Development of safety vector for the gene therapy of ribosomal protein S19 deficient Diamond-Blackfan anemia

核糖体蛋白S19缺陷型Diamond-Blackfan贫血基因治疗安全载体的开发

• 批准号: 15591025
• 负责人: HAMAGUCHI Isao
• 金额: $ 2.24万
• 依托单位: National Institute of Infectious Diseases (2004)Keio University (2003)
• 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-15591025/
• 关键词: Diamond-Blackfan anemia; RPS19; hematopoietic stem cells; knockout mouse; siRNA; Red cell production; Diamond-Blackfan anemia; レンチウイルスベクター; ハイブリッドベクター

We have developed new lentiviral vector for the gene therapy of ribosomal protein S19(RPS19) deficient Diamond-Blackfan anemia(DBA). Since the strong transgene expression activity was required for this vector, we developed hybrid vector between lentiviral backborn and retrovirus U3 region in the LTR(long term repeat). Using this vector high transduction efficiency was observed in the DBA patient hematopoietic stem cells (Mol.Ther., 2003).In order to analyze the mechanisms for abnormal differentiation of erythroid leneage cells in DBA, RPS19-knock out mice were generated in the corraboration study with Niklas Dahl. In this mice we could not detect the typical phenotype of anemia correlated to human disease. It is suggested that the function of mouse RPS19 is different from human RPS19 (Mol.Cell.Biol., 2004).For further analyze of human RPS19 function, we developed lentiviral vector containing siRNA-expression cassette against RPS19. When we transduced siRNA gene against RPS19 in human cord blood CD34+ cells, the growth and differentiation of erythroid leneage cells were disturbed in the transduced cells (Blood, 2005).Together with these results, we have revealed the function of RPS19 and developed the stem cell model for DBA using siRNA vector. By using this model we would like to analyze the whole mechanisms of DBA, and develop the new therapy.

我们开发了新的慢病毒载体，用于核糖体蛋白S19（RPS19）缺陷型Diamond-Blackfan贫血（DBA）的基因治疗。由于该载体需要强大的转基因表达活性，我们开发了慢病毒backborn和LTR（长期重复）中逆转录病毒U3区域的杂合载体。使用该载体在DBA患者造血干细胞中观察到高转导效率（Mol.Ther.，2003）。为了分析DBA中红系细胞异常分化的机制，在与Niklas Dahl的验证研究中产生了RPS19敲除小鼠。在这些小鼠中，我们无法检测到与人类疾病相关的典型贫血表型。表明小鼠RPS19的功能与人RPS19不同(Mol.Cell.Biol., 2004)。为了进一步分析人RPS19的功能，我们开发了含有针对RPS19的siRNA表达盒的慢病毒载体。当我们在人脐带血CD34+细胞中转导针对RPS19的siRNA基因时，转导细胞中红系细胞的生长和分化受到干扰（Blood，2005）。结合这些结果，我们揭示了RPS19的功能，并使用siRNA载体开发了DBA干细胞模型。通过使用这个模型，我们希望分析DBA的整个机制，并开发新的疗法。

项目成果

Proliferation deficiency of multipotent hematopoietic progenitors in ribosal protein S19 (RPS19)-deficient diamond-Blackfan anemia improves following RPS19 gene transfer

RPS19基因转移后核糖蛋白S19（RPS19）缺陷的钻石-Blackfan贫血症改善多能造血祖细胞的增殖缺陷

• 发表时间: 2003
• 期刊: Mol Ther. 5
• 作者: Matsson H;Davey E;Draptchinskaia N;Hamaguchi I;Ooka A;Leveen Per;Forsberg E;Karlsson S;Dahl N;Isao Hamaguchi et al.
• 通讯作者: Isao Hamaguchi et al.

Proliferation deficiency of multipotent hematopoietic progenitors in ribosomal protein S19 (RPS19)-deficient Diamond-Blackfan anemia improves following RPS19 gene transfer

• DOI: 10.1016/s1525-0016(03)00091-1
• 发表时间: 2003-05-01
• 期刊: MOLECULAR THERAPY
• 影响因子: 12.4
• 作者: Hamaguchi, I;Flygare, J;Karlsson, S
• 通讯作者: Karlsson, S

Deficiency of ribosomal protein S19 in CD34+ cells generated by siRNA blocks erythroid development and mimics defects seen in Diamond-Blackfan anemia

• DOI: 10.1182/blood-2004-08-3115
• 发表时间: 2005-06-15
• 期刊: BLOOD
• 影响因子: 20.3
• 作者: Flygare, J;Kiefer, T;Karlsson, S
• 通讯作者: Karlsson, S

Hamaguchi I., Flygare J.et al.: "Proliferation deficiency of multipotent hematopoietic progenitors in ribosomal protein S19 (RPS19)-deficient Diamond-Blackfan anemia improves following RPS19 gene transfer"Molecular Therapy. 7・5. 613-622 (2003)

Hamaguchi I.、Flygare J. 等人：“核糖体蛋白 S19 (RPS19) 缺陷的 Diamond-Blackfan 贫血中多能造血祖细胞的增殖缺陷在 RPS19 基因转移后得到改善”7·5 (2003)。

Targeted disruption of the ribosomal protein S19 gene is lethal prior to implantation

• DOI: 10.1128/mcb.24.9.4032-4037.2004
• 发表时间: 2004-05-01
• 期刊: MOLECULAR AND CELLULAR BIOLOGY
• 影响因子: 5.3
• 作者: Matsson, H;Davey, EJ;Dahl, N
• 通讯作者: Dahl, N