Grant-in-Aid for Scientific Research (A)
|Allocation Type||Single-year Grants|
|Research Institution||Tokai University(1997-1998)|
University of Tsukuba(1996-1997)
TANAKA Tomoo(1997-1998) Tokai University, School of Medicine, Research Associate, 医学部, 助手 (50192175)
田中 真奈実(1996-1997) 筑波大, 基礎医学系, 講師 (80188341)
MURAKAMI Hiroshi Nippon Ham, Ltd., Central Institute, Researcher., 中央研究所, 研究員
MITSUI Youji Ministry of International Trade and Industry, Agency of Industrial Science and T, 生命工学工業技術研究所, 首席研究官
TANAKA Manami Ministry of International Trade and Industry, Agency of Industrial Science and T, 生命工学工業技術研究所, 主任研究官
田中 真奈美 通商産業省工業技術院, 生命工学工業技術研究所, 主任研究官 (80188341)
田中 朝雄 東海大学, 医学部, 助手 (50192175)
|Project Period (FY)
1996 – 1998
Completed(Fiscal Year 1998)
|Budget Amount *help
¥16,700,000 (Direct Cost : ¥16,700,000)
Fiscal Year 1998 : ¥3,900,000 (Direct Cost : ¥3,900,000)
Fiscal Year 1997 : ¥5,900,000 (Direct Cost : ¥5,900,000)
Fiscal Year 1996 : ¥6,900,000 (Direct Cost : ¥6,900,000)
|Keywords||interspecies signal transfer / Bradeion / orphan receptor / apoptosis / antisense-ribozyme / cancer-related gene / gene diagnosis / gene therapy / 脳・心筋特異遺伝子 / インターロイキン受容体 / CE5B3 / Green Fluorescent Protein / 融合蛋白質 / シャガス病 / クルーズ・トリパノソーマ / エンドセリン / 腸管神経節|
To detect the substance of interspecies signal transfer, we used the monoclonal antibody specific to human brain and parasitic protozoa, Trypanosoma cruzi (Wood. J.N.et al., Nature, 1982). We screened human tissues and T.cruzi cDNA library, and obtained several novel genes. Among them, Northern blot selected novel gene Bradeion expressing in human brain, cardiac muscle and T.cruzi, which were originally detected by the monoclonal antibody. We got the following structural and functional characterization about this gene and its protein :
1. Bradeion was expressed as two types (called as a and beta) in adult human brain. The amino acid alignment of it has the motif of growth hormone and cytokine receptor. It was also considered as an orphan receptor with transmembrane region and ATP/GTP binding site.
2. The signal of Bradejon in Northern blot was shown only on adult human brain and heart (10% of brain), and no signal in any other tissues and embryonic stage.
3. Only beta-type of Bradejon was
expressed in mouse brain (94% identity to human Bradeion beta). This gene is located in mouse chromosome VH and exists as 3 exons and 2 introns covering ca. 17 kb.
4. Bradeion alpha was located in mitochondria and Bradeion beta in perinuclear cytoplasm.
5. Apoptosis occurs in human cultured cancer cell in which Bradeion was transfected.
6. Bradeion gene was also expressed in human cultured cancer cells of colorectal cancer and malignant melanoma. This expression was also observed in the samples of human colorecial cancer and malignant melanoma patients.
7. The remarkable decrease of growth and the transformation in cultured colorectal cancer cell were observed when antisense ribozyme was transfected.
The molecular pathology of neural degenerative disease like Alzheimer disease and Huntington disease has been studied to control the age and prevention of death of neural cell. New medicine and medical machinery based on this study are also developing now. Furthermore, the trial finding the related substance using animal models like C.elegans or mouse and its application to human have been performed. However, the study is limited by the species and individual differences, which the tissue and the cell of the cerebral system have. The establishment of the methodology to use human culture cell is therefore needed. Bradeion is quite a novel substance concerning to flue survival (death) of human neuron gained by approach from the point of view considering co-evolution of human and other organisms or adaptation of organism to environment, i. e., "parasitism".
Consequently, our study demonstrated the discovery of new control factor of cell age, Bradeion and its functional analysis. According to these findings, we started the following studies to apply Bradeion as the seed of a new and global industry now :
1. Analysis of function of a new human brain orphan-receptor, Bradeion as useful physiological activator.
2. Analysis of the mechanism how neurons contract very complex and high-grade structure and the programmed cell death of neuron happens.
3. Development of technology and medical machinery for control of neural death, diagnosis of early cancer and gene therapy of cancer. Less