Analysis of DNA-bound advanced glycation end-products by LC and mass spectrometry

Bidmon, Clemens; Frischmann, Matthias; Pischetsrieder, Monika

doi:10.1016/j.jchromb.2006.11.033

Cited by 20 publications

(12 citation statements)

References 72 publications

(92 reference statements)

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“…This observation in addition to the fact that all nucleated cells possess the same DNA content suggests that measurement of MG-dG adducts may provide a quantitative indicator of local (tissue DNA) or systemic (blood, urine) exposure to MG. The major thermodynamically stable DNA-AGE in biological samples resulting from reaction with MG is CEdG; we and others have proposed it as a general indicator of carbonyl stress and a specific surrogate biomarker for MG exposure [ 25 , 39 , 41 , 42 , 43 ]. We have described a quantitative and sensitive Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometry (LC-ESI-MS/MS) method using isotope dilution for the measurement of CEdG in biological samples [ 25 ].…”

Section: Resultsmentioning

confidence: 99%

Inhibition of GLO1 in Glioblastoma Multiforme Increases DNA-AGEs, Stimulates RAGE Expression, and Inhibits Brain Tumor Growth in Orthotopic Mouse Models

Jandial

Neman

Lim

et al. 2018

IJMS

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Cancers that exhibit the Warburg effect may elevate expression of glyoxylase 1 (GLO1) to detoxify the toxic glycolytic byproduct methylglyoxal (MG) and inhibit the formation of pro-apoptotic advanced glycation endproducts (AGEs). Inhibition of GLO1 in cancers that up-regulate glycolysis has been proposed as a therapeutic targeting strategy, but this approach has not been evaluated for glioblastoma multiforme (GBM), the most aggressive and difficult to treat malignancy of the brain. Elevated GLO1 expression in GBM was established in patient tumors and cell lines using bioinformatics tools and biochemical approaches. GLO1 inhibition in GBM cell lines and in an orthotopic xenograft GBM mouse model was examined using both small molecule and short hairpin RNA (shRNA) approaches. Inhibition of GLO1 with S-(p-bromobenzyl) glutathione dicyclopentyl ester (p-BrBzGSH(Cp)2) increased levels of the DNA-AGE N2-1-(carboxyethyl)-2′-deoxyguanosine (CEdG), a surrogate biomarker for nuclear MG exposure; substantially elevated expression of the immunoglobulin-like receptor for AGEs (RAGE); and induced apoptosis in GBM cell lines. Targeting GLO1 with shRNA similarly increased CEdG levels and RAGE expression, and was cytotoxic to glioma cells. Mice bearing orthotopic GBM xenografts treated systemically with p-BrBzGSH(Cp)2 exhibited tumor regression without significant off-target effects suggesting that GLO1 inhibition may have value in the therapeutic management of these drug-resistant tumors.

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Section: Resultsmentioning

confidence: 99%

Inhibition of GLO1 in Glioblastoma Multiforme Increases DNA-AGEs, Stimulates RAGE Expression, and Inhibits Brain Tumor Growth in Orthotopic Mouse Models

Jandial

Neman

Lim

et al. 2018

IJMS

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“…However, we are interested to further research by investigating the effect of repeated small doses of MG, the nature of the TNF‐α protein/message dichotomy and the result of cytokine loss on the differentiation of T cells after stimulation by MG‐exposed APC. It will also be important to identify key targets of MG damage, in cells of the immune system and elsewhere, in order to further knowledge of MG‐associated disease and to provide direction for therapeutic advances: MG reacts with arginine, lysine and cysteine residues of proteins, as well as with guanine units in DNA [25]. Depending upon the configuration and environment of proteins, some residues will be more prone to attack from MG than others, creating ‘hot spots’ for glycation [26, 27].…”

