Stress-responsive induction of a therapeutic agent and methods of use
This invention relates to compositions and methods for selective expression of a heterologous nucleic acid sequence in a targeted tissue, and more particularly to the glucose regulated protein 78 (grp78) stress-responsive promoter and its use in gene therapy and the production of transgenic animals.
1. A method for inhibiting cell proliferation associated with glucose starvation comprising:
a) transducing a target cell capable of cell proliferation with a recombinant retroviral vector comprising a nucleic acid construct, the nucleic acid construct comprising (i) at least one glucose responsive protein 78 (grp78) non-coding regulatory sequence that comprises a sequence from about 520 base pairs 5′ of the site of initiation of transcription of the rat grp78coding sequence to about 175 base pairs 3′ of the site of initiation of transcription of the rat grp78 coding sequence; and (ii) a heterologous nucleic acid sequence operatively linked to the regulatory sequence, wherein the heterologous sequence comprises a structural gene that encodes a biologically active enzyme that converts a non-therapeutically effective compound to a therapeutically-effective compound in vivo;
b) activating the glucose responsive protein 78 (grp78) non-coding regulatory sequence such that the heterologous nucleic acid sequence comprising a structural gene that encodes a biologically active protein is expressed; and
c) contacting the cell with a non-therapeutically effective compound that is subsequently converted to a therapeutically-effective compound by the biologically active protein,
wherein the therapeutically effective compound inhibits cell proliferation associated with glucose starvation.
2. The method of claim 1 , wherein the subject is a mammal.
3. The method of claim 2 , wherein the mammal is a mouse.
4. The method of claim 2 , wherein the mammal is a human.
5. The method of claims 1 , wherein the administration is by in vivo administration.
6. The method of claim 5 , wherein the in vivo administration is by direct administration.
7. The method of claim 1 , wherein the cell proliferative disorder is a neoplastic disorder.
8. The method of claim 7 , wherein the neoplastic disorder is selected from the group consisting of lung cancer, colon-rectum cancer, breast cancer, prostate cancer, urinary tract cancer, uterine cancer lymphoma, oral cancer, pancreatic cancer, leukemia, melanoma, stomach cancer, thyroid cancer, liver cancer, and brain cancer and ovarian cancer.