Center for Medical Genetics, School of Life Sciences, Central South University, Changsha 410078, China.
Int J Mol Sci. 2024 Mar 24;25(7):3635. doi: 10.3390/ijms25073635.
Hemophilia A (HA) is a common X-linked recessive hereditary bleeding disorder. Coagulation factor VIII (FVIII) is insufficient in patients with HA due to the mutations in the gene. The restoration of plasma levels of FVIII via both recombinant B-domain-deleted FVIII (BDD-FVIII) and B-domain-deleted () transgenes was proven to be helpful. FVIII-Padua is a 23.4 kb tandem repeat mutation in the associated with a high gene expression and thrombogenesis. Here we screened a core enhancer element in FVIII-Padua for improving the expression. In detail, we identified a 400 bp efficient enhancer element, C400, in FVIII-Padua for the first time. The core enhancer C400 extensively improved the transcription of driven by human elongation factor-1 alpha in HepG2, HeLa, HEK-293T and induced pluripotent stem cells (iPSCs) with different genetic backgrounds, as well as iPSCs-derived endothelial progenitor cells (iEPCs) and iPSCs-derived mesenchymal stem cells (iMSCs). The expression of FVIII protein was increased by C400, especially in iEPCs. Our research provides a novel molecular target to enhance expression of FVIII protein, which has scientific value and application prospects in both viral and nonviral HA gene therapy strategies.
血友病 A（HA）是一种常见的 X 连锁隐性遗传性出血性疾病。由于 基因的突变，HA 患者的凝血因子 VIII（FVIII）不足。通过重组 B 域缺失的 FVIII（BDD-FVIII）和 B 域缺失的 （）转基因，可恢复 FVIII 的血浆水平，这已被证明是有效的。FVIII-Padua 是一种与高 基因表达和血栓形成相关的 23.4 kb 串联重复突变。在这里，我们筛选了 FVIII-Padua 中的核心增强子元件，以提高 表达。具体来说，我们首次在 FVIII-Padua 中鉴定了一个 400 bp 的有效增强子元件 C400。核心增强子 C400 广泛提高了人延伸因子-1 ɑ 在 HepG2、HeLa、HEK-293T 和具有不同遗传背景的诱导多能干细胞（iPSCs）以及 iPSCs 衍生的内皮祖细胞（iEPCs）和 iPSCs 衍生的间充质干细胞（iMSCs）中驱动的 转录，C400 还增加了 FVIII 蛋白的表达，尤其是在 iEPCs 中。我们的研究为增强 FVIII 蛋白的表达提供了一个新的分子靶点，在病毒和非病毒 HA 基因治疗策略中具有科学价值和应用前景。