Section: Discussionmentioning

confidence: 99%

Methylglyoxal modulates immune responses: relevance to diabetes

Price

Hassi

English

et al. 2010

J Cellular Molecular Medi

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Increased methylglyoxal (MG) concentrations and formation of advanced glycation end-products (AGEs) are major pathways of glycaemic damage in diabetes, leading to vascular and neuronal complications. Diabetes patients also suffer increased susceptibility to many common infections, the underlying causes of which remain elusive. We hypothesized that immune glycation damage may account for this increased susceptibility. We previously showed that the reaction mixture (RM) for MG glycation of peptide blocks up regulation of CD83 in myeloid cells and inhibits primary stimulation of T cells. Here, we continue to investigate immune glycation damage, assessing surface and intracellular cytokine protein expression by flow cytometry, T-cell proliferation using a carboxyfluorescein succinimidyl ester assay, and mRNA levels by RT-PCR. We show that the immunomodulatory component of this RM was MG itself, with MG alone causing equivalent block of CD83 and loss of primary stimulation. Block of CD83 expression could be reversed by MG scavenger N-acetyl cysteine. Further, MG within RM inhibited stimulated production of interleukin (IL)-10 protein from myeloid cells plus interferon (IFN)-γ and tumour necrosis factor (TNF)-α from T cells. Loss of IL-10 and IFN-γ was confirmed by RT-PCR analysis of mRNA, while TNF-α message was raised. Loss of TNF-α protein was also shown by ELISA of culture supernatants. In addition, MG reduced major histocompatibility complex (MHC) class I expression on the surface of myeloid cells and increased their propensity to apoptose. We conclude that MG is a potent suppressor of myeloid and T-cell immune function and may be a major player in diabetes-associated susceptibility to infection.

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“…1A). The sensitivity was greatly increased by the use of ammonium formate to support ESI and by mass analysis in the negative mode [21].…”

Section: Lc-ms ⁄ Ms Analysis Of Cedg Abmentioning

confidence: 99%

Analysis and biological relevance of advanced glycation end‐products of DNA in eukaryotic cells

et al. 2008

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Advanced glycation end‐products (AGEs) of DNA are formed spontaneously by the reaction of carbonyl compounds such as sugars, methylglyoxal or dihydroxyacetone in vitro and in vivo. Little is known, however, about the biological consequences of DNA AGEs. In this study, a method was developed to determine the parameters that promote DNA glycation in cultured cells. For this purpose, the formation rate of N2‐carboxyethyl‐2′‐deoxyguanosine (CEdG), a major DNA AGE, was measured in cultured hepatic stellate cells by liquid chromatography (LC)‐MS/MS. In resting cells, a 1.7‐fold increase of CEdG formation rate was observed during 14 days of incubation. To obtain insights into the functional consequences of DNA glycation, CEdG was introduced into a luciferase reporter gene vector and transfected into human embryonic kidney (HEK 293 T) cells. Gene activity was determined by chemiluminescence of the luciferase. Thus, CEdG adducts led to a dose‐dependent and highly significant decrease in protein activity, which is caused by loss of functionality of the luciferase in addition to reduced transcription of the gene. When the CEdG‐modified vector was transformed into Escherichia coli, a loss of ampicillin resistance was observed in comparison to transformation with the unmodified plasmid. These results indicate that CEdG accumulates in the genomic DNA of resting cells, which could lead to diminished protein activity.

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Analysis of DNA-bound advanced glycation end-products by LC and mass spectrometry

Cited by 20 publications

References 72 publications

Inhibition of GLO1 in Glioblastoma Multiforme Increases DNA-AGEs, Stimulates RAGE Expression, and Inhibits Brain Tumor Growth in Orthotopic Mouse Models

Inhibition of GLO1 in Glioblastoma Multiforme Increases DNA-AGEs, Stimulates RAGE Expression, and Inhibits Brain Tumor Growth in Orthotopic Mouse Models

Methylglyoxal modulates immune responses: relevance to diabetes

Analysis and biological relevance of advanced glycation end‐products of DNA in eukaryotic cells

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