Gene therapy for chronic granulomatous disease combined with in vivo expansion of transduced hematopoietic cells.

慢性肉芽肿性疾病的基因治疗结合转导造血细胞的体内扩增。

• 批准号: 14570768
• 负责人: KUME Akihiro
• 金额: $ 2.24万
• 依托单位: Jichi Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570768/
• 关键词: gene therapy; chronic granulomatous disease; translational research; hematopoietic stem cells; selecvtive amplifier gene; retroviral vector; innate immunity; phagocytes; 活性酸素; 移植・再生医療

We have developed 'selective amplifier genes (SAGs)' which encode chimeric receptors to confer in vivo growth advantage on transduced hematopoietic cells. In this project, we investigated the feasibility of using SAGs to boost clinical benefit of gene therapy in a mouse model of X-linked chronic granulomatous disease (X-CGD). (1)A fusion receptor GcRER was constructed with the granulocyte colony-stimulating factor receptor and the estrogen-binding domain. X-CGD bone marrow cells were transduced by a retroviral vector encoding GcRER and gp91-phox, the latter of which is deficient in X-CGD. The transduced cells were reinfused into irradiated X-CGD mice for hematopoietic reconstitution. After recovery, estrogen was administered to a subset of the transplants, and the estrogen-treated animals had a significantly higher level of functionally corrected granulocytes. (2)As a second generation of SAGs, the extracellular domain of erythropoietin receptor (EpoR) was employed as a molecular switch to regulate cell growth signaling. EpoR was fused to the cytoplasmic domain of c-Mpl, and the gene for the chimera (EpoRMpl) was inserted into a retroviral vector together with the gp91-phox gene. X-CGD bone marrow cells were transduced with the vector and transplanted to X-CGD recipients as in the previous experiments. Erythropoietin administration to the recipient animals resulted in an elevation of functionally corrected granulocytes.These results indicated that SAG-mediated expansion of transduced hematopoietic cells is feasible in gene therapy for X-CGD.

我们已经开发了“选择性放大基因（SAG）”，其编码嵌合受体，以赋予转导的造血细胞体内生长优势。在这个项目中，我们研究了在X连锁慢性肉芽肿病（X-CGD）小鼠模型中使用SAGs来提高基因治疗临床效益的可行性。(1)A用粒细胞集落刺激因子受体和雌激素结合结构域构建融合受体GcRER。用编码GcRER和gp 91-phox的逆转录病毒载体转导X-CGD骨髓细胞，后者在X-CGD中是缺陷的。将转导的细胞回输到经照射的X-CGD小鼠中用于造血重建。恢复后，雌激素被给予一个子集的移植，和雌激素治疗的动物有一个显着更高的水平的功能校正粒细胞。(2)As第二代SAG，促红细胞生成素受体的胞外结构域（EpoR）被用作调节细胞生长信号的分子开关。将EpoR与c-Mpl的胞质结构域融合，并将嵌合体的基因（EpoRMpl）与gp 91-phox基因一起插入逆转录病毒载体中。用载体转导X-CGD骨髓细胞，并如先前实验中那样移植到X-CGD受体。向受体动物给予促红细胞生成素导致功能纠正的粒细胞增加。这些结果表明，SAG介导的转导造血细胞扩增在X-CGD的基因治疗中是可行的。

项目成果

Hara T, et al.: "Expansion of genetically corrected neutrophils in chronic granulomatous disease mice by cotransferring a therapeutic gene and a selective amplifier gene"Gene Therapy. in press. (2004)

Hara T 等人：“通过共转移治疗基因和选择性扩增基因，在慢性肉芽肿病小鼠中扩增基因校正的中性粒细胞”基因疗法。

Mochizuki S, et al.: "Long-term correction of hiperphenylalaninemia by AAV-mediated gene transfer leads to behavioral recovery in phenylketonuria mice"Gene Therapy. in press. (2004)

Mochizuki S 等人：“通过 AAV 介导的基因转移长期纠正高苯丙氨酸血症可导致苯丙酮尿症小鼠的行为恢复”基因治疗。

Kume A, et al.: "In vivo expansion of transduced murine hematopoietic cells with a selective amplifier gene."The Journal of Gene Medicine. 5. 175-181 (2003)

Kume A 等人：“用选择性放大器基因对转导的小鼠造血细胞进行体内扩增。”《基因医学杂志》。

Nagashima T et al.: "New selective amplifier genes containing c-mpl for hematopoietic cell expansion."Biochem Biophys Res Commun. 303(1). 170-176 (2003)

Nagashima T 等人：“含有用于造血细胞扩增的 c-mpl 的新型选择性扩增基因。”Biochem Biophys Res Commun。

Hara T et al.: "Expansion of genetically corrected neutrophils in chronic granulomatous disease mice by cotransferring herapeutic gene and a selective amplifier gene."Gene Ther. (in press).

Hara T 等人：“通过共转移治疗基因和选择性扩增基因，在慢性肉芽肿病小鼠中扩增基因校正的中性粒细胞。”Gene Ther。