Genomic and Proteomic Profiling of Responses to Toxic Metals in Human Lung Cells

Andrew, Angeline S.; Warren, Amy J.; Barchowsky, Aaron; Temple, Kaili A.; Klei, Linda R.; Soucy, Nicole V.; O’Hara, Kimberley A.; Hamilton, Joshua W.

doi:10.1289/txg.6249

Cited by 17 publications

(22 citation statements)

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“…Prior studies have also reported genome-wide gene expression changes in BEAS-2B cells exposed to arsenite. Andrew et al (2003) measured gene expression in confluent cultures of BEAS-2B grown in defined LHC-9 medium, using matrix (fibronectin/bovine collagen/bovine albumin)-coated flasks. That study used a targeted analysis of 1200 genes in cells that had been exposed to 5 or 50 uM arsenite for 4 hours.…”

Section: Discussionmentioning

confidence: 99%

Culture conditions profoundly impact phenotype in BEAS‐2B, a human pulmonary epithelial model

Zhao

Klimecki

2014

J of Applied Toxicology

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BEAS-2B, an immortalized, human lung epithelial cell line, has been used to model pulmonary epithelial function for over 30 years. The BEAS-2B phenotype can be modulated by culture conditions that include the presence or absence of fetal bovine serum (FBS). The popularity of BEAS-2B as a model of arsenic toxicology, and the common use of BEAS-2B cultured both with and without FBS, led us to investigate the impact of FBS on BEAS-2B in the context of arsenic toxicology. Comparison of genome-wide gene expression in BEAS-2B cultured with or without FBS revealed altered expression in several biological pathways, including those related to carcinogenesis and energy metabolism. Real-time measurements of oxygen consumption and glycolysis in BEAS-2B demonstrated that FBS culture conditions were associated with a 1.4-fold increase in total glycolytic capacity, a 1.9-fold increase in basal respiration, a 2.0-fold increase in oxygen consumed for ATP production, and a 2.8-fold increase in maximal respiration, compared to BEAS-2B cultured without FBS. Comparisons of the transcriptome changes in BEAS-2B resulting from FBS exposure to the transcriptome changes resulting from exposure to 1 μM sodium arsenite revealed that mRNA levels of 43% of the arsenite-modulated genes were also modulated by FBS. Cytotoxicity studies revealed that BEAS-2B cells exposed to 5% FBS for 8 weeks were almost 5 times more sensitive to arsenite cytotoxicity than non-FBS-exposed BEAS-2B cells. Phenotype changes induced in BEAS-2B by FBS suggest that culture conditions should be carefully considered when using BEAS-2B as an experimental model of arsenic toxicity.

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Section: Discussionmentioning

confidence: 99%

Culture conditions profoundly impact phenotype in BEAS‐2B, a human pulmonary epithelial model

Zhao

Klimecki

2014

J of Applied Toxicology

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“…Like iAs +3 , iAs +5 is complexed with oxygen (as arsenate), but arsenate is charged (and isostructural with phosphate and sulfate). Thus, arsenate may not freely diffuse into the cell, and it enters via the so-called non-specific anion transporter and other phosphate transport systems (Andrew et al, 2003), in competition with the high concentration of phosphate in serum (Druwe and Vaillancourt, 2010). Thus, As +3 is more readily taken up into the cell, in comparison with As +5 (Druwe and Vaillancourt, 2010;Zangi and Filella, 2012).…”

Section: Introductionmentioning

confidence: 99%

Monomethylated trivalent arsenic species disrupt steroid receptor interactions with their DNA response elements at non‐cytotoxic cellular concentrations

