Investigation of a new molecular target-based treatment of caneer-drug resistant pediatric solid tumors

癌症耐药儿童实体瘤新分子靶向治疗的研究

• 批准号: 15591098
• 负责人: KAMIJO Takehiko
• 金额: $ 2.24万
• 依托单位: Shinshu University Hospital
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591098/
• 关键词: neuroblastoma; MDM2; p53; Apoptosis; localization; mitochondria; Noxa; Doxorubicin

Neuroblastoma (NB) is the most common pediatric solid malignant tumor derived from the sympathetic nervous system. Unlike the improvements in survival in many childhood malignancies, high-risk neumblastoma is still one of the most difficult tumors to cure, with only 30% long-term survival despite intensive multi-modal therapy. New treatments and a better understanding of drug resistance mechanisms are required for improvement of the survival rates. A noteworthy finding of neuroblastoma research is that mutations of p53 tumor suppressor have been reported less than 2% of NBs out of 340 tested (Tweddle DA et al., Cancer Res 61 : 8-13, 2001). Instead of mutation, cytoplasmic sequestration of p53 has been proposed as an alternative mechanism of inactivation in NB cells. The sequestration was first reported in frozen tumor samples using immuno-histodremical techniques (Moll UM et al., PNAS 92 : 4407-11, 1995) and later in NB oil lines by immuno-fluorescence and cell fractionation experiment … More s (Moll UM et al., MCB 16 : 1126-37, 1996). However, the p53 localization before and after DNA damage stimulation, p53 stabilization and activation by phosphorylation, binding to the p53-binding consensus sequences in vitro and in vivo, and up-regulation of p53-dowmstream molecules in NB cells have remained to be determined.Therefore, we employed a panel of cell lines to determine whether the p53-dependent cell death in NB cells is caused by apoptotic cellular function, and we further studied the molecular mechanism of apoptosis induced via p53-dependent pathway. We provide lines of evidence that a p53-dependent stress, doxorubicin (Doxo) administration, causes accumulation of p53 in nucleus of NB cells and phosphorylation of several serine residues (serine 15, serine 20, and serine 46) in both Doxo-sensitive and -resistant NB cell lines. Up-regulation of a p53-downstream molecule, p2lCip 1/Waf1 and accumulation of Noxa, a pro-apoptotic Bcl-2 family molecule, in mitochondria are observed in only Doxo-sensitive NB cells. Mitochondrial dysfunction including cytochrome c release and membrane potential dysregulation was caused in the sensitive cells, resulting in the activation of the intrinsic caspase pathway. However, the accumulation in nucleus and phosphorylation of p53 were inert to induce the Noxa accumulation and the mitochondrial dysfunction in p53-resistant NB cells. Surprisingly, inactivation of the p53 by over-expression of MDM2 resulted in the canceling of up-regulation of Noxa in mitochondria and reduced the apoptotic cell death. Taken together, MDM2/p53 pathway seems to play an important role in NB cells by Noxa regulation in mitochondria. Less

