Table of Contents |
guest 2024-04-20 |
Event Information
Speakers
Susan Abbatiello
Michael MacCoss
Matthew MacDonald
Brendan MacLean
Sarah Michaud
Bhavin Patel
Pawel Sadowski
Birgit Schilling
Tobias Schmidt
Selene Swanson
Sebastian Vaca
Thanks!
Thanks for helping us celebrate the 10th anniversary of the first release (in 2009) of Skyline at ASMS!
The celebration at our eighth annual Skyline User Group Meeting was held at the incomparable Georgia Aquarium on June 2, 2019. Enthusiasm ran high among the 250 attendees (a new record!) with standing room only both for lunch and the presentations. A 30' glass viewing window into one of the aquarium's largest saltwater displays with 20' whale sharks circling made even a standing lunch worth the visit. The aquatic theme continued into the presentation space with its aqua-toned walls and oceanic ceiling architecture (no sharks allowed) where nine speakers covered a diverse range of topics from developments in DIA peptide transition selection, analysis methods for metabolomics data collected on GCMS instruments and research of modified ribonucleosides in tRNA and mRNA -- all with Skyline as a critical research tool. Two longtime proteomics researchers who lead the original CPTAC studies that motivated Skyline development from the start -- Sue Abbatiello (Northeastern) and Birgit Schilling (Buck Institute) gave an informative and humorous retrospective of Skyline's early years up to the present. The original and now principal developer of Skyline (now a team of 9!) capped off the afternoon with his thoughts on 10 years of Skyline and his vision for 10 more years to come.
As our 10th anniversary gift, every attendee received a Skyline logoed sweatshirt "hoodie" in thanks.
All sessions were recorded and can be accessed, along with presentation PDFs, under the links for each speaker below.
The Skyline Team would once again like to thank all nine speakers for preparing and giving up their Sunday afternoon before ASMS to present at our forum. We'd also like to thank our sponsors for their continued financial support which allows us to keep our User Group Meeting free to participants. Lastly, we'd like to thank all our users for continuing to use Skyline and constantly pushing us to improve, adapt and make Skyline the best tool for your ongoing scientific research. Your enthusiasm for and support of Skyline has made the last 10 years incredibly rewarding.
Thank you! -- Brendan MacLean and Mike MacCoss Event Organizers
Michael J. MacCoss, Ph.D. (University of Washington): Introduction and event host
Full Talks
Brendan MacLean (MacCoss Lab, University of Washington): Status of the Skyline open-source software project 10 years after its inception
The Skyline project started just after ASMS 2008 as a 2-year effort to bring better SRM/MRM software tools to the NCI-CPTAC Verification Working Group that could support the variety of mass spectrometers in use in participating laboratories. Nearly 10 years later, the Skyline project is a thriving proteomics community open-source collaboration supporting 6 mass spec instrument vendors integrated with a wide variety of external software, with thousands of users worldwide and many thousands of instances started each week. (More info...)
Birgit Schilling, Ph.D. (Buck Insitute) and Susan Abbatiello, Ph.D. (Northeastern): Skyline: 10-year Retrospective: Part 1; Part 2
Ten years ago a group of about 20 dedicated triple quadrupole operators met at the Broad Institute to learn about the features of Skyline -- this modest beginning evolved into the Skyline User Group Meeting -- the reason you are here today. Nowadays, the Skyline user base has grown significantly with 94,000 new installations of Skyline since 2009.
Pawel Sadowski, Ph.D. (Queensland University of Technology): Teaching Old Dog New Tricks: Adaptation of Skyline to Analyze Untargeted Metabolomics Data Collected on GCMS Instrument
Processing untargeted metabolomics data collected on GCMS instruments comes with specific challenges and often requires specialized (or expensive) data analysis software. (More info...)
Tobias Schmidt, (Technical University Munich): Using Prosit for PRM assay development and optimization
We propose a novel cost-efficient approach which utilized our deep learning framework Prosit for the generation of in-silico spectral libraries with near reference data quality for virtually any peptide on a proteome-wide scale. (More info...)
Selene Swanson, (Stowers Institute): <a=href="https:>Absolute quantitative analysis of modified ribonucleosides in tRNA and mRNA using Skyline
Epitranscriptomics is a link between epigenomics and proteomics. Understanding the role of RNA modifications is important in elucidating many fundamental cellular processes such as RNA structural stability, splicing, and translation efficiency. Using a Lumos™ Tribrid™ Mass Spectrometer and Skyline, modified ribonucleosides (RNs) in E. coli tRNA and mammalian mRNA were identified and quantified based on calibration curves generated from known standard RNs. (More info...)
