Methods and systems for evaluating health risk factors by measurement of DNA damage and DNA repair

• US 20070269824A1
• Filed: 03/02/2007
• Published: 11/22/2007
• Est. Priority Date: 03/02/2006
• Status: Abandoned Application

• Current Assignee: Andrew Conrad, Francoise Gala, Jeffrey Albrecht, Russell Grant
• Original Assignee: Andrew Conrad, Francoise Gala, Jeffrey Albrecht, Russell Grant
• Inventors: Conrad, Andrew, Gala, Francoise, Albrecht, Jeffrey, Grant, Russell
• Application Number:
  US11/713,451
• Publication Number:
  US 20070269824A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/00 Measuring or testing processes involving enzymes, nucleic acids or microorganisms measuring or testing apparatus with condition measuring or sensing means, e.g. colony counters, C12M1/34; Compositions therefor; Processes of preparing such compositions
• CPC Class Codes: G01N 2333/90216 acting on a heme group of donors (1.9)
• CPC Class Codes: G01N 2333/91102 with definite EC number (2.4.1.-)
• CPC Class Codes: G01N 2333/922 Ribonucleases (RNAses); Deoxyribonucleases (DNAses)
• CPC Class Codes: G01N 33/5017 for testing neoplastic activity

Abstract

Disclosed are methods for measuring the effects of environmental, physiological, or lifestyle variables on DNA damage and DNA repair activity as well as the use of measurements of DNA damage and DNA repair activity to predict increased risk for disease. Embodiments of the methods involve the use of a combination of assays to measure DNA damage and DNA repair activity in an individual and comparing these measurements to suitable controls using the selected assays for normal healthy individuals of varying ages. In other embodiments, the methods may comprise a comparison of DNA damage levels to DNA repair levels to obtain an apparent net measurement of DNA damage accumulation.

Claims

1. A method to correlate the effect of at least one variable on at least one of DNA damage or DNA repair in an individual comprising the steps of:

(a) using a plurality of assays to measure at least one of DNA damage or DNA repair in the individual;

(b) determining whether the amount of at least one of DNA damage or DNA repair as measured in the individual differs from the amount of at least one of DNA damage or DNA repair for a plurality of control samples; and

(c) determining if there is a correlation between the levels of at least one of DNA damage or DNA repair in the individual to changes in the variable.

2. The method of claim 1, wherein the variable correlated to at least one of DNA damage or DNA repair comprises at least one of an environmental variable, a lifestyle variable, or a physiological variable.

3. The method of claim 1, wherein a positive correlation between an increase in DNA damage and the at least one variable provides an indication that the at least one variable may be associated with damage to the individual'"'"'s DNA.

4. The method of claim 1, wherein a correlation between an increase in DNA repair and the at least one variable provides an indication that the at least one variable may be associated with damage to the individual'"'"'s DNA.

5. The method of claim 1, wherein step (b) comprises measurement of both DNA repair and DNA damage by a plurality of assays for both DNA repair and DNA damage.

6. The method of claim 1, wherein an increase in the level of DNA damage that is not coupled with a comparable increase in the level of DNA repair, or a decrease in the level of DNA repair that is not coupled with a comparable decrease in the level of DNA damage, for the individual as compared to the plurality of controls, is indicative of a negative impact of the at least one variable on overall DNA damage accumulation in the individual.

7. The method of claim 1, wherein the control samples provide a range of reference values for the assay being used to monitor DNA damage or DNA repair.

8. The method of claim 1, wherein the control samples comprise samples taken from healthy subjects that are of a similar age as the tested individual.

9. The method of claim 1, wherein the control samples comprise samples taken from subjects that are exposed to the same variable or variables as the tested individual.

10. The method of claim 1, wherein the measurement of DNA damage is quantified by a DNA adduct assay.

11. The method of claim 10, wherein the DNA adduct assay comprises the measurement of DNA adducts using at least one of high performance liquid chromatography in combination with electrochemical detection (HPLC-ECD) or two-dimensional liquid chromatography in combination with tandem mass spectrometry (2D-LC-MS/MS).

