Measurement of the malondialdehyde–2′‐deoxyguanosine adduct in human urine by immuno‐extraction and liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry

Hoberg, Anne-Mette; Otteneder, Michael B.; Marnett, Lawrence J.; Poulsen, Henrik E.

doi:10.1002/jms.547

Cited by 50 publications

(55 citation statements)

References 21 publications

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“…This may explain the reason that intracellular levels of M 1 dG are relatively insensitive to variations in plasma PUFA [33] although oxidation of PUFAs is known to give MDA and M 1 dG [33]. Furthermore, M 1 dG has been detected in urine samples at levels of 12+/-3.8 fmol kg -1 [35] and may also be oxidized to 6-oxo-M 1 dG adducts prior to excretion in urine [36,37]. It therefore appears that analysis of M 1 dG from blood DNA will not be a useful biomarker of colorectal cancer risk due to rapid excision and excretion and that the analysis of M 1 dG adducts in urine may prove to be more beneficial.…”

Section: Discussionmentioning

confidence: 99%

The Effect of n-6 Polyunsaturated Fatty Acid on Blood Levels of Malondialdehyde-Deoxyguanosine Adducts in Human Subjects

Moore¹,

Humphreys²,

Friesen³

et al. 2008

TOBIOMJ

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Abstract:The role of n-6 polyunsaturated fats upon the formation of the mutagenic DNA adduct malondialdehydedeoxyguanosine (M 1 dG) in blood was investigated in male volunteers (n = 13) who consumed diets high in saturated and polyunsaturated fats, and polyunsaturated fat plus a-tocopherol supplemention (400 IU per day). On day 14 there was a significant difference in adduct levels between diets with saturated fats giving higher levels than polyunsaturated fats but this effect had disappeared by day 20 indicating that there is a relatively rapid adjustment to the effects on DNA damage of changes in dietary fat. a-Tocopherol showed a small benefit by day 20. Five females participated in the PUFA study and had higher mean adduct levels than men but there was no correlation with hormonal status. Overall, PUFA had a limited beneficial effect on M 1 dG levels that warrants further investigation.

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Section: Discussionmentioning

confidence: 99%

The Effect of n-6 Polyunsaturated Fatty Acid on Blood Levels of Malondialdehyde-Deoxyguanosine Adducts in Human Subjects

Moore¹,

Humphreys²,

Friesen³

et al. 2008

TOBIOMJ

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“…The oxidized guanine adducts, 8-oxo-G and 8-oxo-dG, have been reported to be present in the urine of healthy human donors at levels of Ϸ2 nmol͞kg per 24 h and 0.4 nmol͞kg per 24 h, respectively (18). Attempts to quantify M 1 dG indicated that it is present at much lower concentrations in human urine, raising the possibility that it is metabolized before excretion (17).…”

Section: Discussionmentioning

confidence: 99%

“…We recently described a highly sensitive and specific method based on immunoaffinity purification and liquid chromatography (LC)-MS (16,17). Application of this technique reveals that M 1 dG is detectable in the urine of healthy human volunteers in amounts corresponding to 10-20 fmol͞kg per 24 h (17).…”

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confidence: 99%

In vivo oxidative metabolism of a major peroxidation-derived DNA adduct, M ₁ dG

Otteneder¹,

Knutson²,

Daniels³

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

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3-(2-Deoxy-␤-D-erythro-pentofuranosyl)pyrimido[1,2-␣]purin-10(3H)-one (M1dG) is a DNA adduct arising from the reaction of 2-deoxyguanosine with the lipid peroxidation product, malondialdehyde, or the DNA peroxidation product, base propenal. M 1dG is mutagenic in bacteria and mammalian cells and is present in the genomic DNA of healthy human beings. It is also detectable, albeit at low levels, in the urine of healthy individuals, which may make it a useful biomarker of DNA damage linked to oxidative stress. We investigated the possibility that the low urinary levels of M 1dG reflect metabolic conversion to derivatives. M 1dG was rapidly removed from plasma (t1/2 ‫؍‬ 10 min) after i.v. administration to rats. A single urinary metabolite was detected that was identified as 6-oxo-M 1dG by MS, NMR spectroscopy, and independent chemical synthesis. 6-Oxo-M1dG was generated in vitro by incubation of M 1dG with rat liver cytosols, and studies with inhibitors suggested that xanthine oxidase and aldehyde oxidase are involved in the oxidative metabolism. M1dG also was metabolized by three separate human liver cytosol preparations, indicating 6-oxo-M1dG is a likely metabolite in humans. This represents a report of the oxidative metabolism of an endogenous DNA adduct and raises the possibility that other endogenous DNA adducts are metabolized by oxidative pathways. 6-Oxo-M1dG may be a useful biomarker of endogenous DNA damage associated with inflammation, oxidative stress, and certain types of cancer chemotherapy.excretion ͉ inflammation ͉ DNA damage ͉ oxidation ͉ metabolite D NA damage from endogenous sources is believed to contribute significantly to human genetic diseases, including cancer (1, 2). A major cause of endogenous DNA damage is oxidation (3, 4). Agents such as hydroxyl radical or peroxynitrous acid oxidize nucleic acid bases or the deoxyribose backbone to form miscoding lesions or induce strand breaks (5). In addition, oxidation of other cellular constituents (i.e., lipid, protein) generates reactive derivatives that form adducts with nucleic acid bases (1). In the absence of repair, this panoply of damage results in mutations, triggers signaling to arrest cell division, or induces apoptosis.3-(2-Deoxy-␤-D-er ythro-pentofuranosyl)pyrimido[1,2-␣]purin-10(3H)-one (M 1 dG) is an adduct found at varying levels in genomic DNA of rodents and humans (6-8). It is derived from the lipid oxidation product, malondialdehyde, or the DNA oxidation product, base propenal (Scheme 1) (9-11). M 1 dG is miscoding when assayed by in vitro DNA replication and is mutagenic in bacterial and mammalian cells (12)(13)(14). It induces base pair substitutions (M 1 dG3T and M 1 dG3A) and frameshift mutations in reiterated sequences (e.g., CG n ) when shuttle vectors containing a site-specific lesion are replicated in COS-7 cells (14). Genetic and biochemical experiments indicate that M 1 dG is removed by nucleotide excision repair in both bacterial and mammalian cells (13-15).As a prerequisite for preclinical, clinical, and populationbased ...

