Dataset Information

Assessment of F/HN-pseudotyped lentivirus as a clinically relevant vector for lung gene therapy.

ABSTRACT:

Rationale

Ongoing efforts to improve pulmonary gene transfer thereby enabling gene therapy for the treatment of lung diseases, such as cystic fibrosis (CF), has led to the assessment of a lentiviral vector (simian immunodeficiency virus [SIV]) pseudotyped with the Sendai virus envelope proteins F and HN.

Objectives

To place this vector onto a translational pathway to the clinic by addressing some key milestones that have to be achieved.

Methods

F/HN-SIV transduction efficiency, duration of expression, and toxicity were assessed in mice. In addition, F/HN-SIV was assessed in differentiated human air-liquid interface cultures, primary human nasal epithelial cells, and human and sheep lung slices.

Measurements and main results

A single dose produces lung expression for the lifetime of the mouse (~2 yr). Only brief contact time is needed to achieve transduction. Repeated daily administration leads to a dose-related increase in gene expression. Repeated monthly administration to mouse lower airways is feasible without loss of gene expression. There is no evidence of chronic toxicity during a 2-year study period. F/HN-SIV leads to persistent gene expression in human differentiated airway cultures and human lung slices and transduces freshly obtained primary human airway epithelial cells.

Conclusions

The data support F/HN-pseudotyped SIV as a promising vector for pulmonary gene therapy for several diseases including CF. We are now undertaking the necessary refinements to progress this vector into clinical trials.

SUBMITTER: Griesenbach U

PROVIDER: S-EPMC3530223 | biostudies-literature | 2012 Nov

REPOSITORIES: biostudies-literature

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Publications

Assessment of F/HN-pseudotyped lentivirus as a clinically relevant vector for lung gene therapy.

Griesenbach Uta U Inoue Makoto M Meng Cuixiang C Farley Raymond R Chan Mario M Newman Nikki K NK Brum Andrea A You Jun J Kerton Angela A Shoemark Amelia A Boyd A Christopher AC Davies Jane C JC Higgins Tracy E TE Gill Deborah R DR Hyde Stephen C SC Innes J Alastair JA Porteous David J DJ Hasegawa Mamoru M Alton Eric W F W EW

American journal of respiratory and critical care medicine 20120906 9

<h4>Rationale</h4>Ongoing efforts to improve pulmonary gene transfer thereby enabling gene therapy for the treatment of lung diseases, such as cystic fibrosis (CF), has led to the assessment of a lentiviral vector (simian immunodeficiency virus [SIV]) pseudotyped with the Sendai virus envelope proteins F and HN.<h4>Objectives</h4>To place this vector onto a translational pathway to the clinic by addressing some key milestones that have to be achieved.<h4>Methods</h4>F/HN-SIV transduction efficie ...[more]

PMID: 22955314

Similar Datasets

Project description:BackgroundHendra virus (HeV) and Nipah virus (NiV) are newly emerged zoonotic paramyxoviruses discovered during outbreaks in Queensland, Australia in 1994 and peninsular Malaysia in 1998/9 respectively and classified within the new Henipavirus genus. Both viruses can infect a broad range of mammalian species causing severe and often-lethal disease in humans and animals, and repeated outbreaks continue to occur. Extensive laboratory studies on the host cell infection stage of HeV and NiV and the roles of their envelope glycoproteins have been hampered by their highly pathogenic nature and restriction to biosafety level-4 (BSL-4) containment. To circumvent this problem, we have developed a henipavirus envelope glycoprotein pseudotyped lentivirus assay system using either a luciferase gene or green fluorescent protein (GFP) gene encoding human immunodeficiency virus type-1 (HIV-1) genome in conjunction with the HeV and NiV fusion (F) and attachment (G) glycoproteins.ResultsFunctional retrovirus particles pseudotyped with henipavirus F and G glycoproteins displayed proper target cell tropism and entry and infection was dependent on the presence of the HeV and NiV receptors ephrinB2 or B3 on target cells. The functional specificity of the assay was confirmed by the lack of reporter-gene signals when particles bearing either only the F or only G glycoprotein were prepared and assayed. Virus entry could be specifically blocked when infection was carried out in the presence of a fusion inhibiting C-terminal heptad (HR-2) peptide, a well-characterized, cross-reactive, neutralizing human mAb specific for the henipavirus G glycoprotein, and soluble ephrinB2 and B3 receptors. In addition, the utility of the assay was also demonstrated by an examination of the influence of the cytoplasmic tail of F in its fusion activity and incorporation into pseudotyped virus particles by generating and testing a panel of truncation mutants of NiV and HeV F.ConclusionsTogether, these results demonstrate that a specific henipavirus entry assay has been developed using NiV or HeV F and G glycoprotein pseudotyped reporter-gene encoding retrovirus particles. This assay can be conducted safely under BSL-2 conditions and will be a useful tool for measuring henipavirus entry and studying F and G glycoprotein function in the context of virus entry, as well as in assaying and characterizing neutralizing antibodies and virus entry inhibitors.

Dataset Information

Assessment of F/HN-pseudotyped lentivirus as a clinically relevant vector for lung gene therapy.

Rationale

Objectives

Methods

Measurements and main results

Conclusions

Publications

Assessment of F/HN-pseudotyped lentivirus as a clinically relevant vector for lung gene therapy.

Similar Datasets

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