Lentiviral vectors pseudotyped with a sindbis virus envelope glycoprotein

Abstract

Lentiviral vector particles comprising a Sindbis virus E2 glycoprotein variant and a lentiviral vector genome comprising a sequence of interest are provided. A lentiviral vector particle comprising: (a) an envelope comprising a Sindbis virus E2 glycoprotein variant; and (b) a lentiviral vector genome comprising a sequence of interest; wherein the E2 glycoprotein variant facilitates infection of dendritic cells by the lentiviral vector particle, and wherein the E2 glycoprotein variant has reduced binding to heparan sulfate compared to a reference sequence (HR strain).

Description

REFERENCE TO SEQUENCE LISTINGThe sequence listing of this patent application is provided separately in a file named “IDC203_SEQ_LISTING_ST25.txt”. The content of this file, which was created on 22 Jul. 2010 and consists of 150,110 bytes, is incorporated in its entirety.TECHNICAL FIELDThis patent application relates generally to targeted gene delivery, and more particularly to the use of a pseudotyped lentivirus comprising an envelope that targets dendritic cells and can thus be used for dendritic cell vaccination.BACKGROUNDDendritic cells (DCs) are essential antigen presenting cells for the initiation and control of immune responses. DCs can capture and process antigens, migrate from the periphery to a lymphoid organ, and present the antigens to resting T cells in a major histocompatibility complex (MHC)-restricted fashion. These cells are derived from bone marrow (BM) and display dendritic morphology and high mobility. The discovery of DCs as specialized antigen-presenting cells (A...

AI Technical Summary

Benefits of technology

This patent application is directed to pseudotyped lentiviral vectors that comprise a genome having a sequence of interest and an envelope comprising a glycoprotein of an arbovirus. The arbovirus glycoprotein can be from Sindbis virus, Dengue virus, and Venezuelan equine encephalitis virus. In particular, when the glycoprotein is an E2 protein from Sindbis virus, the E2 protein has at least one amino acid alteration at residue 160 compared to SEQ ID NO: 1. The amino acid alteration can be a deletion or an amino acid other than glutamic acid. The glycoprotein facilitates infection of dendritic cells. In all cases the E2 glycoprotein is not part of a fusion protein with Sindbis virus E3. In some embodiments, the E2 glycoprotein or variant binds DC-SIGN. The lentiviral vector also comprises a lentiviral genome that comprises a sequence of interest.

Problems solved by technology

All of these attempts however, involve the labor-intensive preparation of a patient-specific therapy that includes the loading of autologous DCs ex vivo with specific antigens, which are then administered to the patient.

There are a number of challenges however to achieving a safe and effective system.

The latter is a particular challenge in developing laboratory scale systems into products that can be produced by the pharmaceutical industry.

Though the SVGmu pseudotyped viral particles were able to selectively transduce cells expressing the DC-SIGN antigen, several aspects of the system make it unsuitable for therapeutic use.

For example, the E3 / E2 fusion protein displays an antigenic epitope of influenza hemagglutinin, and significantly, Sindbis virus strains with a mutation preventing correct processing of E3 from the E2 glycoprotein (so-called “pE2 mutants”), such as SVGmu, grow poorly in permissive cell lines and are severely attenuated in mouse pathogenicity.

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