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“…M 1 -dG is reported to be a substrate for the nucleotide excision repair pathway (NER), which accounts for its appearance in human urine (13,20). This repair-dependent removal and subsequent deposition into urine provides an available source of M 1 dG for monitoring the exposure to oxidative damage (21).…”

Section: Introductionmentioning

confidence: 99%

Metabolism in Vitro and in Vivo of the DNA Base Adduct, M₁G

Knutson

Akingbade

Crews

et al. 2007

Chem. Res. Toxicol.

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Oxidative damage is considered a major contributing factor to genetic diseases including cancer. Our laboratory is evaluating endogenously formed DNA adducts as genomic biomarkers of oxidative injury. Recent efforts have focused on investigating the metabolic stability of adducts in vitro and in vivo. Here, we demonstrate that the base adduct, M 1 G, undergoes oxidative metabolism in vitro in rat liver cytosol (RLC, K m ) 105 µM and V max /K m ) 0.005 min -1 mg -1 ) and in vivo when administered intravenously to male Sprague Dawley rats. LC-MS analysis revealed two metabolites containing successive additions of 16 amu. One-and two-dimensional NMR experiments showed that oxidation occurred first at the 6-position of the pyrimido ring, forming 6-oxo-M 1 G, and then at the 2-position of the imidazole ring, yielding 2,6-dioxo-M 1 G. Authentic 6-oxo-M 1 G was chemically synthesized and observed to undergo metabolism to 2,6-dioxo-M 1 G in RLC (K m ) 210 µM and V max /K m ) 0.005 min -1 mg -1 ). Allopurinol partially inhibited M 1 G metabolism (75%) and completely inhibited 6-oxo-M 1 G metabolism in RLC. These inhibition studies suggest that xanthine oxidase is the principal enzyme acting on M 1 G in RLC and the only enzyme that converts 6-oxo-M 1 G to 2,6-dioxo-M 1 G. Both M 1 G and 6-oxo-M 1 G are better substrates (5-fold) for oxidative metabolism in RLC than the deoxynucleoside, M 1 dG. Alternative repair pathways or biological processing of M 1 dG makes the fate of M 1 G of interest as a potential marker of oxidative damage in vivo.

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Measurement of the malondialdehyde–2′‐deoxyguanosine adduct in human urine by immuno‐extraction and liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry

Cited by 50 publications

References 21 publications

The Effect of n-6 Polyunsaturated Fatty Acid on Blood Levels of Malondialdehyde-Deoxyguanosine Adducts in Human Subjects

The Effect of n-6 Polyunsaturated Fatty Acid on Blood Levels of Malondialdehyde-Deoxyguanosine Adducts in Human Subjects

In vivo oxidative metabolism of a major peroxidation-derived DNA adduct, M ₁ dG

Metabolism in Vitro and in Vivo of the DNA Base Adduct, M₁G

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Measurement of the malondialdehyde–2′‐deoxyguanosine adduct in human urine by immuno‐extraction and liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry

Cited by 50 publications

References 21 publications

The Effect of n-6 Polyunsaturated Fatty Acid on Blood Levels of Malondialdehyde-Deoxyguanosine Adducts in Human Subjects

The Effect of n-6 Polyunsaturated Fatty Acid on Blood Levels of Malondialdehyde-Deoxyguanosine Adducts in Human Subjects

In vivo oxidative metabolism of a major peroxidation-derived DNA adduct, M 1 dG

Metabolism in Vitro and in Vivo of the DNA Base Adduct, M1G

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In vivo oxidative metabolism of a major peroxidation-derived DNA adduct, M ₁ dG

Metabolism in Vitro and in Vivo of the DNA Base Adduct, M₁G