Division of Gene Therapy, Research Center for Medical Sciences, The Jikei University School of Medicine, Japan; Department of Pediatrics, The Jikei University School of Medicine, Japan.
Division of Gene Therapy, Research Center for Medical Sciences, The Jikei University School of Medicine, Japan.
Mol Genet Metab. 2024 Jul;142(3):108494. doi: 10.1016/j.ymgme.2024.108494. Epub 2024 May 15.
Fabry disease (FD) is characterized by deficient activity of α-galactosidase A (GLA). Consequently, globotriaosylceramide (Gb3) accumulates in various organs, causing cardiac, renal, and cerebrovascular damage. Gene therapies for FD have been investigated in humans. Strong conditioning is required for hematopoietic stem cell-targeted gene therapy (HSC-GT). However, strong conditioning leads to various side effects and should be avoided. In this study, we tested antibody-based conditioning for HSC-GT in wild-type and FD model mice.
After preconditioning with an antibody-drug conjugate, HSC-GT using a lentiviral vector was performed in wild-type and Fabry model mice. In the wild-type experiment, the EGFP gene was introduced into HSCs and transplanted into preconditioned mice, and donor chimerism and EGFP expression were analyzed. In the FD mouse model, the GLA gene was introduced into HSCs and transplanted into preconditioned Fabry mice. GLA activity and Gb3 accumulation in the organs were analyzed.
In the wild-type mouse experiment, when anti-CD45 antibody-drug conjugate was used, the percentage of donor cells at 6 months was 64.5%, and 69.6% of engrafted donor peripheral blood expressed EGFP. When anti-CD117 antibody-drug conjugate and ATG were used, the percentage of donor cells at 6 months was 80.7%, and 73.4% of engrafted donor peripheral blood expressed EGFP. Although large variations in GLA activity among mice were observed in the FD mouse experiment for both preconditioning regimens, Gb3 was significantly reduced in many organs.
Antibody-based preconditioning may be an alternative preconditioning strategy for HSC-GT for treating FD.
法布里病（FD）的特征是α-半乳糖苷酶 A（GLA）活性缺乏。因此，糖鞘脂（Gb3）在各种器官中积累，导致心脏、肾脏和脑血管损伤。FD 的基因治疗已在人类中进行了研究。针对造血干细胞的基因治疗（HSC-GT）需要强烈的条件。然而，强烈的条件会导致各种副作用，应予以避免。在这项研究中，我们测试了针对野生型和 FD 模型小鼠的基于抗体的 HSC-GT 条件。
在抗体药物偶联物预处理后，使用慢病毒载体对野生型和 FD 模型小鼠进行 HSC-GT。在野生型实验中，将 EGFP 基因导入 HSCs 并移植到预处理的小鼠中，分析供体细胞嵌合和 EGFP 表达。在 FD 小鼠模型中，将 GLA 基因导入 HSCs 并移植到预处理的 FD 小鼠中。分析器官中的 GLA 活性和 Gb3 积累。
在野生型小鼠实验中，当使用抗 CD45 抗体药物偶联物时，6 个月时供体细胞的百分比为 64.5%，69.6%的移植供体外周血表达 EGFP。当使用抗 CD117 抗体药物偶联物和 ATG 时，6 个月时供体细胞的百分比为 80.7%，63.4%的移植供体外周血表达 EGFP。尽管在两种预处理方案的 FD 小鼠实验中，观察到 GLA 活性在小鼠之间存在很大差异，但许多器官中的 Gb3 明显减少。
基于抗体的预处理可能是治疗 FD 的 HSC-GT 的替代预处理策略。
