Delivery of Factor VIII Gene into Skeletal Muscle Cells Using Lentiviral Vector

Abstract

PurposeThis 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.Materials and MethodsA 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.ResultsThe 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.ConclusionThis 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.

A novel gene therapy strategy using secreted multifunctional anti-HIV proteins to confer protection to gene-modified and unmodified target cells

“…evaluated for the treatment of other diseases, such as erythropoietin deficiency, a-1 antitrypsin deficiency, hemophilia A and Duchenne muscular dystrophy. [73][74][75][76] Skin and muscle tissues are readily accessible and are highly vascularized, making them suitable for systemic delivery of therapeutic proteins via circulation. Indeed, serum levels of therapeutic proteins reached 1000 mg ml À 1 in vivo.…”

Induced Pluripotent Stem Cells: Therapeutic Applications in Monogenic and Metabolic Diseases, and Regulatory and Bioethical Considerations

“…In fact, haemophilia is now recognized as a condition amenable to gene therapy [61][62][63][64]. Strategies available include use of lentiviral (LVV) [65] and adeno-associated (AAV) [66] vectors in adult stem cells and autologous fibroblasts, in platelets and in hematopoietic stem cells; transfer by means of non-viral vectors; and repair of mutations with chimeric oligonucleotides. The studies published so far have, in the most part, not reported any severe adverse effect resulting from the application of such strategies in the clinical trials performed.…”

Advanced therapies for the treatment of hemophilia: future perspectives

“…Since those results were published, use of lentiviral vectors has not ceased to grow. Thus, Jeon et al [36] transduced this type of viral vector into skeletal muscle to increase factor VIII expression. Factor VIII plasma levels at one week post-injection were 5.19 ng/mL vs 0.21 ng/mL in control rats, with those levels staying constant over 4 weeks with a single dose of the vector.…”