Assessment of systemic genetic damage in pediatric inflammatory bowel disease

Baig, Ayesha; Avlasevich, Svetlana L.; Torous, Dorothea K.; Bemis, Jeffrey C.; Saubermann, Lawrence J.; Lovell, David P.; MacGregor, James T.; Dertinger, Stephen D.

doi:10.1002/em.22403

Cited by 8 publications

(11 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…was 3.2 ± 2.7 and 2.3 ± 1.6 ( p = .4712) for the untreated and treated groups, respectively. Furthermore, the range of low mutant cell frequencies are consistent with previous reports that studied healthy pediatric subjects (Torous et al, 2020; Baig et al, 2020).…”

Section: Resultssupporting

confidence: 91%

“…Within 24 h of collection, aliquots of Wilmot Cancer Center‐derived blood samples were fixed with ultra‐cold absolute methanol by Litron Laboratories personnel for the MN‐RET assay as previously described (Torous et al, 2020; Baig et al, 2020). After at least 3 days in ultra‐cold storage, samples were moved into In Vivo MicroFlow® Kit's Long‐Term Storage Solution and were stored between −75 and −85°C until the day of analysis.…”

Section: Methodsmentioning

confidence: 99%

“…Within 24 h of collection, aliquots of Wilmot Cancer Center‐derived blood samples were frozen with In Vivo MutaFlow® Kit's Blood Freezing Solution for the MUT RET assay by Litron personnel as previously described (Torous et al, 2020; Baig et al, 2020). The samples were stored between −75 and−85°C until the day of analysis.…”

Section: Methodsmentioning

confidence: 99%

“…The methods for preparing blood samples for MN‐RET scoring via flow cytometric analysis have been described in detail (Torous et al, 2020; Baig et al, 2020). The method is based on the enrichment of rare peripheral blood CD71‐positive reticulocytes through immunomagnetic separation.…”

Section: Methodsmentioning

confidence: 99%

See 3 more Smart Citations

Lack of hydroxyurea‐associated mutagenesis in pediatric sickle cell disease patients

Torous

Avlasevich

Bemis

et al. 2023

Environ and Mol Mutagen

Self Cite

View full text Add to dashboard Cite

Hydroxyurea is approved for treating children and adults with sickle cell anemia (SCA). Despite its proven efficacy, concerns remain about its mutagenic and carcinogenic potential that hamper its widespread use. Cell culture-and animal-based investigations indicate that hydroxyurea's genotoxic effects are due to indirect clastogenicity in select cell types when high dose and time thresholds are exceeded (reviewed by Ware & Dertinger, 2021). The current study extends these preclinical observations to pediatric patients receiving hydroxyurea for treatment of SCA. First, proof-of-principle experiments with testicular cancer patients exposed to a cisplatinbased regimen validated the ability of flow cytometric blood-based micronucleated reticulocyte (MN-RET) and PIG-A mutant reticulocyte (MUT RET) assays to detect clastogenicity and gene mutations, respectively. Second, these biomarkers were measured in a cross-sectional study with 26 SCA patients receiving hydroxyurea and 13 SCA patients without exposure. Finally, a prospective study was conducted with 10 SCA patients using pretreatment blood samples and after 6 or 12 months of therapy. Cancer patients exposed to cisplatin exhibited increased MN-RET within days of exposure, while the MUT RET endpoint required more time to reach maximal levels.In SCA patients, hydroxyurea induced MN-RET in both the cross-sectional and prospective studies. However, no evidence of PIG-A gene mutation was found in hydroxyurea-treated children, despite the fact that the two assays use the same rapidly-dividing, highly-exposed cell type. Collectively, these results reinforce the complementary nature of MN-RET and MUT RET biomarkers, and indicate that hydroxyurea can be clastogenic but was not mutagenic in young patients with SCA.

show abstract

Section: Resultssupporting

confidence: 91%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Lack of hydroxyurea‐associated mutagenesis in pediatric sickle cell disease patients

Torous

Avlasevich

Bemis

et al. 2023

Environ and Mol Mutagen

Self Cite

View full text Add to dashboard Cite

show abstract

“…Two publications have assessed PIG‐A mutant cells in IBD patients. First, Baig et al (2020) found no significant increase in mutant reticulocytes in pediatric IBD patients ( n = 64) compared to controls ( n = 55) although 10 patients with higher micronucleus frequencies all had established disease. Second, Cao, Wang, Liu, et al (2020) found significantly elevated erythrocyte mutant cell number in adult Chinese IBD patients ( n = 36) compared to age‐ and sex‐matched controls ( n = 36), although these patients were all undergoing azathioprine treatment.…”

Section: The Pig‐a Mutation Test In Diseasementioning

confidence: 97%

The PIG‐A gene mutation assay in human biomonitoring and disease

Lawrence,

Munn,

Naser

et al. 2023

Environ and Mol Mutagen

View full text Add to dashboard Cite

The blood cell phosphatidylinositol glycan class A (PIG‐A) gene mutation assay has been extensively researched in rodents for in vivo mutagenicity testing and is now being investigated in humans. The PIG‐A gene is involved in glycosyl phosphatidylinositol (GPI)‐anchor biosynthesis. A single mutation in this X‐linked gene can lead to loss of membrane‐bound GPI anchors, which can be enumerated via corresponding GPI‐anchored proteins (e.g., CD55) using flow cytometry. The studies published to date by different research groups demonstrate a remarkable consistency in PIG‐A mutant frequencies. Moreover, with the low background level of mutant erythrocytes in healthy subjects (2.9–5.56 × 10−6 mutants), induction of mutation post genotoxic exposure can be detected. Cigarette smoking, radiotherapy, and occupational exposures, including lead, have been shown to increase mutant levels. Future applications of this test include identifying new harmful agents and establishing new exposure limits. This mutational monitoring approach may also identify individuals at higher risk of cancer development. In addition, identifying protective agents that could mitigate these effects may reduce baseline somatic mutation levels and such behaviors can be encouraged. Further technological progress is required including establishing underlying mechanisms of GPI anchor loss, protocol standardization, and the development of cryopreservation methods to improve GPI‐anchor stability over time. If successful, this assay has the potential be widely employed, for example, in rural and low‐income countries. Here, we review the current literature on PIG‐A mutation in humans and discuss the potential role of this assay in human biomonitoring and disease detection.

show abstract