Development and validation of a UPLC-UV method for the quantification of thiopurine methyltransferase enzyme activity in human erythrocytes

Illamola, Sílvia M.; Echaabi, A.K.; Mazeron, C.; Loriot, Marie‐Anne; Pallet, Nicolas

doi:10.1016/j.jchromb.2019.03.014

Cited by 6 publications

(4 citation statements)

References 27 publications

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“…There was no sign of haematological toxicity. A blood thiopurine metabolite assay was performed according to a previously described method, 7…”

Section: Case Reportmentioning

confidence: 99%

“…There was no sign of haematological toxicity. A blood thiopurine metabolite assay was performed according to a previously described method, 7 which revealed low 6‐TGN level (145 pmol/8 × 10 8 red blood cell count [RBC]; therapeutic level: 200–600 pmol/8 × 10 8 RBC) and dramatically increased 6‐MMPN level (>12 450 pmol/8 × 10 8 RBC; therapeutic level <5800 pmol/8 × 10 8 RBC), associated with an unfavourable [6‐MMPN:6‐TGN] metabolite ratio (86; favourable ratio 3–35). TPMT activity was slightly over the normal/high values at 16 nmol/h/mL of RBC (N: 8.5–15.5 nmol/h/mL of RBC).…”

Section: Case Reportmentioning

confidence: 99%

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Azathioprine‐induced vanishing bile duct syndrome: The value of early thiopurine metabolism assessment

Chouchana

Terris

Sogni

et al. 2023

Brit J Clinical Pharma

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About 15% to 28% of patients treated with thiopurines experienced adverse drug reactions, such as haematological and hepatic toxicities. Some of these related to the polymorphic activity of the thiopurine S‐methyltransferase (TPMT), the key detoxifying enzyme of thiopurine metabolism. We report here a case of thiopurine‐induced ductopenia with a comprehensive pharmacological analysis on thiopurine metabolism. A 34‐year‐old woman, with a medical history of severe systemic lupus erythematosus with recent introduction of azathioprine therapy, presented with mild fluctuating transaminase blood levels consistent with a hepatocellular pattern, which evolved to a cholestatic pattern over the next weeks. A blood thiopurine metabolite assay revealed low 6‐thioguanine nucleotides (6‐TGN) level and a dramatically increased 6‐methylmercaptopurine ribonucleotides (6‐MMPN) level, together with an unfavourable [6‐MMPN:6‐TGN] metabolite ratio and a high TPMT activity. After a total of about 6 months of thiopurine therapy, a transjugular liver biopsy revealed a ductopenia, and azathioprine discontinuation led to further clinical improvement. In line with previous reports from the literature, our case supports the fact that ductopenia is a rare adverse drug reaction of azathioprine. The mechanism of reaction is unknown but may involve high 6‐MMPN blood level, due to unusual thiopurine metabolism (switched metabolism). Early therapeutic drug monitoring with measurement of 6‐TGN and 6‐MMPN blood levels may help physicians to identify patients at risk of similar duct injury.

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“…There was no sign of haematological toxicity. A blood thiopurine metabolite assay was performed according to a previously described method, 7…”

Section: Case Reportmentioning

confidence: 99%

Section: Case Reportmentioning

confidence: 99%

Azathioprine‐induced vanishing bile duct syndrome: The value of early thiopurine metabolism assessment

Chouchana

Terris

Sogni

et al. 2023

Brit J Clinical Pharma

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“…After pre-treated by 240 uL Trichloroacetic Acid (400 g/L) with an ice bath for 30 min, 1.2 mL plasma was centrifuged at 14000rpm at 4°C for 20 min, and then supernatant was separated and ltered. HPLC-ultraviolet detection was used to measure plasma SAM and SAH concentrations according to a method presented elsewhere (Illamola et al, 2019). We quanti ed plasma SAM and SAH concentrations in accordance with the peak area with standard solutions of (0, 0.05078, 0.10156, 0.20312, 0.40625, 0.81250, 1.62500 and 3.25000 mg/L for SAM) and (0, 0.03125, 0.06250, 0.12500, 0.25000, 0.50000 and 1.00000 mg/L for SAH) to establish the retention time.…”

Section: Plasma Sahh Activitymentioning

confidence: 99%

Effects of H19/SAHH/DNMT1 on the oxidative DNA damage related to benzo[a]pyrene exposure