Sebastian Vaca, Ph.D. (Broad Institute): <a=href="https:>Avant-garde: A Skyline External Tool for automated data-driven DIA data curation.
Developments in Data-independent Acquisition data analysis have enabled the detection of large numbers of peptides. However, most tools focus on statistical validation of peptide detection (using target/decoy approaches) but do not address the quantitative suitability of the signals extracted. (More info...)
Event Information
Welcome
The Skyline Team is pleased to announce the Eighth Annual Skyline User Group Meeting, which will be held in Atlanta, GA on Sunday afternoon before ASMS. This year we are also celebrating Skyline's 10th birthday. We plan to make this year's event extra special -- so be sure to register and attend! (Check out the coveted Skyline-branded hoodies as you register.) We would like to thank the event sponsors (see below) for their generosity and interest in collaborating with the Skyline project on exciting new targeted and quantitative mass spec techniques.
--Brendan
When: Sunday, June 2, 2019
12:00 - 1:00 pm : Lunch served
1:00 - 2:30 pm : Presentations
2:30 - 3:00 pm : Snacks and break-out discussions
3:00 - 4:30 pm : Presentations
Where: Georgia Aquarium 246 Ivan Allen Jr. Blvd., Atlanta, GA 30313 (map)
- From the Georgia World Conference Building A, walk outside to Andrew Young International Blvd and head east towards Marietta Street
- Turn left on Marietta St. NW and walk three blocks to Ivan Allen Jr. Blvd
- Make a right turn at the crosswalk to heading towards Luckie Street -- the aquarium will be in front of you on the right side of Ivan Allen Jr. Blvd
- Just past the parking entrance, follow a blue walkway to the Ocean's Entrance
- From the Ocean's Ballrooms entrance, proceed to the Pacific Ballroom
[registration closed]
We have closed down our registration process but you can still attend! Please just come to Georgia Aquarium for "day of" registration.
Confirmed Speakers
Michael J. MacCoss, Ph.D. (University of Washington): Introduction and event host
Brendan MacLean (MacCoss Lab, University of Washington): Status of the Skyline open-source software project 10 years after its inception
The Skyline project started just after ASMS 2008 as a 2-year effort to bring better SRM/MRM software tools to the NCI-CPTAC Verification Working Group that could support the variety of mass spectrometers in use in participating laboratories. Nearly 10 years later, the Skyline project is a thriving proteomics community open-source collaboration supporting 6 mass spec instrument vendors integrated with a wide variety of external software, with thousands of users worldwide and many thousands of instances started each week. (More info...)
Susan Abbatiello, Ph.D. (Northeastern) and Birgit Schilling, Ph.D. (Buck Insitute): Skyline: 10-year Retrospective
Ten years ago a group of about 20 dedicated triple quadrupole operators met at the Broad Institute to learn about the features of Skyline -- this modest beginning evolved into the Skyline User Group Meeting -- the reason you are here today. Nowadays, the Skyline user base has grown significantly with 94,000 new installations of Skyline since 2009. (More info...)
Pawel Sadowski, Ph.D. (Queensland University of Technology): Teaching Old Dog New Tricks: Adaptation of Skyline to Analyze Untargeted Metabolomics Data Collected on GCMS Instrument
Processing untargeted metabolomics data collected on GCMS instruments comes with specific challenges and often requires specialized (or expensive) data analysis software. (More info...)
Tobias Schmidt, (Technical University Munich): Using Prosit for PRM assay development and optimization
We propose a novel cost-efficient approach which utilized our deep learning framework Prosit for the generation of in-silico spectral libraries with near reference data quality for virtually any peptide on a proteome-wide scale. (More info...)
Selene Swanson, (Stowers Institute): <a=href="https:>Absolute quantitative analysis of modified ribonucleosides in tRNA and mRNA using Skyline
Epitranscriptomics is a link between epigenomics and proteomics. Understanding the role of RNA modifications is important in elucidating many fundamental cellular processes such as RNA structural stability, splicing, and translation efficiency. Using a Lumos™ Tribrid™ Mass Spectrometer and Skyline, modified ribonucleosides (RNs) in E. coli tRNA and mammalian mRNA were identified and quantified based on calibration curves generated from known standard RNs. (More info...)
Sebastian Vaca, Ph.D. (Broad Institute): <a=href="https:>Avant-garde: A Skyline External Tool for automated data-driven DIA data curation.
Developments in Data-independent Acquisition data analysis have enabled the detection of large numbers of peptides. However, most tools focus on statistical validation of peptide detection (using target/decoy approaches) but do not address the quantitative suitability of the signals extracted. (More info...)
Speakers
Ten speakers with interesting and different areas of expertise in Skyline use and development spoke at the Skyline User Group Meeting at ASMS 2019 in Atlanta, GA.
* PDFs of the presentations are posted on each speaker's page below:
Susan Abbatiello
Susan Abbatiello Ph.D., received a Ph.D. in analytical chemistry at the University of Florida, and completed a postdoc at the University of Pittsburgh’s Hillman Cancer Center. Susan is currently the executive director of the MS Core Facility at the Barnett Institute at Northeastern University, and a visiting scientist in the Proteomics Platform at the Broad Institute of MIT and Harvard. Susan’s research focuses on the development of experiments to measure potential protein biomarkers in cell lines and blood for diseases such as cancer. Susan has co-chaired a committee that is part of the National Cancer Institute’s Clinical Proteomic Technology Assessment for Cancer (CPTAC).
Skyline: 10-year Retrospective
Ten years ago a group of about 20 dedicated triple quadrupole operators met at the Broad Institute to learn about the features of Skyline -- this modest beginning evolved into the Skyline User Group Meeting -- the reason you are here today. Nowadays, the Skyline user base has grown significantly with 94,000 new installations of Skyline since 2009. Birgit and I will provide an entertaining retrospective over the last 10 years – what were its key achievements? Read More[PDF]
Michael MacCoss
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Michael MacCoss Mike became interested in biomedical applications of mass spectrometry while working in Dr. Patrick Griffin’s protein mass spectrometry lab at Merck Research Laboratories. He obtained a Ph.D. with Professor Dwight Matthews and pursued a postdoc with Professor John R. Yates III. In 2004 he started the MacCoss lab at the University of Washington and it became obvious that while mass spectrometry data could be collected quickly and robustly, the lack of computational tools for the visualization and analysis of these data was a stumbling block. In 2008 he hired Brendan MacLean with the goal of developing professional quality software tools for quantitative proteomics.
Read More
Mike has worked closely with the Skyline development team and our outstanding group of laboratory scientists and collaborators to ensure that our software uses analytical approaches that have been thoroughly vetted by the mass spectrometry community.
[PDF] |
Matthew MacDonald
Matthew MacDonald ,Ph.D. Throughout his graduate and postdoctoral training to his current position as an assistant professor of psychiatry, University of Pittsburgh, MacDonald's main research focus has been validating and utilizing proteomic approaches to investigate synaptic protein network dysfunction in postmortem brain tissue from patients with neuropsychiatric disease. As a postdoctoral fellow (with Drs. Robert Sweet and Nathan Yates) and now a faculty member he has continued to push the envelope of proteomics approaches in human postmortem brain tissue to increase proteomic depth, throughput and spatial resolution.
Multi-omics Approach Identifies Pathological Phosphorylation Events Driving Synapse Loss in Schizophrenia
Schizophrenia is a complex trait disorder in which genetic risk factors converge on synaptic protein networks, altering synaptic architecture, and impairing brain function. Synaptic protein network features include protein expression, trafficking, and activity. To assess interactions between these features, we utilized microscopy as well as targeted and label free mass spectrometry to quantify synapse number and size as well as protein expression, synaptic protein levels, and protein phosphorylation in cortical tissue from 50 schizophrenia and 50 matched control subjects. Read More[PDF]
Brendan MacLean
Brendan MacLean worked at Microsoft for 8 years in the 1990s where he was a lead developer and development manager for the Visual C++/Developer Studio Project. Since leaving Microsoft, Brendan has been the Vice President of Engineering for Westside Corporation, Director of Engineering for BEA Systems, Inc., Sr. Software Engineer at the Fred Hutchinson Cancer Research Center, and a founding partner of LabKey Software. In this last position he was one of the key programmers responsible for the Computational Proteomics Analysis System (CPAS), made significant contributions to the development of X!Tandem and the Trans Proteomic Pipeline, and created the LabKey Enterprise Pipeline. Since that time, he has worked as a Sr. Software Engineer within the MacCoss lab and lead all aspects of design, development and support in creating the Skyline Targeted Proteomics Environment and its growing worldwide user community.
