Purpose
To bring together researchers and other stakeholders with an interest in nucleic acid (DNA, RNA) and protein adductomics to promote collaboration and establish guidelines and best practices. The consortium will focus on becoming a reference for anyone interested in adductomics and ultimately it will promote robust analytical approaches to advance the field. This effort will be promoted through the creation of a website to be used as a common platform to share information and resources, the organization of events at scientific meetings focusing on the presentation and discussion of adductomic research and finally the facilitation of inter-laboratory initiatives to promote standardization procedures. EMGS and the Division of Chemical Toxicology of the American Chemical Society could be organizations interested in promoting the initiatives of the consortium. The EMGS has already funded a small pilot proposal to support the initiation of these efforts.
Aims
DNA/RNA:
4. To compare the results processed by the existing post-data software (there are at least four software packages for assessing adductomics data reported in the literature to date).
5. To explore inter- and intra-lab and platform variability.
6. To develop a website to provide a comprehensive resource for adductomics researchers and other stakeholders.
To bring together researchers and other stakeholders with an interest in nucleic acid (DNA, RNA) and protein adductomics to promote collaboration and establish guidelines and best practices. The consortium will focus on becoming a reference for anyone interested in adductomics and ultimately it will promote robust analytical approaches to advance the field. This effort will be promoted through the creation of a website to be used as a common platform to share information and resources, the organization of events at scientific meetings focusing on the presentation and discussion of adductomic research and finally the facilitation of inter-laboratory initiatives to promote standardization procedures. EMGS and the Division of Chemical Toxicology of the American Chemical Society could be organizations interested in promoting the initiatives of the consortium. The EMGS has already funded a small pilot proposal to support the initiation of these efforts.
Aims
- To develop new, or expand existing, adduct database platforms, for both DNA/RNA and protein adducts. These platforms should be freely accessible, with IAC members able to add or update the information anytime as needed.
- To share resources and develop and agree on best practices for sample preparation methods and LC-MS measurement conditions.
DNA/RNA:
- purification process (e.g., manual SPE or centrifugal device, if needed)
- enzymatic hydrolysis (e.g., optimal cocktail and conditions for enzymes),
- nucleic acids extraction (e.g., identifying the optimal conditions for each type of adduct, and/or the broadest range of adducts),
- Protein extraction and purification
- Protein digestion (e.g., pressure cycling technology)
- Conditions for optimal peptide stability
- Minimal acceptable mass resolution.
- Acceptable m/z tolerances.
- Relative retention time considerations (e.g., use of normalized retention times - https://doi.org/10.1021/acs.analchem.8b04660).
- Use of intensity for semi-quantification of adduct levels.
- Use of community-established internal standards (see 3, below).
- use of HRMS (untargeted adductomics) versus QQQ-MS (targeted adductomics)
- acceptable m/z tolerances
- relative retention time considerations
- use of housekeeping peptides for normalization
- use of internal standards
- adduct discovery software
4. To compare the results processed by the existing post-data software (there are at least four software packages for assessing adductomics data reported in the literature to date).
5. To explore inter- and intra-lab and platform variability.
6. To develop a website to provide a comprehensive resource for adductomics researchers and other stakeholders.
Next Steps:
Next/Parallel steps:
- To advocate for the field of adductomics, and raise awareness with the broader research and industry communities (e.g., funding agencies, mass spectrometry manufacturers and data analysis software developers).
- To explore, and identify sources of variability in the adductome (e.g., methodological, diurnal, intra-, and individual, and depending on the samples/tissues considered
- To accelerate the interplay between mass spectrometry and single molecule DNA sequencing (e.g., Oxford nanopore) for site-specific detection of chemically characterized DNA adducts.
- To explore adducts on additional proteins (e.g., albumin, hemoglobin, others) and additional amino acid targets (e.g., Cys, Lys, global approaches).
- To encourage and support junior faculty and students to join the adductomics field, and become the next generation of adduct and adductomics researchers.
Mission Statement
"Bringing together researchers with an interest in nucleic acid (DNA, RNA) and protein adductomics to promote collaboration and establish guidelines for best practices. "
