The study of control technique of interspecies signal transfer

种间信号传递控制技术研究

• 批准号: 08559002
• 负责人: TANAKA Tomoo
• 金额: $ 10.69万
• 依托单位: Tokai University (1997-1998)University of Tsukuba (1996)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08559002/
• 关键词: 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 … More 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

为了检测种间信号传递的物质，我们使用了针对人脑和寄生虫原生动物特异性的单克隆抗体，Cruzi锥虫（Wood。J.N.EtAl。，Nature，Nature，1982）。我们筛选了人体组织和t.cruzi cDNA文库，并获得了几个新基因。其中，北印迹选择了在人脑，心脏肌肉和t.cruzi中表达的新型基因大体，最初由单克隆抗体检测到。我们得到了以下有关该基因及其蛋白质的结构和功能表征：1。 Bradeion在成人人脑中被表示为两种类型（称为A和Beta）。它的氨基酸比对具有生长马和细胞因子受体的基序。它也被认为是具有跨膜区域和ATP/GTP结合位点的孤儿受体2。 Bradejon在Northern印迹中的信号仅在成年人的大脑和心脏（脑的10％）上显示，并且在任何其他组织和胚胎阶段都没有信号。3。 Bradejon的β型仅在小鼠大脑中更为表达（对人类大型β的身份为94％）。该基因位于小鼠VH染色体中，并以3个外显子和2个覆盖Ca的内含子的形式存在。 17 kb.4。 Bradeion Alpha位于线粒体，核核细胞胞浆菌M.5。凋亡发生在人类培养的癌细胞中，这些细胞被翻译成6。 Bradeion基因也在有色癌和恶性黑色素瘤的人培养的癌细胞中表达。在人类有色癌和恶性黑色素瘤患者的样品中也观察到了这种表达。7。当翻译反义核酶时，观察到了显着的生长和转化。基于这项研究的新医学和医疗机械现在也在开发。此外，已经执行了使用C.Elegans或Mouse等动物模型及其应用于人类的动物模型发现相关物质的试验。但是，该研究受到大脑系统组织和细胞所具有的物种和个体差异的限制。因此，需要建立使用人类文化细胞的方法。从观点考虑人类和其他生物的共同进化或对环境的适应，即“寄生虫”，我们的研究表明，我们的研究证明了细胞年龄的新控制因素及其功能分析的发现。根据这些发现，我们开始了以下研究，将Bradeion应用于现在的全球和全球行业的种子：1。新的人脑孤儿受体的功能分析Bradeion作为有用的物理激活剂2。分析神经元如何非常复杂和高级结构和神经元的程序性细胞死亡的分析。3。用于控制神经元死亡，早期癌症诊断和癌症基因治疗的技术和医学机械的开发。较少的

项目成果

Tanaka T: "Differential Diagnosis of Trpanosoma cruzi and T.rangeli Infection by PCR of Cysteine Proteinase Genes" J Jpn Assoc Inf Dis. 71・9. 903-909 (1997)

Tanaka T：“通过半胱氨酸蛋白酶基因的PCR对克氏锥虫和兰格利锥虫感染进行鉴别诊断”J Jpn Assoc Inf Dis. 71・909。

Tanaka M.,et al: "Schistosoma mansoni Genome Project Network" Parasitology Today. (in press). (1996)

Tanaka M.等人：“曼氏血吸虫基因组计划网络”今日寄生虫学。

Tanaka,M.,et al: "Proceedings of Schistozoma genome Project/96" Mem.Inst.Oswald Cruz.(in press). (1996)

Tanaka,M.,et al:“裂殖虫基因组计划/96”Mem.Inst.Oswald Cruz.（正在出版）。

Tanaka M: "The Triplet Repcat(Alanine;GCT)in Ribosomal Protein L14 of Immortalized Human Endothelial Cell Line(t-HUEA)." Biochemical and Biophysical Research Communications. (in press). (1998)

Tanaka M：“永生化人内皮细胞系 (t-HUEA) 核糖体蛋白 L14 中的三联体 Repcat（丙氨酸；GCT）。”

Tanaka T: "Analysis of Expressed Sequence Tags (ESTs) of Parasitic Protozoa Entamoeba histolytica" Biochemical and Biophysical Research Communications. 236・3. 611-615 (1997)

田中 T：“寄生原生动物溶组织阿米巴的表达序列标签（EST）分析”生物化学和生物物理研究通讯236・3（1997）。