Functional analysis of HGF and its family member (HLP): from the basic to the potential application to neural diseases

HGF及其家族成员（HLP）的功能分析：从基础到在神经疾病中的潜在应用

• 批准号: 12680752
• 负责人: FUNAKOSHI Hiroshi
• 金额: $ 2.37万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12680752/
• 关键词: HGF; c-Met; HGF-like protein(HLP); Amyotrophic Lateral Sclerosis(ALS); Motor neurons; Transgenic mice; Gas6; Scavenger receptor; Hepatocyte Growth Factor(HGF); HGF-like protein(HLP); Ron; 神経栄養因子; 神経変性疾患; 運動ニューロン

Hepatocyte growth factor (HGF) was first purified as a mitogen for primary hepatocyte, and molecularly cloned in 1989. In 1995, our group identified HGF as a neurotrophic factor using primary hippocampal neurons. We here focused on the elucidation of the role of HGF both during neural development and in one of the neurodegenerative diseases, amyotrophic lateral sclerosis (ALS). During cortical development, HGF and c-Met/HGF receptor was regionally and sequentially regulated in a concerted fashion.In addition, HGF promoted neural migration and survival of cortical neurons, analyzed in primary dissociated neuroual culture and slice culture of cerebral cortex. These suggested the implication of HGF in cortical layer formation. To assess the role of HGF in ALS, we first generated trausgenic mice overexposing HGF in a neuron specific manner (HGF-Tg).HGF-Tg was mated with transgenic mice overrexpressing mutant SOD1 (ALS-Tg), transgenic mice model of ALS. Transgenic mice overexposing both HGF … More and mutant SOD1 (HGF/ALS-Tg). showed the attenuation of motoneuronal death, delayed onset, improved motor function and prolonged life span of ALS mice. Histological, western blotting, quantitative competitive RT-PCR analyzes together with primary culture study of neural cells revealed that HGF improves, ALS by attenuating induction of caspase-1 in motoneurons and by preventing reduction of gluamate transporter in astrocytes. These suggest the potential therapeutic activities of HGF on ALS and related disorders.HGF-like protein (HLP/MSP) is a family member of HGF HGF and its receptor Ron were expressed in the nervous system. However, function of HLP in the nervous system is not well understood. We found that HLP acts as a neurotrophic factor using chick dorsal ganglion explants. A chimeric receptor composed of extracellular and trausmembrane domains of Ron fused with immunoglobuline (Rou-Fc) eliminated activities of HLP on chick sensory ganglia, such as migration and neurite outgrowth promoting activities. This is the first evidence that HLP acts as a neurotrophic factor. Less

肝细胞生长因子（HGF）首先作为原代肝细胞的有丝分裂原被纯化，并于1989年进行分子克隆。1995年，我们的研究小组利用原代海马神经元将HGF鉴定为神经营养因子。我们在此重点阐明 HGF 在神经发育过程中以及在神经退行性疾病之一肌萎缩侧索硬化症 (ALS) 中的作用。在皮质发育过程中，HGF和c-Met/HGF受体以协调一致的方式进行区域性、顺序性调节。此外，在原代游离神经元培养物和大脑皮层切片培养物中分析，HGF促进皮质神经元的神经迁移和存活。这些表明 HGF 在皮质层形成中的意义。为了评估 HGF 在 ALS 中的作用，我们首先生成了以神经元特异性方式过度暴露 HGF 的致伤小鼠 (HGF-Tg)。HGF-Tg 与过度表达突变 SOD1 的转基因小鼠 (ALS-Tg) 交配，这是 ALS 转基因小鼠模型。过度暴露 HGF 和突变型 SOD1 (HGF/ALS-Tg) 的转基因小鼠。显示 ALS 小鼠运动神经元死亡减弱、发病延迟、运动功能改善和寿命延长。组织学、蛋白质印迹、定量竞争性 RT-PCR 分析以及神经细胞的原代培养研究表明，HGF 通过减弱运动神经元中 caspase-1 的诱导以及防止星形胶质细胞中谷氨酸转运蛋白的减少来改善 ALS。这些表明HGF对ALS及相关疾病具有潜在的治疗活性。HGF样蛋白(HLP/MSP)是HGF家族成员，HGF及其受体Ron在神经系统中表达。然而，HLP 在神经系统中的功能尚不清楚。我们使用鸡背神经节外植体发现 HLP 作为神经营养因子。由与免疫球蛋白 (Rou-Fc) 融合的 Ron 细胞外和膜内结构域组成的嵌合受体消除了 HLP 对小鸡感觉神经节的活性，例如迁移和神经突生长促进活性。这是 HLP 作为神经营养因子发挥作用的第一个证据。较少的

项目成果

Funakoshi H, Yonemasu T 他: "Identification of Gas6, a putative ligand for Sky and Axl receptor tyrosine kinases, as a novel neurotrophic factor for hippocampal neurons"J. Neurosci. Res.. (In press). (2002)

Funakoshi H、Yonemasu T 等人：“Sky 和 ​​Axl 受体酪氨酸激酶的推定配体 Gas6 作为海马神经元的新型神经营养因子的鉴定”J. Neurosci。

Funakoshi H, Nakamura T: "Identification of HGF-like protein as a novel neurotrophic factor for avian dorsal root ganglion sensory neurons"Biochem Biophys Res Commun.. 283, 3. 606-612 (2001)

Funakoshi H、Nakamura T：“鉴定 HGF 样蛋白作为禽类背根神经节感觉神经元的新型神经营养因子”Biochem Biophys Res Commun.. 283, 3. 606-612 (2001)

Tuzuki N, Miyazawa T 他: "Hepatocyte growth factor reduces the infarct volume after transient focal cerebral isehemia in rats"Neurol Res.. 23, 4. 417-424 (2001)

Tuzuki N、Miyazawa T 等人：“肝细胞生长因子可减少大鼠短暂局灶性脑缺血后的梗塞体积”Neurol Res.. 23, 4. 417-424 (2001)

S.Radaeva: "Unique epithelial cell production of hepatocyte growth factor/scatter factor by putative precancerous intestinal metaplasias and associated "intestinal-type" of biliary cancer chemically-induced in rat liver."Hepatology. 31. 1257-1265 (2000)

S.Radaeva：“通过假定的癌前肠化生和在大鼠肝脏中化学诱导的相关“肠型”胆道癌，上皮细胞产生独特的肝细胞生长因子/分散因子。”肝病学。

K.Matsumoto: "Hepatocyte growth factor in renal regeneration, renal disease, and potential therapeutics."Hypertension. 9. 395-402 (2000)

K.Matsumoto：“肝细胞生长因子在肾脏再生、肾脏疾病和潜在治疗中的作用。”高血压。