Methods for determining and modulating cellular redox potential and genotoxic states

A method of screening for or prognosing cancer comprising the steps of:

• a) administering a surrogate compound to a test subject prior to obtaining a specimen therefrom wherein the compound exhibits a redox ambivalent state subsequent to an oxygen radical attack and forms at least two identifiable stable products;
• b) obtaining a specimen from the test subject wherein the specimen is known to contain detectible quantities of the surrogate compound;
• c) isolating the compound and stable products from the specimen;
• d) assaying for at least one stable product derived via reductive pathways and for at least one stable product derived via oxidative pathways wherein the at least one stable product derived via reductive pathways is associated with a redox potential favoring a reductive state and the at least one stable product derived via oxidative pathways is associated with a redox potential favoring an oxidative state;
• and
• e) identifying an increased risk of cancer or increased probability of its manifestation in the test specimen when the cellular redox potential favors an oxidative state or identifying a decreased risk of cancer or decreased probability of its manifestation in the test specimen when the cellular redox potential favors a reductive state.

A method for monitoring the efficacy of cellular redox modulation comprising the steps of:

• a) obtaining a specimen containing DNA from a test subject;
• b) isolating the DNA from the specimen;
• c) assaying for at least one modified nucleotide base derived via reductive pathways, and for at least one modified nucleotide base derived via oxidative pathways;
• d) comparing the concentration level of the at least one modified nucleotide base derived via reductive pathways to the concentration level of the at least one oxidatively modified nucleotide base derived via oxidative pathways to obtain a first data set;
• e) repeating steps a) through d) to obtain a second data set;
• and
• f) identifying a shift towards a reductive redox potential when the concentration level of the at least one modified nucleotide base derived via reductive pathways as observed in the second data set is greater than the concentration level of the at least one modified nucleotide base derived via oxidative pathways as observed in the first data set.

A method for determining the cellular redox potential of a specimen comprising the steps of:

• a) obtaining a specimen having cellular DNA from a test subject;
• b) isolating the DNA from the specimen;
• c) assaying the DNA for concentration levels of at least one modified nucleotide base derived via reductive pathways and for at least one modified nucleotide base derived via oxidative pathways;
• and
• d) comparing the concentration level of the at least one modified nucleotide base derived via reductive pathways to the concentration level of the at least one oxidatively modified nucleotide base derived via oxidative pathways, an oxidative redox potential existing when formation of modified bases derived via oxidative pathways are favored over modified bases derived via reductive pathways.

A method for modulating the cellular redox potential of a targeted cell group and monitoring the same comprising the steps of:

• a) introducing a gene expression activator into a test subject wherein the gene encodes a free radical detoxifying enzyme;
• b) obtaining a test specimen from the test subject known to have the targeted cells;
• c) isolating DNA from the specimen;
• d) assaying the DNA for concentration levels of at least one modified nucleotide base derived via reductive pathways and for at least one modified nucleotide base derived via oxidative pathways;
• and
• e) comparing the concentration level of the at least one modified nucleotide base derived via reductive pathways to the concentration level of the at least one oxidatively modified nucleotide base derived via oxidative pathways, wherein a decrease in the cellular redox potential occurs when formation of modified bases derived via oxidative pathways are disfavored over modified bases derived via reductive pathways.

A method for modulating the cellular redox potential of a targeted cell group and monitoring the same comprising the steps of:

• a) determining the gene sequence for a free radical detoxifying enzyme;
• b) preparing and cloning cDNA based upon the gene sequence;
• c) introducing the cDNA into a test subject;
• d) obtaining a test specimen from the test subject known to have been exposed to the cDNA;
• e) isolating DNA from the specimen;
• f) assaying the DNA for concentration levels of at least one modified nucleotide base derived via reductive pathways and for at least one modified nucleotide base derived via oxidative pathways;
• and
• g) comparing the concentration level of the at least one modified nucleotide base derived via reductive pathways to the concentration level of the at least one oxidatively modified nucleotide base derived via oxidative pathways, wherein a decrease in the cellular redox potential occurs when formation of modified bases derived via oxidative pathways are disfavored over modified bases derived via reductive pathways.