12. The method of claim 10, wherein the DNA adduct assay measures the level of a DNA adduct comprising at least one of 8-hydroxydeoxyguanosine (8OHdG), 8-hydroxyguanosine (8OHG), 8-oxoguanine (8-oxo-G), 2,6-Diamino-4-hydroxy-5-formamidopyrimidine (FapyGua), 8-hydroxy-adenine/8-oxoadenine (8OHAde), O6-methyl-guanine, 4,6-Diamino-5-formamidopyrimidine (FapyAde), 5-hydroxy-cytosine (5-OH-Cyt), 5-Hydroxy-methylhydantoin (5-OH-5-MeHyd), 5-hydroxy-hydantoin (5-OH-Hyd), 2-oxoadenine (2-OH-Ade), or 5-Hydroxy-methyl-uracil (5-OH-Me-Ura).

13. The method of claim 1, wherein the measurement of DNA damage is quantified by a DNA break assay.

14. The method of claim 13, wherein the DNA break assay comprises at least one of a Comet Assay, an Aldehyde Reactive Probe Assay, or a DNA Ladder Assay.

15. The method of claim 1, wherein the measurement of DNA repair is quantified by at least one of a DNA Repair Enzyme assay, a Repair Capacity Analysis assay, or a DNA Damage Susceptibility assay.

16. The method of claim 15, wherein the DNA Repair Enzyme assay measures expression of at least one of 8-oxoguanine DNA glycosylase (OGG1), MutY homolog (hMYH), MutT Homolog-1 (MTH1), Heme oxygenase 1 (HOX1), NEIL endonuclease VIII-like 1 protein (NEIL1 protein), Nth homolog 1 (NTH1 protein), excision repair cross-complementing protein (ERCC1), AP endonuclease (Ape-1), or superoxide dismutase (SOD-1).

17. The method of claim 1, wherein the at least one variable being correlated to DNA damage or DNA repair comprises at least one of diet, physical activity, aging, pregnancy, stress, smoking, alcohol consumption, disease, disease treatment, drug treatment, antioxidant supplementation, cosmetic treatment, exposure to a carcinogen, or exposure to radiation.

18. The method of claim 17, wherein the cosmetic treatment is a chemical peel or laser resurfacing.

19. The method of claim 17, wherein the exposure to radiation involves exposure to X-ray radiation or ultraviolet radiation.

28. A kit to perform the method of claim 1.

20. A method to correlate effect of at least one of DNA damage or DNA repair to an increased or a decreased risk for disease in an individual comprising the steps of:

(a) using a plurality of assays to measure at least one of DNA damage or DNA repair in the individual;

(b) comparing the level of at least one of DNA damage or DNA repair in the individual to the control samples;

(c) determining if the amount of at least one of DNA damage or DNA repair as measured in the individual differs from the amount of at least one of DNA damage or DNA repair for a plurality of control samples; and

(d) determining if there is a correlation between a change in at least one of DNA damage or DNA repair and a change in the risk of the disease for the individual.

21. The method of claim 20, wherein step (b) comprises measurement of both DNA repair and DNA damage by a plurality of assays for both DNA repair and DNA damage.

22. The method of claim 21, wherein an increase in the relative level of DNA damage coupled with a decrease in the relative level of DNA repair for the individual as compared to the plurality of controls is correlated with an increased risk of at least one disease that is associated with DNA damage.

23. The method of claim 21, wherein an decrease in the relative level of DNA damage coupled with a increase in the relative level of DNA repair for the individual as compared to the plurality of controls is correlated with an decrease risk of at least one disease that is associated with DNA damage.

24. The assay of claim 20, wherein the control samples provide a range of reference values for the assay being used to monitor DNA damage or DNA repair.

25. The assay of claim 20, wherein the disease comprises a disease of oxidative stress or DNA degradation.

26. The assay of claim 20, wherein the disease is cancer or a degenerative disease.

27. A method to correlate effect of at least one of DNA damage or DNA repair to an increased or a decreased risk for disease in an individual comprising the steps of:

(a) using a plurality of assays to measure at least one of DNA damage and DNA repair in the individual at an initial time;

(b) repeating the measurements of step (a) at one or more later time points measuring at least one of DNA damage and DNA repair in the individual at a later time;

(c) determining whether the amount of at least one of DNA damage or DNA repair as measured for at least one later time point differs from the amount of at least one of DNA damage or DNA repair at the initial time point; and

(d) determining if there is a correlation between a change in at least one of DNA damage or DNA repair and a change in the risk of the disease for the individual.