Establishment of a basis for therapeutic treatment of chronic granulomatous disease using human induced pluripotent stem cells

建立利用人诱导多能干细胞治疗慢性肉芽肿病的基础

• 批准号: 24791102
• 负责人: INAKI Makoto
• 金额: $ 2.75万
• 依托单位: National Research Institute for Child Health and Development
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2012
• 资助国家: 日本
• 起止时间: 2012-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-24791102/
• 关键词: iPS細胞; 疾患モデリング; 慢性肉芽腫症

We tried to model chronic granulomatous disease using disease-specific induced pluripotent stem cells (iPS cells). At first, we generated patient specific iPS cells from T cells in 4 patients' peripheral blood and CYBB gene knockout iPS cells by using genome editing technology. The CYBB knockout iPS cells could be differentiated into neutrophil that cannot generate reactive oxygen species. However, macrophage or dendritic cells differentiated from CYBB gene knockout iPS cells in this study did not exhibit any significant abnormality. With consideration for the possibility that the origin of iPS cells can affect results of disease modeling, a system for generating iPS cells from various blood cells was established. Finally, a system for homologous recombination in iPS cells using adeno-associated virus was also established. These experimental systems could be a platform for a therapeutic treatment of chronic granulomatous disease.

我们试图使用疾病特异性诱导多能干细胞(iPS细胞)来建立慢性肉芽肿性疾病的模型。首先，我们利用基因组编辑技术，从4例患者外周血中的T细胞和CYBB基因敲除的iPS细胞中获得了患者特异性的iPS细胞。基因敲除的iPS细胞可以分化为不能产生活性氧的中性粒细胞。然而，本研究中从CYBB基因敲除的iPS细胞分化出的巨噬细胞或树突状细胞没有表现出任何明显的异常。考虑到iPS细胞的来源可能会影响疾病模型的结果，建立了一种从不同血细胞生成iPS细胞的系统。最后，还建立了用腺相关病毒在iPS细胞中进行同源重组的系统。这些实验系统可能成为治疗慢性肉芽肿性疾病的平台。

项目成果

Primary human B lymphocyte-derived iPS cells, their generation and application

原代人B淋巴细胞来源的iPS细胞、其产生和应用

• 发表时间: 2013
• 作者: Makoto Inaki;Masafumi Onodera;Makoto Inaki
• 通讯作者: Makoto Inaki

GENERATION OF INDUCED PLURIPOTENT STEM CELLS FROM PRIMARY HUMAN B LYMPHOCYTES

从原代人 B 淋巴细胞诱导产生多能干细胞

• 发表时间: 2013
• 作者: Makoto Inaki;Masafumi Onodera
• 通讯作者: Masafumi Onodera

Gene transfer to human hematopoietic cells with GaLV pseudotyped retroviruses concentrated from serum-free supernatants by PEG precipitation with polyvalent immunoglobulin

使用多价免疫球蛋白 PEG 沉淀从无血清上清液中浓缩的 GaLV 假型逆转录病毒将基因转移至人造血细胞

• 发表时间: 2012
• 作者: Makoto Inaki;Masafumi Onodera
• 通讯作者: Masafumi Onodera

Gene transfer to human hematopoietic cells with GaLV pseudotyped retroviruses concentrated from serum-free supernatants by PEG precipitation with polyvalent immunoglobulin.

使用 GaLV 假型逆转录病毒将基因转移至人类造血细胞，该病毒通过多价免疫球蛋白 PEG 沉淀从无血清上清液中浓缩。

• 发表时间: 2012
• 作者: Makoto Inaki;Masafumi Onodera
• 通讯作者: Masafumi Onodera