Dataset Information

FIRST ATTEMPT TO COUPLE PROTEOMIC WITH AhR REPORTER GENE BIOASSAY IN SOIL POLLUTION MONITORING AND ASSESSMENT

ABSTRACT: A topsoil sample obtained from a highly industrialized area was tested on CALUX® cell line and exposed cells processed with proteomic and bioinformatics analyses. GC-MS/MS analysis of topsoil extracts was also performed to confirm PCDD, PCDF and dl-PCBs presence. CALUX® and GC-MS/MS data confirmed the presence of PCDD/Fs and dl-PCBs in the topsoil extract. Proteomic analysis of the cells exposed to the soil extracts identified 43 differential proteins. Enrichment analysis highlights biological processes such as cellular response to chemical stimulus, to stress, response to inorganic substance, regulation of translation, regulation of apoptotic process as well as response to organonitrogen compounds in light of particular drugs and compounds, extrapolated by bioinformatics and linked to the identified proteins modifications. These results confirm the complex epidemiological situation occurring among Taranto inhabitants and underline the need to further investigate the presence and sources of inferred chemicals in Taranto soils. The combined analysis reveals a more complex scenarios of chemicals able to affect cellular pathways and leading to toxicities rather than those identified by the bioassay. The approach to couple proteomic to CALUX® is a promising tool and deserves further investigation to be applied in soil pollution monitoring and risk assessment.

INSTRUMENT(S): autoflex

ORGANISM(S): Rattus Norvegicus (rat)

TISSUE(S): Cell Culture

SUBMITTER: Enxhi Shaba

LAB HEAD: Prof. Luca Bini

PROVIDER: PXD027074 | Pride | 2022-02-17

REPOSITORIES: Pride

ACCESS DATA

Publications

First Attempt to Couple Proteomics with the <i>AhR</i> Reporter Gene Bioassay in Soil Pollution Monitoring and Assessment.

Landi Claudia C Liberatori Giulia G Cotugno Pietro P Sturba Lucrezia L Vannuccini Maria Luisa ML Massari Federica F Miniero Daniela Valeria DV Tursi Angelo A Shaba Enxhi E Behnisch Peter A PA Carleo Alfonso A Di Giuseppe Fabrizio F Angelucci Stefania S Bini Luca L Corsi Ilaria I

Toxics 20211229 1

A topsoil sample obtained from a highly industrialized area (Taranto, Italy) was tested on the DR-CALUX<sup>®</sup> cell line and the exposed cells processed with proteomic and bioinformatics analyses. The presence of polyhalogenated compounds in the topsoil extracts was confirmed by GC-MS/MS analysis. Proteomic analysis of the cells exposed to the topsoil extracts identified 43 differential proteins. Enrichment analysis highlighted biological processes, such as the cellular response to a chemic ...[more]

PMID: 35051051

Similar Datasets

Project description:Introduction: Higher-throughput mode-of-action-based assays provide a valuable approach to expedite chemical evaluation for human health risk assessment. In this study, we combined the high-throughput alkaline CometChip® assay with the TGx-DDI transcriptomic biomarker (DDI = DNA damage-inducing) using high-throughput TempO-Seq® as an integrated genotoxicity testing approach. Objective: To determine if the high-throughput CometChip® assay and TGx-DDI biomarker analysis can be combined using human HepaRG™ cells to provide an efficient next-generation genotoxicity screening approach to identify DNA damage-inducing (DDI) chemicals. Methods: Metabolically competent human HepaRG™ cell cultures were exposed to increasing concentrations of 12 chemicals (9 DDI and 3 non-DDI) using a repeat exposure design (0 hr, 24 hr, 48 hr) in 96-well plates to measure and classify chemicals based on their ability to damage DNA. After a 4 hr recovery period, exposed cells were used to perform the Trevigen CometChip assay to assess DNA damage or used for TempO-Seq® S1500+ Targeted Transcriptome Sequencing for TGx-DDI classification purposes. Bioinformatics and statistical tools were used to classify chemicals as DDI or non-DDI using three separate analyses. Benchmark concentration analysis was also conducted for both assays for the purposes of potency ranking. Results: In combination, the CometChip® assay and the TGx-DDI were 100% accurate in identifying chemicals that induce DNA damage. Quantitative benchmark concentration (BMC) modeling was applied to evaluate chemical potencies for both assays. The BMCs for the CometChip® assay and the TGx-DDI for all 12 chemicals were highly concordant (within four-fold) and resulted in identical potency rankings. Conclusions: These results suggest that these two assays can be integrated for efficient, high-throughput identification of DNA damage in HepaRG™ cells. This study provides evidence for the complementarity of the high-throughput CometChip® assay with TGx-DDI biomarker analysis. Integration of these two tests provides an effective and high-throughput approach to identify DDI agents. This work is a first step in accomplishing a more integrated genotoxicity testing strategy to derive mechanistic information to better inform human health risk assessment in a high-throughput manner.

Dataset Information

FIRST ATTEMPT TO COUPLE PROTEOMIC WITH AhR REPORTER GENE BIOASSAY IN SOIL POLLUTION MONITORING AND ASSESSMENT

Publications

First Attempt to Couple Proteomics with the <i>AhR</i> Reporter Gene Bioassay in Soil Pollution Monitoring and Assessment.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure