Use of the γ-H2AX assay to monitor DNA damage and repair in translational cancer research

Ivashkevich, Alesia; Redon, Christophe E.; Nakamura, Asako; Martin, Roger F.; Martin, Olga A.

doi:10.1016/j.canlet.2011.12.025

Cited by 375 publications

(292 citation statements)

References 138 publications

Supporting

Mentioning

275

Contrasting

Order By: Relevance

“…When cells are exposed to ionizing radiation or DNA-damaging chemotherapeutic agents, DSBs are generated that rapidly result in the phosphorylation of histone H2A variant H2AX. Because phosphorylation of H2AX at Ser 139 (γ-H2AX) correlates well with each DSB, it is the most sensitive marker that can be used to examine the DNA damage produced and the subsequent repair of the DNA lesion (30,31). We used immunofluorescence staining γ-H2AX to detect DSB in the event of H.japonicum infection.…”

Section: Helicobacter Japonicum Infection Caused Dna Doublestrand Brementioning

confidence: 99%

NovelHelicobacterspeciesH.japonicumisolated from laboratory mice from Japan induces typhlocolitis and lower bowel carcinoma in C57BL/129 IL10^−/−mice

Shen¹,

Feng²,

Muthupalani³

et al. 2016

CARCIN

View full text Add to dashboard Cite

A novel Helicobacter species Helicobacter japonicum was isolated from the stomach and intestines of clinically normal mice received from three institutes from Japan. The novel Helicobacter sp. was microaerobic, grew at 37°C and 42°C, was catalase and oxidase positive, but urease negative. It is most closely related to the 16S rRNA gene of H.muridarum (98.6%); to the 23S rRNA gene of H.hepaticus (97.9%); to the hsp60 gene of H.typhlonius (87%). The novel Helicobacter sp. has in vitro cytolethal distending toxin (CDT) activity; its cdtB gene sequence has 83.8% identity with that of H.hepaticus. The whole genome sequence of H.japonicum MIT 01-6451 has a 2.06-Mb genome length with a 37.5% G + C content. When the organism was inoculated into C57BL/129 IL10 −/− mice, it was cultured from the stomach, colon and cecum of infected mice at 6 and 10 weeks post-infection. The cecum had the highest H.japonicum colonization levels by quantitative PCR. The histopathology of the lower bowel was characterized by moderate to severe inflammation, mild edema, epithelial defects, mild to severe hyperplasia, dysplasia and carcinoma. Inflammatory cytokines IFNγ, TNFα and IL17a, as well as iNOS were significantly upregulated in the cecal tissue of infected mice. These results demonstrate that the novel H.japonicum can induce inflammatory bowel disease and carcinoma in IL10 −/− mice and highlights the importance of identifying novel Helicobacter spp. especially when they are introduced from outside mouse colonies from different geographic locations.

show abstract

Section: Helicobacter Japonicum Infection Caused Dna Doublestrand Brementioning

confidence: 99%

NovelHelicobacterspeciesH.japonicumisolated from laboratory mice from Japan induces typhlocolitis and lower bowel carcinoma in C57BL/129 IL10^−/−mice

Shen¹,

Feng²,

Muthupalani³

et al. 2016

CARCIN

View full text Add to dashboard Cite

show abstract

“…In our laboratories, we used the g histone 2A isoform X (H2AX) assay 32,33 for detecting DNA DSBs in irradiated primary fibroblasts established from skin biopsy samples from SCID and CID patients. g-H2AX is a de novo protein formed in response to DSB formation.…”

mentioning

confidence: 99%

“…These properties make the g-H2AX an attractive screening biomarker in translational research, for the assessment of clinical biodosimetry of diagnostic and therapeutic radiation and DNA-damaging chemotherapy. 33,42,43 We and others have demonstrated that the number and kinetics of decline of radiation-induced foci (surrogate of DSB repair) are a measure of the cellular RS in SCID and CID applications and other settings. 6e8, 40,44,45 Recently, we presented several analytical tools for improving the statistical and computational approaches to applying the assay for differential diagnostics in RS and non-RS biodosimetry in tissues and in the primary fibroblast skin culture model.…”

mentioning

confidence: 99%

Evaluation of Severe Combined Immunodeficiency and Combined Immunodeficiency Pediatric Patients on the Basis of Cellular Radiosensitivity

Lobachevsky

Woodbine

Hsiao

et al. 2015

The Journal of Molecular Diagnostics

Self Cite

View full text Add to dashboard Cite

Pediatric patients with severe or nonsevere combined immunodeficiency have increased susceptibility to severe, life-threatening infections and, without hematopoietic stem cell transplantation, may fail to thrive. A subset of these patients have the radiosensitive (RS) phenotype, which may necessitate conditioning before hematopoietic stem cell transplantation, and this conditioning includes radiomimetic drugs, which may significantly affect treatment response. To provide statistical criteria for classifying cellular response to ionizing radiation as the measure of functional RS screening, we analyzed the repair capacity and survival of ex vivo irradiated primary skin fibroblasts from five dysmorphic and/or developmentally delayed pediatric patients with severe combined immunodeficiency and combined immunodeficiency. We developed a mathematical framework for the analysis of g histone 2A isoform X foci kinetics to quantitate DNA-repair capacity, thus establishing crucial criteria for identifying RS. The results, presented in a diagram showing each patient as a point in a 2D RS map, were in agreement with findings from the assessment of cellular RS by clonogenic survival and from the genetic analysis of factors involved in the nonhomologous end-joining repair pathway. We provide recommendations for incorporating into clinical practice the functional assays and genetic analysis used for establishing RS status before conditioning. This knowledge would enable the selection of the most appropriate treatment regimen, reducing the risk for severe therapy-related adverse effects. Severe combined immunodeficiency (SCID) and combined immunodeficiency (CID) are rare genetic disorders. The incidence of SCID in Australia is 1 in 69,000 live births, 1 comparable to the reported incidence of 1 in 100,000 births worldwide. CID patients have increased susceptibility to invasive and opportunistic bacterial, viral, and fungal infections due to poor T-lymphocyte production and/or function, in addition to failure of B lymphocytes to generate functional antibodies. SCID is the extreme form of CID. Patients with this condition often present in the first year of life, with severe life-threatening infections, and consequently failure to thrive, requiring prompt intervention. SCID without treatment is usually fatal within the first year of life. 2 Both SCID and CID are genetically diverse syndromes, and over 50 molecular defects resulting in these syndromes have been described. 3 Approximately 30% of SCID patients have defects in V(D)J recombination (antigen receptor

show abstract

“…A microscopy‐based γ H2AX assay allows to measure γ H2AX foci in different cell types from a same tissue simultaneously. For all these reasons, γ H2AX has been widely used in the last decade to monitor both irradiation and cancer drug responses in patients 17. While pharmacodynamics should be preferentially performed in tumor biopsies, tumor collection may be problematic and/or unsafe for patients.…”

Section: Resultsmentioning

confidence: 99%

Evaluation of surrogate tissues as indicators of drug activity in a melanoma skin model

et al. 2016

Self Cite

View full text Add to dashboard Cite

The development of novel cancer treatments is a challenging task, partly because results from model systems often fail to predict drug efficacy in humans, and also tumors are often inaccessible for biochemical analysis, preventing effective monitoring of drug activity in vivo. Utilizing a model system, we evaluated the use of drug‐induced DNA damage in surrogate tissues as indicators of drug efficacy. Samples of a commercially available melanoma skin model (Mattek MLNM‐FT‐A375) containing keratinocyte and fibroblast layers with melanoma nodules were subjected to various chemotherapeutic regimens for one, four, or eight days. At these times they were analyzed for DNA double‐stranded breaks (γH2AX foci) and apoptosis (TUNEL). A wide range of drug responses in both tumor and normal tissues were observed and cataloged. For the melanoma, the most common drug response was apoptosis. The basal keratinocyte layer, which was the most reliable indicator of drug response in the melanoma skin model, responded with γH2AX foci formation that was abrupt and transient. The relationships between tumor and surrogate tissue drug responses are complex, indicating that while surrogate tissue drug responses may be useful clinical tools, careful control of variables such as the timing of sampling may be important in interpreting the results.

show abstract

Use of the γ-H2AX assay to monitor DNA damage and repair in translational cancer research

Cited by 375 publications

References 138 publications

NovelHelicobacterspeciesH.japonicumisolated from laboratory mice from Japan induces typhlocolitis and lower bowel carcinoma in C57BL/129 IL10^−/−mice

NovelHelicobacterspeciesH.japonicumisolated from laboratory mice from Japan induces typhlocolitis and lower bowel carcinoma in C57BL/129 IL10^−/−mice

Evaluation of Severe Combined Immunodeficiency and Combined Immunodeficiency Pediatric Patients on the Basis of Cellular Radiosensitivity

Evaluation of surrogate tissues as indicators of drug activity in a melanoma skin model

Contact Info

Product

Resources

About

Use of the γ-H2AX assay to monitor DNA damage and repair in translational cancer research

Cited by 375 publications

References 138 publications

NovelHelicobacterspeciesH.japonicumisolated from laboratory mice from Japan induces typhlocolitis and lower bowel carcinoma in C57BL/129 IL10−/−mice

NovelHelicobacterspeciesH.japonicumisolated from laboratory mice from Japan induces typhlocolitis and lower bowel carcinoma in C57BL/129 IL10−/−mice

Evaluation of Severe Combined Immunodeficiency and Combined Immunodeficiency Pediatric Patients on the Basis of Cellular Radiosensitivity

Evaluation of surrogate tissues as indicators of drug activity in a melanoma skin model

Contact Info

Product

Resources

About

NovelHelicobacterspeciesH.japonicumisolated from laboratory mice from Japan induces typhlocolitis and lower bowel carcinoma in C57BL/129 IL10^−/−mice

NovelHelicobacterspeciesH.japonicumisolated from laboratory mice from Japan induces typhlocolitis and lower bowel carcinoma in C57BL/129 IL10^−/−mice