Tsang, K.K. & A.G. McArthur. 2017. Encoding the efflux pump phenomena. Oral presentation at the Second American Society for Microbiology Meeting on Rapid Applied Microbial Next-Generation Sequencing and Bioinformatics Pipelines, Washington, D.C.
Background: Efflux pumps are a major mechanism for intrinsic and acquired resistance to our current antibiotic armamentarium. Efflux mechanisms interplay synergistically with other resistance mechanisms, including drug permeability, degradation and inactivation, to strengthen pathogen antimicrobial resistance levels. Despite their clinical relevance, there is no resource that seeks to understand and predict the contribution of efflux pumps in antimicrobial resistance from genome sequence. This has resulted in limited prediction of the full potential of all resistance determinants in a bacterial cell.
Methods: The Comprehensive Antibiotic Resistance Database (CARD, https://card.mcmaster.ca/) and Resistance Gene Identifier (RGI) were optimized for E. coli and P. aeruginosa efflux pump detection through extensive curation and algorithmic development. Literature was mined and analyzed to curate all published information on E. coli and P. aeruginosa efflux pumps into CARD. Algorithmic development of RGI involved creating bioinformatics detection models and refining their parameters. The Efflux Pump Identifier (EPI) was developed to predict efflux pumps and antimicrobial resistance based on RGI results generated using CARD and tested using genome sequences of characterized, clinical multi-drug resistant E. coli and P. aeruginosa isolates.
Results: The Efflux Pump Identifier (EPI) analyzed 124 E. coli and 94 P. aeruginosa clinical multi-drug resistant samples to predict efflux pumps and their complex regulatory networks under three paradigms: 1) Perfect, 2) Partial, and 3) Putative. The Perfect paradigm identifies perfect matches to known efflux pumps curated into CARD. The Partial algorithm detects efflux pumps where at least one or more components of the efflux pump is not a perfect match to an efflux pump component in CARD, but likely a functional homolog. Lastly, the Putative algorithm discovers potential efflux pumps where all components are not perfect matches to previously curated components in CARD.
Conclusions: The development of the Efflux Pump Identifier (EPI) devotes effort to an area in antimicrobial resistance where insufficient attention has been paid in the past. This is a step towards answering the long-standing question in the efflux pump phenomena; is the detected efflux pump genotype being expressed to present a specific phenotype? Using the Efflux Pump Identifier (EPI) in tandem with the existing repertoire of detection tools for dedicated and mutational resistance determinants leads to the complete prediction of antibiogram from genome sequence.
Building upon her successful Biochem 3A03 project, Tammy Lau is staying in the lab for 2017-2018 as part of her Biochem 4T15 Research Thesis. Tammy’s research will be focussed on developing new classification and visualization tools for our Resistance Gene Identifier (RGI), plus extending the RGI towards k-mer approaches for predicting pathogen-of-origin for metagenomics antimicrobial resistance gene sequences.
Today we say farewell to Arjun Sharma & Suman Virdee. Arjun joined the lab as a second year volunteer, staying to perform a Biochem 3R06 research project. He has been very active in our Comprehensive Antibiotic Resistance Database project, co-developing our CARD*Shark text mining tools for computer-guided curation of literature in PubMed, pipelines for our clinical isolate genome sequencing work, and developing novel algorithms for predicting glycopeptide resistance from genome assemblies. He was the recipient of an IIDR Summer Student Fellowship and leaves the Biochemistry program to enter medical school at the University of Toronto. Suman joined the lab in the 4th year of the Biomedical Discovery & Commercialization program, performing her thesis research on RNA-Seq bioinformatics workflows in a collaboration between our lab and the laboratory of Dr. Kristen Hope (McMaster Stem Cell and Cancer Research Institute), extending her research into the summer by winning a CIHR Summer Undergraduate Research Award. Suman finished her degree and this September starts in the McMaster Master of Science in Global Health program. Bon chance Suman & Arjun!
We haven’t been travelling much this year, but our collaborators have been busy!
Dearborn, D.C., A.B. Gager, A.G. McArthur, M.E. Gilmour, E. Mandzhukova, R.A. Mauck. 2017. How to get diverse MHC genotypes without disassortative mating. Presentation at the 2017 Annual Meeting of the Society for Integrative and Comparative Biology, New Orleans, Louisiana.
McLean, M., D. Theriault, M. Kelley, B.A. Lago, A.G. McArthur, & L. Williams. 2017. Role of Nfe2 and pro-oxidant exposure in inner ear development in zebrafish. Presentation at the Society of Toxicology 56rd Annual Meeting, Baltimore, Maryland.
Williams, L.M., B.A. Lago, A.G. McArthur, A.R. Raphenya, N. Pray, N. Saleem, S. Salas, K. Paulson, R.S. Mangar, Y. Liu, A.H. Vo, & J.A. Shavit. 2017. The transcription factor, Nuclear factor, erythroid 2 (Nfe2), is a regulator of the oxidative stress response during Danio rerio development. Presentation at the Society of Toxicology 56rd Annual Meeting, Baltimore, Maryland.
Winsor, G.L., C. Bertelli, K.K. Tsang, B. Alcock, A.G. McArthur, & F.S.L. Brinkman. 2017. Pseudomonas Genome Database 2017: Improved gene/AMR/VF/genomic island annotations, comparative genome analyses, and a platform for facilitating public health genomic epidemiology. Presentation at the 16th International Conference on Pseudomonas, Liverpool, United Kingdom.
Anastasia joins us & the IIDR for Summer 2017 from the Center for Genomic Sciences, UNAM, Cuernavaca, Mexico as part of her successful competition for a Mitacs Globalink Internship. Throughout the summer, she will be working on data and algorithm development for antimicrobial resistance genomic surveillance. Welcome Anastasia!
Well done Kara! The OGS is a provincial merit-based scholarship, with awards available to graduate students in all disciplines of academic study, while the MacDATA Graduate Fellowship Program is part of McMaster’s new MacDATA Institute. Learn more about Kara’s Masters research on antimicrobial resistance genomics here.
Congratulations Arjun (top right)! See the details here.
Congratulations to our 3rd year Biochemistry & Biomedical Sciences students upon completion of their research projects! Both will be staying on as summer research students.
Tammy Lau – The Importance of Protocol Design for RNA-Seq in the Ongoing Development of a Novel Technology for Tissue Specific Gene Expression Profiling in C. elegans
Arjun Sharma – The Glycopeptide Resistance Predictor