Department of Internal Medicine, Chungbuk National University School of Medicine, Cheongju, Korea.
Yonsei Med J. 2010 Jan;51(1):52-7. doi: 10.3349/ymj.2010.51.1.52. Epub 2009 Dec 29.
This study was designed to investigate whether transduction of lentiviral vectors (LV) carrying human coagulation factor VIII (hFVIII) cDNA into skeletal muscle could increase circulating hFVIII concentrations.
A LV containing bacterial LacZ gene as a control or human FVIII gene was intramuscularly administered into the thigh muscle of 5 weeks old Sparague-Dawley rats. The plasma human FVIII concentration and neutralizing anti-FVIII antibodies were measured for up to 12 weeks in these experimental animals.
The plasma human FVIII levels in the rats injected with LV carrying FVIII cDNA peaked at post-injection 1st week (5.19 +/- 0.14 ng/mL vs. 0.21 +/- 0.05 ng/mL in control rats , p < 0.05). Elevated hFVIII concentrations were maintained for 4 weeks (2.52 +/- 0.83 ng/mL vs. 0.17 +/- 0.08 ng/mL in control rats, p < 0.05) after a single intramuscular injection. In the Bethesda assay, neutralizing antibodies for FVIII protein were detected only in FVIII-LV injected rats by the 10th week, but not in control rats.
This study suggested that a single administration of an advanced generation LV carrying the human FVIII cDNA resulted in elevation of FVIII level in immune competent rats, and that this gene transfer approach to the skeletal muscle could be an effective tool in treatment of hemophilia A.
本研究旨在探讨将携带人凝血因子 VIII（hFVIII）cDNA 的慢病毒载体（LV）转导至骨骼肌是否能增加循环 hFVIII 浓度。
将含有细菌 LacZ 基因（作为对照）或人 FVIII 基因的 LV 通过肌肉内注射的方式注入 5 周龄的 SD 大鼠的股四头肌。在这些实验动物中，持续 12 周测量血浆人 FVIII 浓度和中和抗 FVIII 抗体。
注射携带 FVIII cDNA 的 LV 的大鼠血浆 hFVIII 水平在注射后第 1 周达到峰值（5.19±0.14ng/mL 比对照组的 0.21±0.05ng/mL，p<0.05）。单次肌肉内注射后，hFVIII 浓度可维持 4 周（2.52±0.83ng/mL 比对照组的 0.17±0.08ng/mL，p<0.05）。在 Bethesda 测定中，只有在第 10 周时才在接受 FVIII-LV 注射的大鼠中检测到针对 FVIII 蛋白的中和抗体，而在对照组大鼠中则未检测到。
本研究表明，单次给予携带人 FVIII cDNA 的先进代 LV 可导致免疫功能正常大鼠的 FVIII 水平升高，并且这种向骨骼肌的基因转移方法可能是治疗 A 型血友病的有效工具。