Status of the Skyline open-source software project 11 years after its inception
The Skyline project started just after ASMS 2008 as a 2-year effort to bring better SRM/MRM software tools to the NCI-CPTAC Verification Working Group that could support the variety of mass spectrometers in use in participating laboratories. Nearly 11 years later, the Skyline project is a thriving proteomics community open-source collaboration supporting 6 mass spec instrument vendors, integrated with a wide variety of external software, with thousands of users worldwide and many thousands of instances started each week. Read More- New! Audit logging
- New! Small molecule user interface
- Advances in calibrated quantification
- Improved command-line support running on Linux
- Growth in the Skyline software ecosystem for targeted MS (Skyline, Panorama, and External Tools)
- Strong industry support from instrument vendors
- Courses, workshops, webinars and other efforts to create instructional resources for the Skyline community
[PDF]
Sarah Michaud
Sarah Michaud received her M.Sc. in Genome Sciences and Technology at the University of British Columbia, where she investigated quantitative proteomics of honeybees during viral infection. She began her work developing targeted assays for quantitation of mouse plasma proteins at MRM Proteomics in 2014, and is now a project manager at the UVic Genome BC Proteomics Center in Victoria, Canada, where her focus is the development of 3,000 MRM assays in 20+ mouse tissues.
Development of Quantitative MRM Assays for the Measurement of 3,000 Proteins across 20 Mouse Tissues
An ongoing effort at our Centre is the development of MRM-MS assays quantifying 3,000 proteins across 20 mouse tissues for use in molecular phenotyping. Our assays use stable isotope-labelled standard peptides for quantitation and undergo rigorous optimization of peptide- and instrument-specific parameters followed by assay development and validation according to CPTAC guidelines. Assay development determines the LLOQ and linear range of the assay via standard curve, and assay variability is calculated over 5 days at three concentrations spanning the final range of the assays. Read More[PDF]
Bhavin Patel
Bhavin Patel , MD. works as a senior research and development scientist in MS reagents group at Thermo Fisher Scientific. I have a medical Doctor degree from the MS University in India, and a masters in biotechnology from the Georgetown University. I had led a protein biomarker targeted MS assay group at NextGen Sciences and a proteomics/mass spectrometry group at Fox Chase Cancer Center before joining Thermo Fisher Scientific in 2013. Since joining Thermo, I have led the development of targeted MS assay kits, new MS system suitability standards, protein sample preparation reagents, antibody validation method for IP-MS, and reagents for targeted quantitative proteomic analysis.
Targeted Mass Spectrometry Assays for Absolute Quantitation of AKT/mTOR Signaling Pathway Proteins
Proteomics offers a deeper understanding of the protein events influencing cellular and biological function. However, Identifying key regulated proteins and their implicated signaling pathways represents a major analytical challenge to researchers. We have optimized a multiplex immunoprecipitation to targeted mass spectrometry (mIP-tMS) workflow to develop the SureQuant pathway panels, achieving simultaneous enrichment and absolute quantitation of multiple total and phosphorylated proteins from the AKT/mTOR pathway. Read More[PDF]
Pawel Sadowski
Pawel Sadowski, Ph.D. completed his Ph.D. in the field of quantitative proteomics at the University of Cambridge. During subsequent post-doctoral appointments at the New York University and at the University of New South Wales in Sydney he applied label-free and label-based approaches to study biomarkers, protein-protein interactions and PTMs. In 2013 Pawel joined Queensland University of Technology in Brisbane to lead Proteomics and Small Molecule Mass Spectrometry Core in the QUT Central Analytical Research Facility. He supervises research that utilizes LCMS and GCMS instrumentation. Pawel’s interest lays in leveraging the power of recent advancements in data acquisition strategies for MS-based proteomics to drive clinical veterinary research into the next generation. More recently, he has started using quantitative mass spectrometry to study metabolites and other small molecules.
Teaching Old Dog New Tricks: Adaptation of Skyline to Analyze Untargeted Metabolomics Data Collected on GCMS Instrument
Processing untargeted metabolomics data collected on GCMS instruments comes with specific challenges and often requires specialized (or expensive) data analysis software. Moreover, metabolomics researchers tend to apply laboratory-specific GCMS data processing criteria depending on the functionalities available in their favorite software package which prevents others from being able to reproduce the results if they do not have access to the same tools. Read More[PDF]
Birgit Schilling
Birgit Schilling, Ph.D. is assistant professor and the director of the Mass Spectrometry Core at the Buck Institute for Research on Aging. She is particularly interested in protein chemistry and modern mass spectrometric technologies to advance the field of proteomics mainly in the context of aging research. Her several research projects include investigation of protein phosphorylation, glycosylation, acetylation, and other posttranslational modifications. We routinely perform quantitative proteomics studies investigating differential expression of intracellular proteins as well as analyzing protein secretomes during disease and aging processes. My group has adopted several novel proteomic technologies and are using proteomic data-independent acquisitions (DIA), or SWATH which allows us to accurately determine changes in relative protein expression level between multiple different conditions. In addition, she has intensively used label-free protein quantification approaches (MS1 Filtering), as well as newer quantitative and highly comprehensive workflows, such as high-resolution data-independent acquisitions (DIA or SWATH Quantification), and parallel reaction monitoring (PRM or high resolution MRM).
