Establishment of new therapeutic strategies for neurogenetic disorders during childhood

• Project/Area Number: 16390302
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Pediatrics
• Research Institution: Tottori University
• Principal Investigator: OHNO Kousaku Tottori University, Faculty of Medicine, Professor, 医学部, 教授 (70112109)
• Co-Investigator(Kenkyū-buntansha): NINOMIYA Haruaki Tottori University, Faculty of Medicine, Associate Professor, 医学部, 助教授 (80212124) ; INOUE Takehiko Tottori University, University Hospital, Research Associate, 医学部附属病院, 助手 (80346361)
• Project Period (FY): 2004 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥13,800,000 (Direct Cost: ¥13,800,000); Fiscal Year 2006: ¥4,600,000 (Direct Cost: ¥4,600,000); Fiscal Year 2005: ¥4,100,000 (Direct Cost: ¥4,100,000); Fiscal Year 2004: ¥5,100,000 (Direct Cost: ¥5,100,000)
• Keywords: Gaucher disease / Chemical chaperone therapy / N-octyl-β-valienamine / Niemann-Pick disease C / cholesterol / Toll-like receptor / IL-6 / glia / ニーマン・ピック病 / ニーマンピック病 / ライソゾーム病 / 酵素蛋白質の安定化 / βグルコシダーゼ

Research Abstract

Gaucher disease (GD), caused by a defect of β-glucosidase (β-Glu), is the most common form of sphingolipidosis. We found that a carbohydrate mimic β-octyl-β-valienamine (NOV), an inhibitor of β-Glu, could increase the protein level and enzyme activity of F213I, N188S, G202R and N370S mutant forms of β-Glu in cultured cells. When expressed in COS cells, the mutant proteins as well as the wild-type protein were localized predominantly in the endoplasmic reticulum and were sensitive to Endo-H treatment. NOV did not alter this localization or Endo-H sensitivity, suggesting that it acted in the endoplasmic reticulum. Profiling of β-alkyl-β-valienamines with various lengths of the acyl chain showed that β-dodecyl-β-valienamine was as effective as NOV. These results suggest a potential therapeutic value of NOV and related compounds for GD with a broad range of β-Glu mutations.
Niemann-Pick disease type C (NPC) is a lipid storage disorder caused by mutations in NPC1 (NPC2) genes. We found that human NPC fibroblasts secrete several cytokines and contain increased levels of STATs. These cells also contained increased levels of TLR4. In the NPC model mouse brain, glial cells expressed TLR4 and IL-6, whereas both glial and neuronal cells expressed STATs. Genetic deletion of TLR4 in NPC model mice reduced IL-6 secretion but failed to alter STAT levels or glial cell activation in the brain. In contrast, genetic deletion of IL-6 normalized STAT levels and suppressed glial cell activation. These findings indicate that constitutive cytokine secretion leads to activation of STATs and that this secretion is partly caused by an endosomal accumulation of TLR4. These results also suggest that similar signaling events may underlie glial cell activation in the NPC brain

Research Products

• [Journal Article] Endosomal accumulation of Toll-like receptor 4 causes constitutive secretion of cytokines and activation of STATs in Niemann-Pick disease type C fibroblasts : a potential basis for glial cell activation in the NPC brain. 27 : 1879-1891,2007 (2007)
  • Author(s): Suzuki M, Sugimoto Y, Ohsaki Y, Ueno M, Kato S, Kitamura Y, Hosokawa H, Davies JP, Ioannou YA, Vanier MT, Ohno K, Ninomiya H
  • Journal Title: Journal of Neuroscience 27 Pages: 1879-1891
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Enzyme enhancement activity of N-octyl-beta-valienamine on beta-glucosidase mutants associated with Gaucher disease. (2007)
  • Author(s): Lei K, Ninomiya H, Suzuki M, Inoue T, Sawa M, Iida M, Ida H, Eto Y, Ogawa S, Ohno K, Suzuki Y
  • Journal Title: Biochim Biophys Acta 1772 Pages: 587-596
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Enzyme enhancement activity of N-octyl-beta-valienamine on beta-glucosidase mutants associated with Gaucher disease. (2007)
  • Author(s): Lei K, Ninomiya H, Suzuki M, Inoue T, Sawa M, Iida M, Ida H, Eto Y, Ogawa S, Ohno K, Suzuki Y.
