Abstract
The present invention provides methods useful in autologous bone marrow transplantation and cancer therapy. According to one aspect of the invention, bone marrow cells from a patient having cancer are treated with selected antisense oligonucleotides in order to deplete the bone marrow of malignant cells prior to infusion back into the bone marrow donor. In a separate embodiment, selected antisense oligonucleotides are administered systemically for anticancer therapy.
Patent number: 5087617
Filing date: Feb 15, 1989
Issue date: Feb 11, 1992
Inventor: Larry J. Smith
Assignee: Board of Regents, The University of Texas System
International Classification
A61K 3170
1. A method for treating bone marrow cells from an individual having cancer prior to infusion of the bone marrow cells back into the individual, comprising the steps of:
- a obtaining bone marrow cells from the individual; and
- b exposing the bone marrow cells to a proliferation inhibiting amount of an antisense oligonucleotide having a sequence complementary to a sequence of RNA transcribed from a target gene present in the cells of the cancer, wherein said target gene is selected from the group consisting of those genes encoding cell surface receptors, modulators of intracellular messengers, or transcriptional regulators.
2. A method for treating an individual having cancer comprising the steps of:
- a obtaining bone marrow cells from the individual; and
- b exposing the bone marrow cells to a proliferation inhibiting amount of an antisense oligonucleotide having a sequence complementary to a sequence of RNA transcribed from a target gene present in the cells of the cancer, wherein said target gene is selected from the group consisting of those genes encoding cell surface receptors, modulators of intracellular messengers, or transcriptional regulators; and
- c transplanting the exposed cells into the individual.
3. A method for treating bone marrow cells from an individual having cancer prior to infusion of the bone marrow cells back into the individual, comprising the steps of:
- a) obtaining bone marrow cells from the individual; and
- b) exposing the bone marrow cells to a proliferation inhibiting amount of an antisense oligonucleotide having a sequence complementary to a sequence of RNA transcribed from a target gene present in the cells of the cancer, wherein said target gene is selected from the group consisting of genes that encode a molecule that regulates cell proliferation or viability.
4. A method for treating an individual having cancer comprising the steps of:
- a) obtaining bone marrow cells from the individual; and
- b) exposing the bone marrow cells to a proliferation inhibiting amount of an antisense oligonucleotide having a sequence complementary to a sequence of RNA transcribed from a target gene present in the cells of the cancer, wherein said target gene is selected from the group consisting of genes that encode a molecule that regulates cell proliferation or viability.
- c) transplanting the exposed cells into the individual.
5. A method for treating bone marrow cells from an individual having cancer prior to infusion of the bone marrow cells back into the individual, comprising the steps of:
- a) obtaining bone marrow cells from the individual; and
- b) exposing the bone marrow cells to a proliferation inhibiting amount of an antisense oligonucleotide having a sequence complementary to a sequence of RNA transcribed from a target gene present in the cells of the cancer, wherein said target gene is a gene that encodes a molecule that regulates cellular differentiation.
6. A method for treating an individual having cancer comprising the steps of:
- a) obtaining bone marrow cells from the individual; and
- b) exposing the bone marrow cells to a proliferation inhibiting amount of an antisense oligonucleotide having a sequence complementary to a sequence of RNA transcribed from a target gene present in the cells of the cancer, wherein said target gene is a gene that encodes a molecule that regulates cellular differentiation; and
- c) transplanting the exposed cells into the individual.
7. The method of claim 1, 2, 3, 4, 5 or 6 wherein said oligonucleotide has sufficient complementarity with the target gene to form a duplex having a melting temperature of at least about 40 degrees Centigrade under physiologic conditions.
8. The method of claim 2 or 4 or 6 comprising the additional step of administering to the individual a sufficient amount of a preparation containing oligonucleotides complementary to RNA transcribed from a target gene present in the cells of the cancer to kill the cancerous cells of the individual.
9. The method of claim 1, 2, 3, 4, 5 or 6 where the cancer is further defined as a cancer of cells of the hemopoietic system.
10. The method of claim 1, 2, 3, 4, 5 or 6 where the cancer is leukemia.
11. The method of claim 1, 2, 3, 4, 5 or 6 where the cancer is myeloid leukemia.
12. The method of claim 1, 2, 3, 4, 5 or 6 where the cancer is lymphoma.
13. The method of claim 1, 2, 3, 4, 5 or 6 where the cancer is breast cancer.
14. The method of claim 1, 2, 3, 4, 5 or 6 where the cancer is multiple myeloma.
15. The method of claim 1, 2, 3, 4, 5 or 6 where the cancer is gastrointestinal cancer.
16. The method of claim 1, 2, 3, 4, 5 or 6 where the target is a traitor gene.
17. The method of claim 1, 2, 3, 4, 5 or 6 wherein the oligonucleotide has a charged backbone.
18. The method of claim 1, 2, 3, 4, 5 or 6 wherein the oligonucleotide has an uncharged backbone.
19. The method of claim 1, 2, 3, 4, 5 or 6 where the oligonucleotide is a methylphosphonate oligonucleotide.
20. The method of claim 1, 2, 3, 4, 5 or 6 where the oligonucleotide is a phosphorothioate oligonucleotide.
21. The method of claim 1, 2, 3, 4, 5 or 6 where the oligonucleotide comprises at least 8 bases and is complementary to a sequence of RNA located 5' to the initiation condon of said target.
22. The method of claim 1, 2, 3, 4, 5 or 6 where the oligonucleotide comprises the sequence 3'-GGTCTGACGGAAGGCCCAGTGACGGTAC-5'.
23. The method of claim 1, 2, 3, 4, 5 or 6 where the oligonucleotide comprises at least 8 bases and is located within 40 bases of the initiation codon.
24. The method of claim 1, 2, 3, 4, 5 or 6 where the proliferation-inhibiting amount is 10-200 micromolar.
25. The method of claim 1, 2, 3, 4, 5 or 6 where the exposing step comprises:
- a. fractionating the bone marrow cells to obtain a mononuclear cell fraction, and
- b. culturing the mononuclear fraction together with a proliferation inhibiting amount of the oligonucleotide for at least about 1-10 days.
26. A method for treating bone marrow cells from an individual having cancer prior to infusion of the bone marrow cells back into the individual, comprising the steps of:
- a) obtaining bone marrow cells from the individual;
- b) exposing the bone marrow cells to a proliferation inhibiting amount of an antisense oligonucleotide having a sequence complementary to a sequence of RNA transcribed from a target gene present in the cells of the cancer, wherein said target gene is p53.