Title:  Genetic suppressor elements against human immunodeficiency virus

United States Patent:  6,316,210

Assignee:  Subsidiary No. 3, Inc. (Wilminton, NC)

Filed:  September 1, 1999

The present invention relates to genetic elements that suppress the activities of the human immunodeficiency virus (HIV). In particular, the invention relates to polynucleotides isolated from the HIV-1 genome, methods for isolating, identifying and designing such polynucleotides, and methods for using them for the protection of human cells against HIV infection and/or replication. The present invention also relates to polynucleotides that prevent tumor cell formation and the use of such polynucleotides to prevent tumorigenesis.

SUMMARY OF THE INVENTION

The present invention relates to specific HIV-derived polynucleotides herein referred to as GSEs that suppress HIV infection and/or replication in human cells, methods for isolating and identifying such polynucleotides, methods for designing such polynucleotides, and methods for using them in the prevention or treatment of HIV infection.

The invention is based, in part, on the Applicants' discovery that nucleotide fragments can be isolated from the HIV-1 genome, based on their ability to suppress the activation of latent HIV-1 in a CD4⁺ cell line or on their ability to prevent cells from productive infection by HIV-1. In this connection, any cellular or viral marker associated with HIV replication can be used to monitor the activation of latent HIV or productive infection of cells. A number of novel HIV-1 GSE polynucleotides are selected on the basis of their ability to sustain CD4 expression by the induced cells, and several of such sequences are further shown to protect uninfected T cells from productive infection by HIV-1. The GSEs may function in the form of an RNA product or protein product, both of which are within the scope of the invention. Another exemplary marker is the intracellular p24. Replication of HIV in susceptible cells is associated with accumulation of intracellular p24, concomitant with down modulation of surface CD4. The expression of GSEs capable of interfering with productive infection should result in enrichment of protected cells displaying the CD4 +, p24^- phenotype. Such cells can be separated by FACS from infected population.

This invention is also based upon the discovery that isolated GSEs cluster around narrowly defined regions of the HIV-1 genome, particularly, the surprising discovery that inhibitory or protective clusters of GSEs were also isolated from previously untargeted regions (RT and nef). Preferred embodiments of the invention provide methods for designing GSEs based upon the consensus sequences of such GSE clusters.

A wide range of uses are encompassed by the invention, including but not limited to, AIDS treatment and prevention by transferring GSE into HIV-1-susceptible cell types. For example, GSE may be transferred into T cells or hematopoietic progenitor cells in vitro followed by their engraftment in an autologous, histocompatible or even histoincompatibile recipient. In an alternative embodiment of the invention, any cells susceptible to HIV infection may be directly transduced or transfected with GSE in vivo.

One embodiment of the present invention are GSEs that overlap with the HIV gene rev and the use of such GSEs to prevent tumorigenesis.

Claim 1 of 6 Claims

What is claimed is:

1. A method of inhibiting tumorigenesis, comprising contacting a cell with an isolated polynucleotide comprising a nucleic acid sequence which hybridizes under highly stringent conditions to a nucleic acid comprising a nucleotide sequence complementary to any one of SEQ ID NO:15 or SEQ ID NO:16, wherein said isolated polynucleotide is operably linked to a regulatory sequence, and expression of said isolated polynucleotide in a host cell inhibits hdm2 translocation.

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