  • Journal Title: Biochim Biophys Acta. 1772 Pages: 587-96
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Endosomal accumulation of Toll-like receptor 4 causes constitutive secretion of cytokines and activation of STATs in Niemann-Pick disease type C fibroblasts : a potential basis for glial cell activation in the NPC brain. (2007)
  • Author(s): Suzuki M, Sugimoto Y, Ohsaki Y, Ueno M, Kato S, Kitamura Y, Hosokawa H, Davies JP, Ioannou YA, Vanier MT, Ohno K, Ninomiya H.
  • Journal Title: J Neurosci 27 Pages: 1879-1891
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Endosomal Accumulation of Toll-like receptor 4 causes constitutive secretion of cytokines and activation of STATs in Niemann-Pick disease type C fibroblasts : a potential basis for glial cell activation in the NPC brain. (2007)
  • Author(s): Suzuki M, et al.
  • Journal Title: J Neurosci 27 Pages: 1879-1891
• [Journal Article] Cholesterol depletion facilitates ubiquitylation of NPC1 and its association with SKD1/Vps4. (2006)
  • Author(s): Ohsaki Y, Sugimoto Y, Suzuki M, Hosokawa H, Yoshimori T, Davies JP, Ioannou YA, Vanier MT, Ohno K, Ninomiya H
  • Journal Title: J Cell Sci 119 Pages: 2643-2653
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Chemical synthesis of the 3-sulfooxy-7-N-acetylglucosaminyl-24- amidated conjugates of 3β,7β-dihydroxy-5-cholen-24-oic acid, and related compounds : Unusual, major metabolites of bile acid in a patient with Niemann-Pick disease type C1. (2006)
  • Author(s): Iida T, Kakiyama G, Hibiya Y, Miyata S, Inoue T, Ohno K, Goto T, Mano N, Goto J, Nambara T, A F Hofmann
  • Journal Title: Steroids 71 Pages: 18-29
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Cholesterol depletion facilitates ubiquitylation of NPC1 and its association with SKD1/Vps4. (2006)
  • Author(s): Ohsaki Y, Sugimoto Y, Suzuki M, Hosokawa H, Yoshimori T, Davies JP, Ioannou YA, Vanier MT, Ohno K, Ninomiya H.
  • Journal Title: J Cell Sci. 119 Pages: 2643-2653
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Chemical synthesis of the 3-sulfooxy-7-N-acetylglucosaminyl-24-amidated conjugates of 3β,7β-dihydroxy-5-cholen-24-oic acid, and related compounds : Unusual, major metabolites of bile acid in a patient with Niemann-Pick disease type C1. (2006)
  • Author(s): Iida T, Kakiyama G, Hibiya Y, Miyata S, Inoue T, Ohno K, Goto T, Mano N, Goto J, Nambara T, A F Hofmann.
  • Journal Title: Steroids 71 Pages: 18-29
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Chemical synthesis of the 3-sulfooxy-7-N-acetylglucosaminyl-24-amidated conjugates of 3β.7β-dihydroxy-5-cholen-24-oic acid, and related compounds : Unusual, major metabolites of bile acid in a patient with Niemann-Pick disease type C1. (2006)
  • Author(s): Iida T.et al.
  • Journal Title: steroids 71 Pages: 18-29
• [Journal Article] Activity of N-alkyl-β-valienamine sa a Chemical Chaperone for β-glucosidase Mutants Associated with Gaucher Disease. (2006)
  • Author(s): K Ohno, et al.
