GENETIC STUDY OF A CHILDHOOD DISEASE AFFECTNG INTRACELLULAR CHOLESTEROL TRANSPORT

影响细胞内胆固醇转运的儿童疾病的遗传学研究

• 批准号: 04670597
• 负责人: OHNO Kousaku
• 金额: $ 1.41万
• 依托单位: TOTTORI UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04670597/
• 关键词: Niemann-Pick Desease; cholesterol transport; selectable markers; gene cloning; immotalized cell; 細胞内転送; 染色体マッピング

Basic defectin type C Niemann-Pick disease is unknown. As an attempt to isolate the gene defectivein type C Niemann-Pick Disease, we have established an immortalized 3T3 cell line from sphingomelinosis mouse and found the cells show the same abnormalitis found in fibroblasts from patients with type C Niemann-Pick disease. In course of studies to characterizecholesterol metabolism in type C Niemann-Pick disease, we have fund fibroblasts from the patients have an increasedde novo pathway of cholesterol biosynthesis. When cells ara exposed to cholesterol biosynthetic inhibitors, they showed abnormal sensitivities. They showed marked hypersensitivity to cytotoxic effect of vitamin D3. The drug should be very useful to enrich and isolate a few cells restored cholesterol metabolism after transfecting human normal cDNA or genomic DNA.In addition, by transfer of a human chromosome into 3T3 cell line from sphingomyelinosis mouse, we have found a human chromosome 18 restore the cholesterol metabolism. BY Southern analysis, the gene locus is assigned to distal portion of the chromosome. These results suggest that the gene defective in type C Niemann-Pick disease could be isolated by expresstion cloning using the mouse cell line and the selectable drug or by positional cloning

C型尼曼-匹克病的基础缺陷尚不清楚。为分离C型尼曼-匹克病基因缺陷型，我们建立了一个鞘氨醇髓鞘病小鼠永生化3 T3细胞系，发现该细胞具有与C型尼曼-匹克病患者成纤维细胞相同的异常。在研究C型尼曼-匹克病的胆固醇代谢特征的过程中，我们发现患者的成纤维细胞具有增加的胆固醇生物合成的从头途径。当细胞暴露于胆固醇生物合成抑制剂时，它们表现出异常的敏感性。它们对维生素D3的细胞毒性作用表现出明显的超敏反应。此外，我们还将人染色体转入鞘磷脂病小鼠3 T3细胞系，发现人18号染色体具有恢复胆固醇代谢的作用。经Southern杂交分析，该基因定位于染色体的远端。这些结果表明，C型尼曼-匹克病的缺陷基因可以通过表达克隆和选择性药物或定位克隆来分离

项目成果

K.Ohno: "A cell line derived from sphingomyelinosis mouse shows alterations in intracellular cholesterol metabolism similar to those in type C Niemann-Pick disease." Cell Structure and Function. 17. 229-235 (1992)

K.Ohno：“源自鞘磷脂病小鼠的细胞系显示出与 C 型尼曼-匹克病相似的细胞内胆固醇代谢变化。”

K.Ohno.: "Altered sensitivities to potential inhibitors of cholesterol biosynthesis in Niemann-Pick type C fibroplasts." Cell Structure and Funotion. 18. 231-240 (1993)

K.Ohno.：“改变了 Niemann-Pick C 型纤维细胞中胆固醇生物合成潜在抑制剂的敏感性。”

A.Kurimasa: "Restoration of the cholesterol metabolism in 3T3 cell line derived from the sphinogomyelinosis mouse (spm/spm)by transfer of a human chromosome 18." Human Genetics. 92. 157-162 (1993)

A.Kurimasa：“通过转移人类 18 号染色体，恢复来自鞘磷脂病小鼠 (spm/spm) 的 3T3 细胞系中的胆固醇代谢。”

S.Akaboshi: "A hypoxanthine-guanine phosphoribosyl transferase deficient mutant from a cell line derived from a mouse model with a Niemaun-pick disease type C." Yonago Acta medica. 35. 221-230 (1992)

S.Akaboshi：“次黄嘌呤鸟嘌呤磷酸核糖基转移酶缺陷突变体，来自患有 C 型 Niemaun-pick 病的小鼠模型衍生的细胞系。”

K.Ohno: "A cell line derived from sphingomyelinosis mouse shows alterations in intracellular cholesterol metabolism similar to those in type C Niemann-Pick disease" Cell Structureand Function. 17. 229-235 (1992)

K.Ohno：“源自鞘磷脂病小鼠的细胞系显示出与 C 型尼曼-匹克病相似的细胞内胆固醇代谢变化”《细胞结构和功能》。