Therapeutic and diagnostic strategies
The present invention encompasses the finding that microRNAs (miRNAs) regulate certain key proteins involved in DNA repair. In some embodiments, a miRNA suppresses levels and/or activity of one or more DNA repair proteins. In some such embodiments, such suppression renders cells hypersensitive to certain DNA damage agents (e.g., γ-irradiation and genotoxic drugs, among others). The present invention provides various reagents and methods associated with these findings including, among other things, strategies for treating cell proliferative disorders, certain diagnostic systems, etc.
1. A method comprising steps of:
administering to an individual who is suffering from or susceptible to a cell proliferative disorder a composition comprising an miR-24 agent in an amount sufficient to inhibit cell proliferation in combination with a DNA damage agent and/or cell proliferation disorder treatment method that produces DNA double-strand breaks,
isolating cells from the individual administered the miR-24 agent and preparing Wright-stained metaphase chromosome spreads from the isolated cells or staining the isolated cells from the individual with SYBR Green and performing single cell gel electrophoresis on the isolated cells, thereby measuring DNA damage and detecting DNA damage repair to be reduced in the isolated cells, as compared to control cells, thereby identifying the individual administered the miR-24 agent to possess reduced DNA damage repair,
further comprising administering to the individual so identified to possess reduced DNA damage repair the miR-24 agent and γ-irradiation or a genotoxic drug.
2. A method comprising steps of:
administering to an individual who is suffering from or susceptible to a cell proliferative disorder a composition comprising an miR-24 agent in an amount sufficient to render cells of the individual hypersensitive to a DNA damage agent and/or cell proliferation disorder treatment method that produces DNA double-strand breaks,
isolating cells from the individual administered the miR-24 agent and preparing Wright-stained metaphase chromosome spreads from the isolated cells or staining the isolated cells from the individual with SYBR Green and performing single cell gel electrophoresis on the isolated cells, thereby measuring DNA damage and detecting DNA damage repair to be reduced in the isolated cells, as compared to control cells, thereby identifying the individual administered the miR-24 agent to possess reduced DNA damage repair,
further comprising administering to the individual so identified to possess reduced DNA damage repair the miR-24 agent and γ-irradiation or a genotoxic drug.
3. The method of claim 1 , wherein the amount is an amount sufficient to suppress levels and/or activity of one or more DNA repair proteins.
4. The method of claim 2 , wherein the DNA repair proteins are selected from the group consisting of photolyases, methyltransferases, AP endonucleases, exonucleases, histone variants, transcription factors that regulate expression of DNA repair genes, and DNA glycosylases, and combinations thereof.
5. The method of claim 1 , wherein the step of administering comprises administering in combination with other cell proliferation disorder treatment methods and/or therapeutic agents.
6. The method of claim 2 , wherein the amount is an amount sufficient to suppress levels and/or activity of one or more DNA repair proteins.
7. The method of claim 2 , wherein the step of administering comprises administering in combination with other cell proliferation disorder treatment methods and/or therapeutic agents.
8. The method of claim 1 or 2 , wherein the cell proliferative disorder is a hematopoietic cell proliferative disorder.
9. A method comprising steps of:
administering to an individual who is suffering from or susceptible to a cell proliferative disorder a composition comprising an miR-24 agent in an amount sufficient to down-regulate the expression of H2AFX, and thereby inhibit cell proliferation,
wherein the miR-24 agent is administered in combination with a DNA damage agent and/or cell proliferation disorder treatment method that produces DNA double-strand breaks,
isolating cells from the individual administered the miR-24 agent, preparing cDNA from the isolated cells and performing quantitative RT-PCR on the cDNA using primers from the H2AX coding region and SYBR Green, thereby measuring H2AFX expression and detecting H2AFX expression to be reduced in the isolated cells, as compared to control cells, thereby identifying the individual administered the miR-24 agent to possess reduced H2AFX expression,
further comprising administering to the individual so identified to possess reduced H2AFX expression the miR-24 agent and γ-irradiation or a genotoxic drug.