  • Journal Title: 10~<th> International Child Neurology Congress Pages: 135-138
• [Journal Article] Cholestrol depletion facilitates ubiquitylation of NPC1 and its association with SKD1/Vps4. (2006)
  • Author(s): Ohsaki Y. et al.
  • Journal Title: J Cell Sci, et al. 119 Pages: 2643-2653
• [Journal Article] Risk Factors for Fatality and Neurological Sequelae after Status Epilepticus in Children. (2005)
  • Author(s): Maegaki Y, et al.
  • Journal Title: Neuropediatrics 36 Pages: 186-192
• [Journal Article] Arthrogryposis multiplex congenita with callosal agenesis and dentato-olivary dysplasia. (2005)
  • Author(s): Saito Y, et al.
  • Journal Title: Brain Dev Dec17(Epub ahead of print)
• [Journal Article] Reduced sensitivity of Niemann-Pick C1-deficient cells to θ-toxin (perfringolysin O) : sequestration of toxin to raft-enriched membrane vesicles. (2004)
  • Author(s): Ohsaki Y, Sugimoto Y, Kaidoh T, Shimada Y, Ohno-Iwashita Y, Joanna P. Davies, Yiannis A. Ioannou, Ohno K, Ninomiya H
  • Journal Title: Histochem Cell Biol 121 Pages: 263-272
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Axonal dystrophy of dorsal root ganglion sensory neurons in a mouse model of Niemann-Pick disease type C.Experimental (2004)
  • Author(s): Ohara S, Ukita Y, Ninomiya H, Ohno K
  • Journal Title: Experimental Neurology 187 Pages: 289-298
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Degeneration of cholecystokinin- immunoreactive afferents to the VPL thalamus in a mouse model of Niemann-Pick disease type C. (2004)
  • Author(s): Ohara S, Ukita Y, Ninomiya H, Ohno K
  • Journal Title: Brain Research 1022 Pages: 244-246
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] N-Octyl-β-valienamine up-regulates activity of F213I mutant β-glucosidase in cultured cells : a potential chemical chaperone therapy for Gaucher disease. (2004)
  • Author(s): Lin H, Sugimoto Y, Ohsaki Y, Ninomiya H, Oka A, Taniguchi M, Ida H, Eto Y, Ogawa S, Matsuzaki Y, Sawa M, Inoue T, Higaki K, Nanba E, Ohno K, Suzuki Y
  • Journal Title: Biochim Biophys Acta 1689 Pages: 219-228
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Increased NPC1 mRNA in skin fibroblasts from Niemann-Pick disease type C patients. (2004)
  • Author(s): Yamamoto T, Feng J-H, Higaki K, Taniguchi M, Nanba E, Ninomiya H, Ohno K
  • Journal Title: Brain Dev 26 Pages: 245-250
  • NAID: 10020552645
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Reduced sensitivity of Niemann-Pick C1-deficient cells to θ-toxin (perfringolysin O) : sequestration of toxin to raft-enriched membrane vesicles. (2004)
  • Author(s): Ohsaki Y, Sugimoto Y, Kaidoh T, Shimada Y, Ohno-Iwashita Y, Joanna P.Davies, Yiannis A.Ioannou, Ohno K, Ninomiya H.
  • Journal Title: Histochem Cell Biol. 121 Pages: 263-272
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Axonal dystrophy of dorsal root ganglion sensory neurons in a mouse model of Niemann-Pick disease type C. (2004)
  • Author(s): Ohara S, Ukita Y, Ninomiya H, Ohno K.
  • Journal Title: Experimental Neurology. 187 Pages: 289-298
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Degeneration of cholecystokinin-immunoreactive afferents to the VPL thalamus in a mouse model of Niemann-Pick disease type C. (2004)
  • Author(s): Ohara S, Ukita Y, Ninomiya H, Ohno K.