Skyline: 10-year Retrospective
Ten years ago a group of about 20 dedicated triple quadrupole operators met at the Broad Institute to learn about the features of Skyline -- this modest beginning evolved into the Skyline User Group Meeting -- the reason you are here today. Nowadays, the Skyline user base has grown significantly with 94,000 new installations of Skyline since 2009. Sue and I will provide an entertaining retrospective over the last 10 years – what were its key achievements? Read More[PDF]
Tobias Schmidt
Tobias Schmidt is a fourth-year Ph.D. candidate at the chair of Proteomics and Bioanalytics of Prof. Kuster in Freising, Technical University Munich, Germany. Before coming to the Technical University of Munich for his MSc in molecular biotechnology he spent three years at the Karlsruhe Institute of Technology studying mathematics.
His research interest is in in-memory databases (ProteomicsDB), combining modern machine learning with high quality synthetic and real-world data as well as porting legacy (academic) systems to new technologies. His doctoral thesis work explores among other things the usage of his prediction tools for data analysis pipelines requiring high-quality spectral libraries.
Using Prosit for PRM assay development and optimization
Setting up and analyzing MRM/PRM assays require prior information on the retention times and fragmentation spectra of peptides. For so far unobserved peptides, synthetic peptides can be used to obtain these characteristics, however, are costly with uncertain results. We propose a novel cost-efficient approach which utilized our deep learning framework Prosit for the generation of in-silico spectral libraries with near reference data quality for virtually any peptide on a proteome-wide scale. Read More[PDF]
Selene Swanson
Selene Swanson's professional career started in the late 80’s at the Microchemical Facility, University of Minnesota as a junior scientist primarily responsible for protein sequencing, peptide synthesis, DNA sequencing/synthesis and amino acid analysis. After a journey through oncology, hematology, gene therapy and nephrology, she landed in the field of proteomics in the early 2000’s at the Proteomics Center of the Stowers Institute for Medical Research. In 2016, after learning the ropes in MudPIT, she was asked to explore the small molecules arena, specifically modified ribonucleosides, given the availability of state-of-the-art mass spectrometers in the center. With the limited open resources available for analyzing PRM data from small molecules, Skyline has proven to be an invaluable resource to meet our needs.
Absolute quantitative analysis of modified ribonucleosides in tRNA and mRNA using Skyline
Epitranscriptomics is a link between epigenomics and proteomics. Understanding the role of RNA modifications is important in elucidating many fundamental cellular processes such as RNA structural stability, splicing, and translation efficiency. Using a Lumos™ Tribrid™ Mass Spectrometer and Skyline, modified ribonucleosides (RNs) in E. coli tRNA and mammalian mRNA were identified and quantified based on calibration curves generated from known standard RNs. The detection levels of the four 2-O’-methylated RNs ranged from undetectable to 0.7ng and 1.8ng out of 500ng each for mammalian mRNA and tRNA, respectively. Read More[PDF]
Sebastian Vaca
Sebastian Vaca , Ph.D. received his Master’s degree in Analytical Sciences from the University of Strasbourg, France. Afterward, he joined the BioOrganic Mass Spectrometry Laboratory (LSMBO, Strasbourg) to work with Dr. Alain Van Dorsselaer and Dr. Christine Carapito. His research aimed at improving the proteome characterization by quantitative mass spectrometry and proteogenomics method development. He received his Ph.D. in June 2016 and joined Dr. Jacob Jaffe’s lab at The Broad Institute, Cambridge, MA. He focuses on Data-Independent Acquisition method development and data analysis in order to understand phosphosignaling in cancerous cells. Sebastian’s motivation to develop an automated data-curation tool came from the hundreds of hours of sitting in front of a computer manually integrating chromatographic peaks.
Avant-garde: A Skyline External Tool for automated data-driven DIA data curation
Developments in Data-independent Acquisition data analysis have enabled the detection of large numbers of peptides. However, most tools focus on statistical validation of peptide detection (using target/decoy approaches) but do not address the quantitative suitability of the signals extracted. In practice, time-consuming manual signal curation is still required for rigorous quantitation. Here we introduce Avant-Garde (AvG), a Skyline External Tool for automated MS data curation meant to polish the results of DIA/PRM analysis tools and assess the quantitative suitability of DIA signals. Read More[PDF]