Yuan Haokun, Wu Xiaomei, Liu Hongwei, Chang Lung-Ji
School of Medicine, University of Electronic Science and Technology of China, Chengdu, China.
Shenzhen Geno-Immune Medical Institute, Shenzhen, China.
Hum Gene Ther. 2023 Jan;34(1-2):19-29. doi: 10.1089/hum.2022.140. Epub 2022 Dec 27.
Chronic granulomatous disease (CGD) is a rare congenital immunodeficiency characterized by a defect in nicotinamide adenine dinucleotide phosphate oxidase required for phagocytosis. Hematopoietic stem cell (HSC) transplantation is currently the only curative treatment, but it is ladened with morbidities and mortality. Gene therapy is a promising treatment for CGD. However, if not properly designed, the gene therapy approach may not be successful. We engineered lentiviral vectors (LVs) carrying a universal promoter () and two myeloid-specific promoters ( and ) to drive the expression of green fluorescence protein (GFP) or , one of the key defective genes causing CGD. Tissue-specific LV expression was investigated and in a CGD mouse model. We compared GFP expression in both myeloid differentiated and undifferentiated HSCs. The CGD mice were transplanted with LV-modified mouse HSCs to investigate expression of and restoration of reactive oxygen species. The LV promoters were further compared under low and high-transgenic conditions to assess safety and therapeutic efficacy. A pneumonia disease model based on pathogenic challenge was established to assess the survival rate and body weight change. All three promoters demonstrated ectopic expression and . The promoter showed the highest expression of GFP or in transduced cells, including HSCs without cytotoxicity, whereas the LV- showed the highest transgene delivery efficiency with high myeloid specificity. Importantly, under low-transgenic condition, only the LV showed high antibacterial activity .
慢性肉芽肿病（CGD）是一种罕见的先天性免疫缺陷病，其特征是吞噬作用所需的烟酰胺腺嘌呤二核苷酸磷酸氧化酶存在缺陷。造血干细胞（HSC）移植是目前唯一的治愈性治疗方法，但存在发病率和死亡率。基因治疗是治疗CGD的一种有前景的方法。然而，如果设计不当，基因治疗方法可能不会成功。我们构建了携带通用启动子（）和两个髓系特异性启动子（和）的慢病毒载体（LV），以驱动绿色荧光蛋白（GFP）或导致CGD的关键缺陷基因之一的表达。在CGD小鼠模型中研究了组织特异性LV表达。我们比较了髓系分化和未分化HSC中GFP的表达。将LV修饰的小鼠HSC移植到CGD小鼠中，以研究的表达和活性氧的恢复。在低转基因和高转基因条件下进一步比较LV启动子，以评估安全性和治疗效果。建立了基于致病性攻击的肺炎疾病模型，以评估存活率和体重变化。所有三种启动子均表现出异位表达和。启动子在转导细胞（包括无细胞毒性的HSC）中显示出最高的GFP或表达，而LV-显示出最高的转基因递送效率和高髓系特异性。重要的是，在低转基因条件下，只有LV显示出高抗菌活性。
