Method for determination and quantification of radiation or genotoxin exposure
The present invention discloses methods for detecting exposure of a living subject to genotoxic agents, testing sensitivity to a genotoxic agent, and determining DNA damage caused by exposure to an agent, comprising detecting the presence of FANCD2-containing foci from a sample collected from said subject. The presence of concentrated foci is indicative of DNA damage, and the degree of foci formation is correlated with degree of exposure. Diagnostic reagents contain a ligand that binds to human FANCD2 associated with a detectable label. Kits for detecting DNA damage in a biological sample contain such diagnostic reagents and signal detection components. The invention further discloses methods for identifying agents which modulate the ability of FANCD2-containing foci to form. Among other things, such agents are potentially useful chemosensitizing agents or may confer protection against damage caused by genotoxic agents.
1. A method of determining a subject's sensitivity to a genotoxic agent, comprising:
a) exposing a cancer cell obtained from said subject having a cancer to said genotoxic agent, and
b) detecting FANCD2-containing foci in said cancer cell,
wherein the presence or increase of said FANCD2-containing foci in said cancer cell compared to a control cell not exposed to said genotoxic agent indicates said subject's sensitivity to said genotoxic agent.
2. The method of claim 1 , wherein said cancer cell is obtained from said subject prior to, during or after said subject having a treatment for said cancer.
3. The method of claim 1 , wherein said cancer cell contains FANCD2-containing foci prior to exposure to said genotoxic agent.
4. The method of claim 1 , wherein said control cell is a cancer cell.
5. The method of claim 1 , wherein said control cell is obtained from said subject prior to said subject receiving a treatment for said cancer.
6. The method of claim 1 , wherein said FANCD2-containing foci are detected with an antibody specific for FANCD2.
7. The method of claim 6 , wherein said FANCD2-containing foci are detected by immunological methods selected from immunohistochemistry and immunofluorescence.
8. The method of claim 1 , wherein said cancer cell is obtained from a primary tumor site, a metastatic tumor site, a lymph node, a bodily fluid or blood.
9. A method of determining a subject's resistance to a genotoxic agent, comprising:
a) exposing a cancer cell obtained from said subject having a cancer to said genotoxic agent, and
b) detecting FANCD2-containing foci in said cancer cell,
wherein the absence of said FANCD2-containing foci or the lack of difference in FANCD2-foci formation in said cancer cell compared to a control cell not exposed to said genotoxic agent indicates said subject's resistance to said genotoxic agent.
10. The method of claim 9 , wherein said cancer cell is obtained from said subject prior to, during or after said subject having a treatment for said cancer.
11. The method of claim 9 , wherein said cancer cell contains FANCD2-containing foci prior to exposure to said genotoxic agent.
12. The method of claim 9 , wherein said control cell is a cancer cell.
13. The method of claim 9 , wherein said control cell is obtained from said subject prior to said subject receiving a treatment for said cancer.
14. The method of claim 9 , wherein said FAND2-containing foci are detected with an antibody specific for FANCD2.
15. The method of claim 14 , wherein said FANCD2-containing foci are detected by immunological methods selected from immunohistochemistry and immunofluorescence.
16. The method of claim 9 , wherein said cancer cell is obtained from a primary tumor site, a metastatic tumor site, a lymph node, a bodily fluid or blood.
17. A method of assessing whether a cell has a functional Fanconi Anemia (FA) pathway, comprising:
a) exposing a test cell to a genotoxic agent, and
b) detecting FANCD2-containing foci in said test cell, wherein the presence or increase of said FANCD2-containing foci in said test cell compared to a control cell not exposed to said genotoxic agent indicates that said test cell has a functional FA pathway,
wherein the absence of FANCD2-containing foci or lack of difference in FANCD2-foci formation in said test cell compared to a control cell not exposed to said genotoxic agent indicates that said test cell does not have a functional FA pathway, and wherein a functional FA pathway is capable of forming FANCD2-containing foci.
18. The method of claim 17 , wherein said test cell is a cancer cell.
19. The method of claim 18 , wherein said cancer cell is obtained from a primary tumor site, a metastatic tumor site, a lymph node, a bodily fluid or blood.
20. The method of claim 17 , wherein said test cell has not been previously exposed to said genotoxic agent.
21. The method of claim 17 , wherein said test cell has been previously exposed to said genotoxic agent.
22. The method of claim 17 , wherein said test cell contains FANCD2-containing foci prior to exposure to said genotoxic agent.
23. The method of claim 17 , wherein said FAND2-containing foci are detected with an antibody specific for FANCD2.
24. The method of claim 23 , wherein said FAND2-containing foci are detected by immunological methods selected from immunohistochemistry and immunofluorescence.