Abstract
We have shown that a lentiviral vector (rSIV.F/HN) pseudotyped with the F and HN proteins from Sendai virus generates high levels of intracellular proteins after lung transduction. Here, we evaluate the use of rSIV.F/HN for production of secreted proteins. We assessed whether rSIV.F/HN transduction of the lung generates therapeutically relevant levels of secreted proteins in the lung and systemic circulation using human α1-anti-trypsin (hAAT) and factor VIII (hFVIII) as exemplars. Sedated mice were transduced with rSIV.F/HN carrying either the secreted reporter gene Gaussia luciferase or the hAAT or hFVIII cDNAs by nasal sniffing. rSIV.F/HN-hAAT transduction lead to therapeutically relevant hAAT levels (70 μg/ml) in epithelial lining fluid, with stable expression persisting for at least 19 months from a single application. Secreted proteins produced in the lung were released into the circulation and stable expression was detectable in blood. The levels of hFVIII in murine blood approached therapeutically relevant targets. rSIV.F/HN was also able to produce secreted hAAT and hFVIII in transduced human primary airway cells. rSIV.F/HN transduction of the murine lungs leads to long-lasting and therapeutically relevant levels of secreted proteins in the lung and systemic circulation. These data broaden the use of this vector platform for a large range of disease indications.
Highlights
Our ongoing efforts to improve pulmonary gene transfer for the treatment of lung diseases such as cystic fibrosisThese authors contributed : Michael C
We have previously shown that the cationic lipid formulation GL67A stabilises lung disease in CF patients [20], and leads to expression of the secreted reporter gene Gaussia luciferase (GLux) in murine and ovine lungs [21]
Plasmids that are depleted of CpGs (CG dinucleotides) reduce inflammation and achieve longer-lasting gene expression compared with CpG-containing plasmids in mouse lung in vivo [22] and we removed all CpGs from the human alpha-1 antitrypsin (hAAT) complementary DNA (cDNA)-generating sohAAT
Summary
Our ongoing efforts to improve pulmonary gene transfer for the treatment of lung diseases such as cystic fibrosisThese authors contributed : Michael C. Our ongoing efforts to improve pulmonary gene transfer for the treatment of lung diseases such as cystic fibrosis. (DNAVEC Centre), Tsukuba, Japan (CF), have led to the development of a novel lentiviral vector (simian immunodeficiency virus [SIV]) pseudotyped with the Sendai virus (SeV) envelope proteins F and HN (recombinant (r)SIV.F/HN). This vector can drive high, stable levels of reporter gene expression in the lungs and nose of mice for the duration of their lifetime (~ 2 yr) and, in contrast to other viral vectors, repeated administration does not lead to loss of efficacy [1,2,3]. The efficacy and toxicity profile of the vector, supports progression toward the clinic
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