Li²,

Pan³

et al. 2022

Environ Sci Pollut Res

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The mechanisms that long noncoding RNA (lncRNA) H19 binding to S-adenosylhomocysteine hydrolase (SAHH) interacted with DNA methyltransferase 1 (DNMT1) and then regulated DNA damage caused by PAHs remain unclear. A total of 146 occupational workers in a Chinese coke-oven plant in 2014 were included in the nal analyses. We used high performance liquid chromatography mass spectrometry (HPLC-MS) equipped to detect urine biomarkers of PAHs exposure, including 2-hydroxynaphthalene (2-NAP), 2-hydroxy uorene (2-FLU), 9-hydroxyphenanthrene (9-PHE), 1-hydroxypyrene (1-OHP). The levels of SAM and SAH in plasma were detected by HPLC-ultraviolet. By constructing various BEAS-2B cell models exposed to 16 µM Benzo[a]pyrene (BaP) for 24 h, toxicological parameters re ecting distinct mechanisms were evaluated. We documented that urinary 1-hydroxypyrene (1-OHP) levels were positively associated with blood H19 RNA expression (OR: 1.51, 95% CI: 1.03 -2.19), but opposite to plasma SAHH activity (OR: 0.63, 95% CI: 0.41 -0.98) in coke oven workers. Moreover, by constructing various BEAS-2B cell models exposed to Benzo[a]pyrene (BaP), we investigated that H19 binding to SAHH exaggerated DNMT1 expressions and activity. Suppression of H19 enhanced the interaction of SAHH and DNMT1 in BaPtreated cells, decreased OGG1 methylation, reduced oxidative DNA damage and lessened S phase arrest. However, SAHH or DNMT1 single knockdown and SAHH/DNMT1 double knockdown showed the opposite trend. A H19/SAHH/DNMT1 axis was involved in OGG1 methylation, oxidative DNA damage and cell cycle arrest by carcinogen BaP.

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“…After pre-treated by 240 uL Trichloroacetic Acid (400 g/L) with an ice bath for 30 min, 1.2 mL plasma was centrifuged at 14000rpm at 4°C for 20 min, and then supernatant was separated and ltered. HPLCultraviolet detection was used to measure plasma SAM and SAH concentrations according to a method presented elsewhere (35). We quanti ed plasma SAM and SAH concentrations in accordance with the peak area with standard solutions of (0, 0.05078, 0.10156, 0.20312, 0.40625, 0.81250, 1.62500 and 3.25000 mg/L for SAM) and (0, 0.03125, 0.06250, 0.12500, 0.25000, 0.50000 and 1.00000 mg/L for SAH) to establish the retention time.…”

Section: Plasma Sahh Activitymentioning

confidence: 99%

Effects of H19/SAHH/DNMT1 on the oxidative DNA damage related to Benzo[a]pyrene exposure

Pan

et al. 2021

Preprint

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Background: The mechanisms that long noncoding RNA (lncRNA) H19 binding to S-adenosylhomocysteine hydrolase (SAHH) interacted with DNA methyltransferase 1 (DNMT1) and then regulated DNA damage caused by PAHs remain unclear. Results: We documented that urinary 1-hydroxypyrene (1-OHP) levels were positively associated with blood H19 RNA expression (OR: 1.51, 95% CI: 1.03 - 2.19), but opposite to plasma SAHH activity (OR: 0.63, 95% CI: 0.41 - 0.98) in coke oven workers. Moreover, by constructing various BEAS-2B cell models exposed to Benzo[a]pyrene (BaP), we investigated that H19 binding to SAHH exaggerated DNMT1 expressions and activity. Suppression of H19 enhanced the interaction of SAHH and DNMT1 in BaP-treated cells, decreased OGG1 methylation, reduced oxidative DNA damage and lessened S phase arrest. However, SAHH or DNMT1 single knockdown and SAHH/DNMT1 double knockdown showed the opposite trend. Conclusions: A H19/SAHH/DNMT1 axis was involved in OGG1 methylation, oxidative DNA damage and cell cycle arrest by carcinogen BaP.

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Development and validation of a UPLC-UV method for the quantification of thiopurine methyltransferase enzyme activity in human erythrocytes

Cited by 6 publications

References 27 publications

Azathioprine‐induced vanishing bile duct syndrome: The value of early thiopurine metabolism assessment

Azathioprine‐induced vanishing bile duct syndrome: The value of early thiopurine metabolism assessment

Effects of H19/SAHH/DNMT1 on the oxidative DNA damage related to benzo[a]pyrene exposure

Effects of H19/SAHH/DNMT1 on the oxidative DNA damage related to Benzo[a]pyrene exposure

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