Vectors and methods for gene transfer

Abstract

The present invention provides a recombinant adenovirus comprising coat proteins that lack native binding. In particular, the present invention provides a recombinant adenovirus comprising a penton base protein and a fiber protein, wherein the penton base protein and the fiber protein lack native binding. The present invention further provides a recombinant adenovirus comprising (a) a penton base protein that lacks native binding and (b) a nonnative amino acid sequence that binds a cell-surface binding site.

Description

CROSS REFERENCE TO RELATED PATENT APPLICATIONS [0001] The present application is a continuation of co-pending U.S. patent application Ser. No. 09 / 999,724, filed Oct. 24, 2001, which, in turn, is a continuation of U.S. patent application Ser. No. 09 / 101,751, filed Jan. 29, 1999, now U.S. Pat. No. 6,465,253, which, in turn, is the national phase of International Patent Application PCT / US96 / 19150, designating the U.S. and filed Nov. 27, 1996; a continuation-in-part of U.S. patent application Ser. No. 08 / 563,368, filed Nov. 28, 1995, now U.S. Pat. No. 5,965,541; a continuation-in-part of U.S. patent application Ser. No. 08 / 701,124, filed Aug. 21, 1996, now U.S. Pat. No. 5,846,782; and a continuation-in-part of U.S. patent application Ser. No. 08 / 700,846, filed Aug. 21, 1996, now U.S. Pat. No. 5,962,311, which, in turn, is a continuation-in-part of U.S. patent application Ser. No. 08 / 634,060, filed Apr. 17, 1996, now U.S. Pat. No. 5,712,136, which, in turn, is a continuation-in-part of U...

AI Technical Summary

Benefits of technology

[0015] The present invention provides a chimeric adenoviral coat protein (e.g., a fiber, hexon or penton protein), which differs from the wild-type (i.e., native) fiber protein by the introduction of a nonnative amino acid sequence, preferably at or near the carboxyl terminus. The resultant chimeric adenovirus coat protein is able to direct entry into cells of a vector comprising the coat protein that is more efficient than entry into cells of a vector that is identical except for comprising a wild-type adenovirus coat protein rather than the chimeric adenovirus coat protein. One direct result of this increased efficiency of entry is that the chimeric adenovirus coat protein enables the adenovirus to bind to and enter numerous cell types which adenovirus comprising wild-type coat protein typically cannot enter or can enter with only a low efficiency. The present invention also provides an adenoviral vector that comprises the chimeric adenovirus coat protein, and methods of constructing and using such a vector.

Problems solved by technology

A drawback to adenovirus-mediated gene therapy is that significant decreases in gene expression are observed after two weeks following administration of the vector.

This antibody response against the virus can prevent effective re-administration of the viral vector.

Another drawback of using recombinant adenovirus in gene therapy is that certain cells are not readily amenable to adenovirus-mediated gene delivery.

This lack of ability to infect all cells has lead researchers to seek out ways to introduce adenovirus into cells that cannot be infected by adenovirus, e.g. due to lack of adenoviral receptors.

However, these approaches are disadvantageous in that they require additional steps that covalently link large molecules, such as polylysine, receptor ligands, and antibodies, to the virus (Cotten (1992), supra; Wagner et al., Proc. Natl. Acad. Sci., 89, 6099-6103 (1992)).

This adds to the size of the resultant vector as well as its cost of production.

Moreover, the targeted particle complexes are not homogeneous in structure, and their efficiency is sensitive to the relative ratios of viral particles, linking molecules, and targeting molecules used.

Thus, this approach for expanding the repertoire of cells amenable to adenoviral-mediated gene therapy is less than optimal.

However, no studies have confirmed this point to date.

In fact, studies have suggested that adenovirus gene delivery to differentiated lung epithelium is less efficient than delivery to proliferating or to undifferentiated cells (Grubb et al., supra; Dupuit et al., supra).

The requirement for a high MOI to achieve transduction is disadvantageous inasmuch as any immune response associated with adenoviral infection necessarily would be exacerbated with use of high doses.

Images