D'Amico Andrea, Blits Bas, van den Herik Joëlle, de Winter Fred, Kaijen Paul H P, Hobo Barbara, van Asbeck Alexander Henrik, Verhaagen Joost, Mosmuller David G M, Voorberg Jan
Department of Molecular Hematology, Sanquin Research and Landsteiner Laboratory of the Amsterdam UMC, 1066 CX Amsterdam, the Netherlands.
SanaGen B.V., 1066 CX Amsterdam, the Netherlands.
Mol Ther Methods Clin Dev. 2025 Jun 25;33(3):101523. doi: 10.1016/j.omtm.2025.101523. eCollection 2025 Sep 11.
Factor VII (FVII) deficiency is a rare bleeding disorder with a prevalence of approximately 1:300,000-500,000 individuals. We explored whether adeno-associated virus (AAV)-mediated gene therapy can achieve durable and functional expression of human FVII (hFVII) . Wild-type hFVII (hFVIIwt) and a naturally occurring splice variant designated hFVII(-22) (GenBank: NM_019616.4) were expressed under the control of the Apolipoprotein E-derived hepatic locus control region and the α1-anti-trypsin promoter. Expression cassettes were packaged in either recombinant AAV5 (rAAV5) or recombinant AAV8 (rAAV8). For both hFVIIwt and hFVII(-22), 5- to 20-fold higher plasma levels of hFVII could be obtained when rAAV8 was used as a vector as opposed to rAAV5. Interestingly, hFVII levels obtained by employing rAAV8 expressing the hFVII(-22) cDNA variant were approximately 10 times higher than those obtained using rAAV8 expressing hFVIIwt. Based on these results, we generated an rAAV8-based gene therapy vector encoding hFVII(-22) and evaluated long-term expression . Employing a vector dose of 0.8 × 10 genome copies (gc)/kg, we observed 48 weeks of functional hFVII expression which peaked at 16 IU/mL and stabilized at 7 IU/mL. These results support the pre-clinical development of AAV8-mediated delivery employing the splice-variant hFVII(-22) for patients suffering from FVII deficiency.
凝血因子VII（FVII）缺乏症是一种罕见的出血性疾病，患病率约为1：300,000 - 500,000人。我们探讨了腺相关病毒（AAV）介导的基因治疗是否能够实现人FVII（hFVII）的持久和功能性表达。野生型hFVII（hFVIIwt）和一种天然存在的剪接变体hFVII(-22)（GenBank：NM_019616.4）在载脂蛋白E衍生的肝脏位点控制区和α1-抗胰蛋白酶启动子的控制下表达。表达盒被包装在重组AAV5（rAAV5）或重组AAV8（rAAV8）中。对于hFVIIwt和hFVII(-22)，与rAAV5相比，当使用rAAV8作为载体时，血浆中hFVII水平可提高5至20倍。有趣的是，使用表达hFVII(-22) cDNA变体的rAAV8获得的hFVII水平比使用表达hFVIIwt的rAAV8获得的水平高约10倍。基于这些结果，我们构建了一种基于rAAV8的编码hFVII(-22)的基因治疗载体并评估了其长期表达。使用0.8×10基因组拷贝（gc）/kg的载体剂量，我们观察到功能性hFVII表达持续了48周，峰值为16 IU/mL，并稳定在7 IU/mL。这些结果支持了AAV8介导的采用剪接变体hFVII(-22)对FVII缺乏症患者进行递送的临床前开发。
