

[Publications] Takahashi T, et al.: "VEGF activates protein kinase C-dependent, but Ras-independent Raf-MEK-MAP kinase pathway fro DNA synthesis in primary endothelial cells."Oncogene. 18. 2221-2230 (1999)

[Publications] Qi Z, et al.: "Blockad of TGF-β signaling prevents liver fibrosis and dysfunction in the rat."Proc. Natl. Acad. Sci. USA. 96. 2221-2230 (1999)

[Publications] Yukawa H, et al.: "In vitro growth suppression of vascular smooth muscle cells using adenovirus-mediated gene transfer of a truncated form of fibroblast growth factor receptor."Atherosclerosis. 141. 125-132 (1998)

[Publications] Li Q et al.: "Stretch-induced collagen synthesis in culturede smooth muscle cells from rabbit aortic media and a possible involvement of angiotensis II and transforming growth factor-β."Vas. Res. 35. 93-103 (1998)

[Publications] Maru Y, et al.: "Shibuya M. Virally activated Ras cooperates with integrin to induce tubulogenesis in sinusoidal endothelial cell lines"J. Cell. Physiol. 176. 223-234 (1998)

[Publications] Li Q et al.: "Stretch-induced proliferation of cultured vascular smooth muscle cells and a possible involvement of local renin-angiotensis system and platelet-derived grwoth factor."Hypertens. Res. 20. 217-223 (1997)

[Publications] Ueno H, et al.: "Adeovirus-mediated expression of secreted form of basic fibroblast growth factor(FGF-2) induces cellular proliferation and angiogenesis in vivo."Arterioscl. Thromb. Vasc, Biol.. 17. 2453-2460 (1997)

[Publications] Ueno H, et al.: "Local expressison of C-type natriuretic peptide markedly suppresses neointimal formation in rat injured arteries through an autocrine/paracrine loop."Circulation. 96. 2272-2279 (1997)

[Publications] Ueno H, et al.: "Adenovirus-mediated transfer of a dominant-negative H-ras suppresses neointimal formation in balloon-injured arteries in vivo."Arterioscl. Thromb. Vasc, Biol.. 17. 898-904 (1997)

[Publications] Takahashi T, et al.: "Angiotensin II stimulates mitogen-activated protein syntesis by a Ras-independent pathway in vascular smooth muscle sells."J. Biol. Chem.. 272. 16018-16022 (1997)

[Publications] Ueno H, et al.: "Adenovirus-mediated transfer of cyclin-dependent kinase inhibitor-p21 suppresses neointimal formation in the balloon-injured rat carotid arteries in vivo."Ann. NY. Acad. Sci.. 811. 40-411 (1997)

上野光九州大学, 医学部, 講師 (50260378)