DEVELOPMENT OF THE POSITIVE-FEEDBACK TRANSCRIPTIONAL/REGULATION SYSTEM FOR A DNA VACCINE VECTOR

• Project/Area Number: 17590425
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Virology
• Research Institution: YOKOHAMA CITY UNIVERSITY SCHOOL OF MEDICINE
• Principal Investigator: SASAKI Shin YOKOHAMA CITY UNIV., SCH. OF MED, ASSOCIATE PROFESSOR, 医学部・客員準教授 (40342901)
• Co-Investigator(Kenkyū-buntansha): TAKESHITA Fumihiko YOKOHAMA CITY UNIV., SCH. OF MED, ASSOCIATE PROFESSOR, 医学部, 準教授 (60333572)
• Project Period (FY): 2005 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥3,500,000 (Direct Cost: ¥3,500,000); Fiscal Year 2006: ¥1,100,000 (Direct Cost: ¥1,100,000); Fiscal Year 2005: ¥2,400,000 (Direct Cost: ¥2,400,000)
• Keywords: DNA VACCINE / TRANSCRUPTION FACTOR / GENE REGULATION

Research Abstract

MAINTAINANCE OF HIGH-LEVEL TRANSGENE EXPRESSION IS THE MAIN CHALLENGE IN CURRENT GENE THERAPY AND GENE-MEDIATED VACCINES. ALTHOUGH THE CYTOMEGALOVIRUS (CMV) PROMOTER/ENHANCER OR ITS DERIVATIVE THE CAG PROMOTER HAS BEEN HARNESSED IN CURRENT GENE THERAPY AND GENE-MEDIATED VACCINE VECTORS, TRANSGENE EXPRESSION BY THESE VECTORS IS OFTEN TRANSIENT AND REMAINS AT SUBOPTIMAL LEVELS DUE TO UNDEFINED MECHANISMS, POSSIBLY INCLUDING THE SHORTAGE OF TRANSCRIPTIONAL MACHINERY. TO OVERCOME THIS DRAWBACK, WE DESIGNED A NOVEL TRANSCRIPTIONAL CONTROL SYSTEM, DESIGNATED HERE AS TRANSCRIPTION FACTOR SUPERCHARGING PROMOTER SYSTEM, IN WHICH TRANSGENE EXPRESSION IS REGULATED BY THE POSITIVE FEEDBACK CIRCUIT CONSISTING OF CIS-AND TRANS-ACTING ELEMENTS OF GENE EXPRESSIONMACHINERY. AMONG COMBINATIOINS OF THESE ELEMENTS, A PLASMID COMPOSED OF A TARGET GENE EXPRESSION CASSETTE DRIVEN BY THE CHIMERIC CMV PROMOTER CONTAINING REPETITIVE 12-0-TETRADECANOYLPHORBOL-13-ACETATE-RESPONSIVE ELEMENTS AS CIS-ACTING ELEMENTS (CMV-TTT) AND EXPRESSION CASSETTES FOR C-FOS AND C-JUN GENES AS TRANS-ACTING ELEMENTS FACILITATED HIGH AND LONG-TERM (>10 MONTHS) EXPRESSION OF A TRANSGENE AFTER IYS INTRAMUSCULAR ELECTROPORATION-MEDIATED DELIVERY IN MICE.. SINCE HUMAN SECRETORY ALKALINE PHOSPHATASE WAS USED AS A REPORTER, IT WAS SUGGESTED THAT THE IMMUNE EVASION MECHANISM ELICITED BY THE CMV-TTT AND/OR C-FOS/C-JUN EXPRESSION ALSO CONTRIBUTED TO THE SUSTAINED EXPRESSION IN MICE.. OUR STRATEGY MAY OPEN A NEW AVENUE FOR A GENE THERAPY THAT INVOLVES LIFELONG SUPPLEMENTATION OF A DEFICIENT PROTEIN THAT COULD BE TARGETED THE HOST' S IMMUNE SYSTEM.

Research Products

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  • Journal Title: Endocrinology (in press)
  • Description: 「研究成果報告書概要(和文)」より
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• [Journal Article] Suppression of iodide (2007)
  • Author(s): Yamazaki K, Suzuki K, Yamada E, Yamada T, Takeshita F, Matsumoto M, Mitsuhashi T, Obara T, Takano K, Sato K.
  • Journal Title: Endocrinology (in press)
  • Description: 「研究成果報告書概要(欧文)」より
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  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Suppression of iodide uptade and thyroid hormone synthesis with stimulation of the type I interferon system by double-stranded RNA (dsRNA) in cultured human thyroid follicles. (2007)
  • Author(s): Yamazaki K, Suzuki K, Yamada E, Yamada T, Takeshita F, Matsumoto M, Mitsuhashi T, Obara T, Takano K, Sato K
  • Journal Title: Endocrinology (in press)
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  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Multivalent DNA vaccine protects mice against pulmonary infection caused by Pseudomonas aeruginosa (2006)
  • Author(s): Saha S, Takeshita F, Sasaki S, Matsuda T, Tanaka T, Tozuka M, Takase K, Matsumoto T, Okuda K, Ishii N, Yamaguchi K, Xin KQ, Klinman DM, Okuda K
  • Journal Title: Vaccine 24 Pages: 6240-6249
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  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] A fused gene of nucleoprotein (NP) and herpes simplex virus genes (VP22) induces highly protective immunity against different subtypes of influenza virus. (2006)
  • Author(s): Saha S, Yoshida S, Ohba K, Matsui K, Matsuda T, Takeshita F, Umeda K, Tamura Y, Okuda K, Klinman D, Xin KQ, Okuda K.
  • Journal Title: Virology 35 Pages: 48-57
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Multivalent DNA vaccine protects mice against pulmonary infection caused by Pseudomonas aeruginosa. (2006)
  • Author(s): Saha S, Takeshita F, Sasaki S, Matsuda T, Tanaka T, Tozuka M, Takase K, Matsumoto T, Okuda K, Ishii N, Yamaguchi K, Xin KQ, Klinman DM, Okuda K.
  • Journal Title: Vaccine 24 Pages: 6240-6249
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Activator protein 1-mediated transcriptional regulation strategy sustains long-term expression of a xenogeneic gene product in vivo : An implication for gene therapy targeting congenital protein deficiencies. (2006)
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  • Journal Title: Int.J.Mol.Med. 18 Pages: 289-297
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• [Journal Article] TLR-adaptor molecules enhance DNA-raised adaptive immune responses against flu and tumor challenge through the activation of innate immunity. (2006)
  • Author(s): Takeshita F, Tanaka T, Matsuda T, Tozuka M, Kobiyama K, Saha S, Matsui K, Ishii KJ, Coban C, Akira S, Ishii N, Suzuki K, Klinman DM, Okuda K, Sasaki S.
  • Journal Title: J.Virol. 80 Pages: 6218-6224
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• [Journal Article] Double stranded B-DNA activates type-I IFNs gene program via IPS-1/TBK1 pathway. (2006)
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  • Journal Title: J.Invest.Dermatol 125 Pages: 1156-1156
• [Journal Article] Interleukin-1 receptor-associated kinase-1 plays an essential role for Toll-like receptor (TLR)7- and TLR9-mediated interferon-alpha induction (2005)
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