Therapeutic gene-switch constructs and bioreactors for the expression of biotherapeutic molecules, and uses thereof

Abstract: The present invention relates to methods and compositions for treating, ameliorating or preventing a disease or disorder in a subject by introducing into cells of the subject a therapeutic gene switch construct that controls expression of one or more therapeutic products. (end of abstract)

Therapeutic gene-switch constructs and bioreactors for the expression of biotherapeutic molecules, and uses thereof description/claims

Brief Patent Description

Full Patent Description

Patent Application Claims

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application claims the benefit of the filing date of U.S. Provisional Application No. 61/047,899, files Apr. 25, 2008 and U.S. Provisional Application No. 60/975,986, filed Sep. 28, 2007, both of which are incorporated herein by reference in their entireties.

REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY VIA EFS-WEB

The content of the electronically submitted sequence listing (Name: Sequence Listing.ST25.txt; Size: 243,000 bytes; and Date of Creation: Sep. 29, 2008) filed herewith the application is incorporated herein by reference in its entirety:

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to methods and compositions for treating, ameliorating, or preventing a disease, disorder, or condition in a subject by introducing into the subject a therapeutic gene switch construct that controls expression of one or more therapeutic products. In a further embodiment, the present invention relates to methods and compositions for treating, ameliorating, or preventing a disease, disorder, or condition in a subject by introducing into the subject a “bioreactor,” a therapeutic implant composed of a cell or cells that secrete a therapeutic protein. A bioreactor may be immuno-isolated by encapsulation or non-immunoisolated. In particular embodiments, the bioreactor comprises a therapeutic gene switch construct.

2. Background of the Invention

The concept of treating or preventing a disease in a subject through introduction of a polynucleotide encoding a therapeutic molecule, e.g., a therapeutic polypeptide or therapeutic polynucleotide into cells of the subject, or introducing into the subject modified cells engineered to secrete the therapeutic molecule has been in existence for many years. Several difficulties in the practical aspects of the concept have hindered progress towards successful therapies. Direct introduction of genetic material into a subject to be treated presents difficulties such as: safety of delivery, obtaining sufficient expression levels of the therapeutic product for a sufficient period of time, limiting expression of the therapeutic product to desired cells, and maintaining the ability to modulate or pulse the expression of the therapeutic product, including the ability to turn off expression of the therapeutic product if it is no longer needed. Cell based therapies are subject to rejection via the subject\'s immune response, therefore immuno-isolation strategies such as cell encapsulation methods have been developed to increase the longevity of implanted cells and allow use of xenogeneic cells, i.e., cells from a different species. Current encapsulated and non-encapsulated cell therapies are engineered to secrete the therapeutic protein constitutively. Once implanted, protein secretion can not be regulated. To improve the safety and clinical application of direct or cell-mediated bioreactor therapeutic protein delivery it would be advantageous to be able to turn off the protein production or regulate the rate at which protein production occurs.

Thus, there is a need in the art for new therapeutic methods and compositions that provide these desired characteristics.

SUMMARY OF THE INVENTION

The present invention relates to methods and compositions for treating, ameliorating, or preventing a disease, disorder, or condition in a subject.

In one embodiment, the present invention provides a method for treating, ameliorating, or preventing a disease, disorder, or condition in a subject, comprising:

(a) introducing into a subject (1) a first polynucleotide encoding a gene switch, where the gene switch comprises at least one transcription factor sequence encoding a ligand-dependent transcription factor through operable association with a therapeutic switch promoter, where the therapeutic switch promoter is constitutively active and (2) a second polynucleotide encoding a therapeutic polypeptide or therapeutic polynucleotide in operable association with a factor-regulated promoter which is activated by said ligand-dependent transcription factor, where the first and second polynucleotides are introduced so as to permit their expression in the presence of ligand; and

(b) administering ligand to the subject to induce expression of the therapeutic polypeptide or therapeutic polynucleotide.

A further embodiment of the invention provides a method for expressing a therapeutic polypeptide or therapeutic polynucleotide in a subject, comprising:

- (a) introducing into a subject (1) a first polynucleotide encoding a gene switch, where the gene switch comprises at least one transcription factor sequence encoding a ligand-dependent transcription factor through operable association with a therapeutic switch promoter, where the therapeutic switch promoter is activated under conditions associated with the disease, disorder, or condition to be treated, and (2) a second polynucleotide encoding a therapeutic polypeptide or therapeutic polynucleotide in operable association with a factor-regulated promoter which is activated by said ligand-dependent transcription factor, where the first and second polynucleotides are introduced so as to permit their expression in the subject under conditions associated with the disease, disorder, or condition; and
- (b) administering ligand to the subject to induce expression of the therapeutic polypeptide or therapeutic polynucleotide.