Preclinical protocol for in vivo selection of hematopoietic stem cells corrected by gene therapy in Fanconi anemia group C
Preclinical protocol for in vivo selection of hematopoietic stem cells corrected by gene therapy in Fanconi anemia group C
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DOI：
10.1006/mthe.2000.0226
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发表时间：
2001-01-01
影响因子：
12.4
通讯作者：
Grompe, M
中科院分区：
文献类型：
--
作者：
Noll, M;Bateman, RL;Grompe, M
范可尼贫血 (FA) 是一种常染色体隐性遗传疾病，其特征是出生缺陷、恶性肿瘤发病率增加、进行性骨髓衰竭和细胞对 DNA 交联剂过敏。骨髓移植具有治疗作用，因此 FA 是造血基因治疗的候选疾病。我们之前使用丝裂霉素C（MMC）实现了移植到FANCC敲除小鼠体内的野生型造血干细胞（HSC）的体内选择。然而，MMC 在人类 FA 基因治疗中的临床应用不太可能，因为其对人类 FA 患者的毒性特征未知。相比之下，环磷酰胺 (CPA) 和伽马射线照射 (IR) 已用于人类 FA 患者，因此我们测试了这些方案在体内实现基因校正 HSC 的选择的能力。我们发现非清髓剂量的 CPA 或 IR 或 CPA + IR 的组合对于实现移植野生型 HSC 的体内选择非常有效。此外，这种无毒方案还选择了通过离体逆转录病毒基因治疗纠正的FANCC突变HSC。我们建议这些无毒剂量的 CPA 和/或 IR 也可用于增强人类 FA 患者的基因治疗。
Fanconi anemia (FA) is an autosomal recessive disorder characterized by birth defects, Increased incidence of malignancy, progressive bone marrow failure, and cellular hypersensitivity to DNA cross-linking agents. Bone marrow transplantation is therapeutic and therefore FA is a candidate disease for hematopoietic gene therapy. We have previously used mitomycin C (MMC) to achieve in vivo selection of wild-type hematopoietic stem cells (HSC) transplanted into FANCC knockout mice. However, clinical application of MMC in human FA gene therapy is unlikely because of its unknown toxicity profile in human FA patients. In contrast, cyclophosphamide (CPA) and gamma -irradiation (IR) are already in use with human FA patients and we therefore tested these regimens for their ability to achieve selection of genetically corrected HSCs in vivo. We found that nonmyeloablative doses of CPA or IR or combinations of CPA + IR were highly efficient at achieving in vivo selection of transplanted wild-type HSC. Furthermore, this nontoxic regimen also selected FANCC-mutant HSC corrected by ex vivo retroviral gene therapy. We suggest those nontoxic doses of CPA and/or IR could also be used to enhance gene therapy in human FA patients.