Studies on the pathophysiology and gene therapy for adrenoleukodystrophy using knock-out mice

敲除小鼠肾上腺脑白质营养不良的病理生理学和基因治疗研究

• 批准号: 08457191
• 负责人: YAMADA Takeshi
• 金额: $ 4.93万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08457191/
• 关键词: adrenoleukodystrophy; ALD; ALDP; knockout mice; very long chain fatty acid; beta-oxidation; VLACS; peroxisome; 遺伝子治療; レトロウイルスベクター

X-linked adrenoleukodystrophy (ALD) is caused by the mutation of ALD protein (ALDP) gene.Its principal biochemical abnormality is the accumulation of very long chain fatty acids (VLCFA) in tissues and body fluids, due to the impairment of beta-oxidatation in the peroxisome. We clarify its pathophysiology using the ALDP-deficient mice.VLCFA beta-oxidation in the ALD fibroblasts was not corrected by overxpression of VLACS only but done by overxpression of both VLACS and ALDP.Western blot analysis revealed that the VLACS protein was present in each tissue from the ALDP-deficient mouse. Liver peroxisomes were purified by Nycodenz gradient centrifugation. The VLACS protein was detected only in the peroxisomal fraction from the control mouse, while it was detected in the cytosol fraction as well as the peroxisomal fraction from the ALDP-deficient mouse. These results indicated that ALDP is involved in the transport of VLACS into the peroxisome and that VLACS can not catalyze VLCFA beta-oxidation unless localized in the peroxisome.

X连锁肾上腺脑白质营养不良（ALD）是由ALD蛋白（ALDP）基因突变引起的。其主要生化异常是由于β-氧化受损导致极长链脂肪酸（VLCFA）在组织和体液中积累在过氧化物酶体中。我们使用 ALDP 缺陷小鼠阐明其病理生理学。ALD 成纤维细胞中的 VLCFA β-氧化不能仅通过 VLACS 的过表达来纠正，而是通过 VLACS 和 ALDP 的过表达来纠正。Western blot 分析显示，每个组织中都存在 VLACS 蛋白来自 ALDP 缺陷小鼠。通过 Nycodenz 梯度离心纯化肝脏过氧化物酶体。仅在对照小鼠的过氧化物酶体级分中检测到 VLACS 蛋白，而在 ALDP 缺陷型小鼠的胞质级分和过氧化物酶体级分中均检测到 VLACS 蛋白。这些结果表明，ALDP 参与 VLACS 向过氧化物酶体的转运，并且 VLACS 不能催化 VLCFA β-氧化，除非位于过氧化物酶体中。

项目成果

Yamada Takeshi: "Protease inhibitors suppress the degradation of mutant adrenoleukodystrophy proteins but do not correct impairment of very long chain fatty acid metabolism in adrenoleukodystrophy fibroblasts" Neurochemical Research. (in press).

Yamada Takeshi：“蛋白酶抑制剂抑制突变型肾上腺脑白质营养不良蛋白的降解，但不能纠正肾上腺脑白质营养不良成纤维细胞中极长链脂肪酸代谢的损害”神经化学研究。

Kobayashi Takuro: "Adrenoleukodystrophy protein-deficient mice represent abnormality of very long chain fatty acid metabolism" Biochem Biophys Res Commun. 232. 631-636 (1977)

小林拓郎：“肾上腺脑白质营养不良蛋白缺陷小鼠代表极长链脂肪酸代谢异常”Biochem Biophys Res Commun。

Kobayashi T,Shinnoh N,Kondo A,Yamada T: "Adrenoleukodystrophy protein-deficient mice represent abnormality of very long chain fatty acid metabolism" Biochem Biophys Res Commun. 232. 631-636 (1997)

Kobayashi T、Shinnoh N、Kondo A、Yamada T：“肾上腺脑白质营养不良蛋白缺陷小鼠代表极长链脂肪酸代谢异常”Biochem Biophys Res Commun。

Yamada Takeshi: "Protease inhibitor suppress the degradation of mutant adrenoleuko-dystrophy proteins but do not correct irmpaiment of very long chain fatty acid metabolism in adrenoleukodystrophy fibroblasts" Neurochemical Research. 22. 233-237 (1997)

Yamada Takeshi：“蛋白酶抑制剂抑制突变型肾上腺脑白质营养不良蛋白的降解，但不能纠正肾上腺脑白质营养不良成纤维细胞中极长链脂肪酸代谢的损伤”神经化学研究。

Yamada Takeshi: "Protease inhibitor suppress the degradation of mutant adrenoleukodystrophy proteins but do not correct impairment of very long chain fatty acid metabolism in adrenoleukodystrophy fibroblasts" Neurochemical Research. 22. 233-237 (1997)

Yamada Takeshi：“蛋白酶抑制剂抑制突变型肾上腺脑白质营养不良蛋白的降解，但不能纠正肾上腺脑白质营养不良成纤维细胞中极长链脂肪酸代谢的损害”神经化学研究。