Gosse

Taylor

Jackson

et al. 2013

J of Applied Toxicology

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Arsenic (As) is considered a top environmental chemical of human health because it has been linked to adverse health effects including cancer, diabetes, cardiovascular disease, and reproductive and developmental problems. In several cell culture and animal models, As acts as an endocrine disruptor, which may underlie many of its health effects. Previous work showed that steroid receptor (SR)-driven gene expression is disrupted in cells treated with inorganic As (arsenite, iAs+3). In those studies, low iAs+3 concentrations (0.1–0.7 μM) stimulated hormone-inducible transcription, whereas somewhat higher but still non-cytotoxic levels (1–3 μM) inhibited transcription. This investigation focuses on the mechanisms underlying these inhibitory effects and evaluates the role of methylated trivalent As metabolites on SR function. Recent evidence suggests that, compared with iAs, methylated forms may have distinct biochemical effects. Here, fluorescence polarization (FP) experiments utilizing purified, hormone-bound human glucocorticoid (GR) and progesterone receptor (PR) have demonstrated that neither inorganic (iAs+3) nor dimethylated (DMA+3) species of trivalent As affect receptor interactions with glucocorticoid DNA response elements (GREs). However, monomethylated forms (monomethylarsenite, MMA+3 and monomethylarsonic diglutathione, MADG) strongly inhibit GR-GRE and PR-GRE binding. Additionally, speciation studies of iAs+3-treated H4IIE rat hepatoma cells show that, under treatment conditions that cause inhibition of hormone-inducible gene transcription, the intracellular concentration of MADG is sufficient to inhibit GR-GRE and PR-GRE interactions in vivo. These results indicate that arsenic’s inhibitory endocrine disruption effects are probably caused in part by methylated metabolites’ disruption of SR ability to bind DNA response elements that are crucial to hormone-driven gene transcription.

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“…However, epidemiological data suggest that certain cumulative threshold doses need to be reached for carcinogenic effect over a long period of time, whereas we use LDA only for a very short period of time and have not observed any secondary malignancy with LDA in our mouse model (Ganapathy et al, 2014a;Snow et al, 2005). It has been well documented in vitro that arsenic has different biological effects and expresses different sets of genes depending on the dose, supporting out strategy (Andrew et al, 2003). We have taken this strategy from our preclinical studies to the current clinical trial.…”

Section: Introductionmentioning

confidence: 53%

p53‐Based strategy to reduce hematological toxicity of chemotherapy: A proof of principle study

Michalek

Elledge

et al. 2015

Molecular Oncology

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P53 activation is a primary mechanism underlying pathological responses to DNA-damaging agents such as chemotherapy and radiotherapy. Our recent animal studies showed that low dose arsenic (LDA)-induced transient p53 inhibition selectively protected normal tissues from chemotherapy-induced toxicity. Study objectives were to: 1) define the lowest safe dose of arsenic trioxide that transiently blocks p53 activation in patients and 2) assess the potential of LDA to decrease hematological toxicity from chemotherapy. Patients scheduled to receive minimum 4 cycles of myelosuppressive chemotherapy were eligible. For objective 1, dose escalation of LDA started at 0.005mg/kg/day for 3 days. This dose satisfied objective 1 and was administered before chemotherapy cycles 2, 4 and 6 for objective 2. P53 level in peripheral lymphocytes was measured on day 1 of each cycle by ELISA assay. Chemotherapy cycles 1, 3, and 5 served as the baseline for the subsequent cycles of 2, 4 and 6 respectively. If p53 level for the subsequent cycle was lower (or higher) than the baseline cycle, p53 was defined as “suppressed” (or “activated”) for the pair of cycles. Repeated measures linear models of CBC in terms of day, cycle, p53 activity and interaction terms were used. Twenty-six patients treated with 3 week cycle regimens form the base of analyses. The mean white blood cell, hemoglobin and absolute neutrophil counts were significantly higher in the “suppressed” relative to the “activated” group. These data support the proof of principle that suppression of p53 could lead to protection of bone marrow in patients receiving chemotherapy.

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Genomic and Proteomic Profiling of Responses to Toxic Metals in Human Lung Cells

Cited by 17 publications

References 0 publications

Culture conditions profoundly impact phenotype in BEAS‐2B, a human pulmonary epithelial model

Culture conditions profoundly impact phenotype in BEAS‐2B, a human pulmonary epithelial model

Monomethylated trivalent arsenic species disrupt steroid receptor interactions with their DNA response elements at non‐cytotoxic cellular concentrations

p53‐Based strategy to reduce hematological toxicity of chemotherapy: A proof of principle study

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