  • Journal Title: Brain Research 1022 Pages: 244-246
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Increased NPC1 mRNA in skin fibroblasts from Niemann-Pick disease type C patients. (2004)
  • Author(s): Yamamoto T, Feng J-H, Higaki K, Taniguchi M, Nanba E, Ninomiya H, Ohno K.
  • Journal Title: Brain Dev. 26 Pages: 245-250
  • NAID: 10020552645
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] N-Octyl-β-valienamine up-regulates activity of F213I mutant β-glucosidase in cultured cells : a potential chemical chaperone therapy for Gaucher disease. (2004)
  • Author(s): Lin H, Sugimoto Y, Ohsaki Y, Ninomiya H, Oka A, Taniguchi M, Ida H, Eto Y, Ogawa S, Matsuzaki Y, Sawa M, Inoue T, Higaki K, Nanba E, Ohno K, Suzuki Y.
  • Journal Title: Biochim Biophys Acta. 1689 Pages: 219-228
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Novel TSC2 mutations and decreased expression of tuberin in cultured tumor cells with an insertion mutation (2004)
  • Author(s): Feng J-H, Yamamoto T, Nanba E, Ninomiya H. Oka A. Ohno K
  • Journal Title: HUMAN MUTATION #696(Online)
• [Journal Article] N-Octyl-β-valienamine up-regulates activity of F213I mutant β-glucosidase in cultured cells : a potential chemical chaperone therapy for Gaucher disease (2004)
  • Author(s): Lin H, Sugimoto Y, Ohsaki Y, Ninomiya H, Oka A, et al.
  • Journal Title: Biochim Biophys Acta 1689 Pages: 219-228
• [Journal Article] Axonal dystrophy of dorsal root ganglion sensory neurons in a mouse model of Niemann-Pick disease type C (2004)
  • Author(s): Ohara S, Ukita Y, Ninomiya H, Ohno K
  • Journal Title: Experimental Neurology 187 Pages: 289-298
• [Journal Article] Reduced sensitivity of Niemann-Pick C1-deficient cells to θ-toxin (perfringolysin O): sequestration of toxin to raft-enriched membrane vesicles (2004)
  • Author(s): Ohsaki Y, Sugimoto Y, Kaidoh T. Shimada Y, et al.
  • Journal Title: Histochem Cell Biol 121 Pages: 263-272
• [Journal Article] Increased NPC1 mRNA in skin fibroblasts from Niemann-Pick disease type C patients (2004)
  • Author(s): Yamamoto T, Feng J-H, Higaki K, Taniguchi M, Nanba E. Ninomiya H, Ohno K
  • Journal Title: Brain Dev 26 Pages: 245-250
  • NAID: 10020552645
• [Journal Article] Degeneration of cholecystokinin-immunoreactive afferents to the VPL thalamus in mouse model of Niemann-Pick disease type C (2004)
  • Author(s): Ohara S, Ukita Y, Ninomiya H, Ohno K
  • Journal Title: Brain Research 1022 Pages: 244-246
• [Book] 今日の小児治療指針 第14版「Niemann-Pick病」(大関武彦、古川漸、横田俊一郎 編) (2006)
  • Author(s): 大野耕策
  • Total Pages: 840
  • Publisher: 医学書院
  • Description: 「研究成果報告書概要(和文)」より
• [Book] Niemann-Pick diseases. In "Today's Therapy for Diseases of Children", 14^<th> edition. (Ohzeki T, Furukawa H, Yokota S (eds)) (2006)
  • Author(s): Kousaku Ohno
  • Total Pages: 840
  • Publisher: Igakushoin
  • Description: 「研究成果報告書概要(欧文)」より
• [Book] 今日の小児治療指針 第14版「Niemann-Pick病」(大関武彦, 古川漸, 横田俊一郎 編) (2006)
  • Author(s): 大野耕策
  • Total Pages: 840
  • Publisher: 医学書院