n .One of the key issues for treatment of central nervous system (CMS)diseases is finding effective means to deliver drugs through the blood-brain barrier (BBB). This block is even more difficult to circumvent when delivering large macromolecules such as neurotrophic factors, nucleic acid based therapeutic reagentsor gene therapy vectors that encode such factors. The first step towards effective delivery of therapeutic genes to the CMSwould be the development of gene therapy vectors that target the CMS specifically. Ideally, these vectors could be injected directly into the bloodstream and home to the blood-brain barrier, transduce the capillary endothelial cells and either express their therapeutic genes within the endothelial cells or following delivery into the brain. Towards this end, we made significant progress in developing lentiviral targeting vectors for stable introduction of genes into specified cells and tissues. The central hypothesis of this proposal is that efficient targeting vectors with high selectivity for the CNS can be developed to deliver therapeutic genes into the CNS. We recently reported that a lentiviral vector pseudotypedwith a modified Sindbis virus envelope bearing the Fc binding domain of protein A can be retargeted based upon the specificity of monoclonal antibodies directed to specific cell surface molecules. As proof of principle, we demonstrated that this vector can be injected intravenously into the mouse tail vein and specifically transduced metastatic melanoma cells bearing P-glycoprotein as the target molecule (Morizono K, et. al,. 2005 Lentiviral vector retargeting to P- Glycoprotein on metastatic melanoma via intravenous injection. Nat Med. 11(3):346-52.). Because the approach can be generalized to other ligand/antibody combinations we propose here to develop lentiviral gene therapy vectors that will specifically target the blood-brain barrier. In that regard, our preliminary studies indicate that further modification of the targeting vector and conjugation with anti-transferrin receptor (TfR) antibodies can result in targeting through the bloodstream to what appears to be specifically the CNS. Given our long-standing interest in retroviral biology, recently applied to gene therapy vectors, and the novel targeting approach described here, we feel that we are well positioned to address the questions posed in this application.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21NS055212-02
Application #
7337084
Study Section
Gene and Drug Delivery Systems Study Section (GDD)
Program Officer
Jacobs, Tom P
Project Start
2007-01-01
Project End
2008-12-31
Budget Start
2008-01-01
Budget End
2008-12-31
Support Year
2
Fiscal Year
2008
Total Cost
$202,781
Indirect Cost
Name
University of California Los Angeles
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
092530369
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Morizono, Kouki; Chen, Irvin S Y (2011) Receptors and tropisms of envelope viruses. Curr Opin Virol 1:13-8
Morizono, Kouki; Xie, Yiming; Olafsen, Tove et al. (2011) The soluble serum protein Gas6 bridges virion envelope phosphatidylserine to the TAM receptor tyrosine kinase Axl to mediate viral entry. Cell Host Microbe 9:286-98
Morizono, Kouki; Ku, Amy; Xie, Yiming et al. (2010) Redirecting lentiviral vectors pseudotyped with Sindbis virus-derived envelope proteins to DC-SIGN by modification of N-linked glycans of envelope proteins. J Virol 84:6923-34
Morizono, Kouki; Xie, Yiming; Helguera, Gustavo et al. (2009) A versatile targeting system with lentiviral vectors bearing the biotin-adaptor peptide. J Gene Med 11:655-63
Morizono, Kouki; Pariente, Nonia; Xie, Yiming et al. (2009) Redirecting lentiviral vectors by insertion of integrin-tageting peptides into envelope proteins. J Gene Med 11:549-58