神经母细胞瘤（NB）是最常见的小儿交感神经系统实体恶性肿瘤。与许多儿童恶性肿瘤的生存率提高不同，高危成神经细胞瘤仍然是最难治愈的肿瘤之一，尽管采用了强化的多模式治疗，但长期生存率仅为30%。需要新的治疗方法和更好地了解耐药机制，以提高生存率。神经母细胞瘤研究的一个值得注意的发现是，在340个测试的NB中，已经报道了小于2%的p53肿瘤抑制基因突变（Tweddle DA等人，Cancer Res 61：8-13，2001）。代替突变，p53的细胞质隔离已被提出作为NB细胞中失活的替代机制。使用免疫组织化学技术在冷冻肿瘤样品中首次报道了隔离（Moll UM等人，PNAS 92：4407-11，1995），以及后来通过免疫荧光和细胞分级实验在NB油系中的应用 ...更多信息 s（Moll UM等人，MCB 16：1126-37，1996）。然而，在DNA损伤刺激之前和之后的p53定位、通过磷酸化的p53稳定和活化、在体外和体内与p53结合共有序列的结合以及NB细胞中p53下游分子的上调仍有待确定。我们使用一组细胞系来确定NB细胞中的p53依赖性细胞死亡是否由凋亡细胞功能引起，并进一步研究了p53依赖性途径诱导细胞凋亡的分子机制。我们提供的证据表明，p53依赖的压力，阿霉素（Doxo）管理，导致积累的p53在核的NB细胞和磷酸化的几个丝氨酸残基（丝氨酸15，丝氨酸20，和丝氨酸46）在这两个Doxo敏感和耐NB细胞系。仅在Doxo敏感的NB细胞中观察到p53下游分子p21 Cip 1/Waf 1的上调和线粒体中促凋亡Bcl-2家族分子Noxa的积累。在敏感细胞中引起线粒体功能障碍，包括细胞色素c释放和膜电位失调，导致内源性caspase途径激活。然而，在p53抗性NB细胞中，p53在细胞核中的积聚和磷酸化对诱导Noxa积聚和线粒体功能障碍是惰性的。令人惊讶的是，通过MDM 2的过表达使p53失活导致线粒体中Noxa的上调被取消，并减少了凋亡性细胞死亡。综上所述，MDM 2/p53通路似乎通过线粒体中的Noxa调节在NB细胞中发挥重要作用。少

项目成果

ST1571 inhibits growth and adhesion of human mast cells in culture.

ST1571 抑制培养物中人类肥大细胞的生长和粘附。

• 发表时间: 2003
• 期刊: J Leukoc Biol. 74
• 作者: Takeuchi K;Koike K;Kamijo T;他10名
• 通讯作者: 他10名

ARF Tumor Suppressor Induces Mitochondria-dependent Apoptosis by Modulation of Mitochondrial Bcl-2 Family Proteins*

• DOI: 10.1074/jbc.m300510200
• 发表时间: 2003-07
• 期刊: Journal of Biological Chemistry
• 影响因子: 4.8
• 作者: Y. Nakazawa;T. Kamijo;K. Koike;T. Noda

STI571 inhibits growth and adhesion of human mast cells in culture

STI571 抑制培养物中人类肥大细胞的生长和粘附

• DOI: 10.1189/jlb.0602284
• 发表时间: 2003
• 期刊: Journal of Leukocyte Biology
• 影响因子: 5.5
• 作者: K. Takeuchi;K. Koike;T. Kamijo;S. Ishida;Y. Nakazawa;Y. Kurokawa;K. Sakashita;T. Kinoshita;Shigeyuki Matsuzawa;M. Shiohara;T. Yamashita;M. Nakajima;A. Komiyama
• 通讯作者: A. Komiyama

Successful unrelated cord blood transplantation using a reduced-intensity conditioning regimen in a 6-month-old infant with congenital neutropenia complicated by severe pneumonia.

对一名患有先天性中性粒细胞减少症并发严重肺炎的 6 个月大婴儿，使用降低强度调理方案成功进行无关脐带血移植。

• 发表时间: 2004
• 期刊: Int J Hematol. 80(3)
• 作者: Nakazawa Y;Sakashita K;Kinoshita M;Saida K;Shigemura T;Yanagisawa R;Shikama N;Kamijo T;Koike K.
• 通讯作者: Koike K.

Tanaka M, Kamijo T et al.: "Specific Autoantibodies to Platelet Glycoproteins in Epstein-Barr Virus-Associated Immune Thrombocytopenia"Int J Hematol. 78. 168-170 (2003)

Tanaka M、Kamijo T 等人：“Epstein-Barr 病毒相关免疫性血小板减少症中血小板糖蛋白的特异性自身抗体”Int J Hematol。