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Fondrie, W. E.

Publications and source records attributed to Fondrie, W. E..

15 recordsLinked to original sources

Structure-free, site-resolved contrastive learningextends small-molecule discovery beyond the reachof structure-based modeling

Virtual screening asks which molecules, among an enormous space of drug-like chemistry, are worth synthesizing and testing against a protein target. Most modern methods answer by building and scoring an explicit three-dimensional pose through molecular docking, or the co-folding models that now approach experimental accuracy. Building these poses presumes a well-defined pocket, and the non-orthosteric, cryptic, and intrinsically disordered sites where unexplored ligandability lies offer none. There, these methods fail to generalize. Here we present Ptarmigan-1, a contrastive model that co-embeds the residues of a protein with candidate small molecules in a shared latent space, from sequence and two-dimensional chemistry alone, and without ever constructing a pose. Engagement reduces to the proximity of precomputed embeddings. Freed from the pose, Ptarmigan-1 trains directly on chemoproteomic and bioactivity data of mixed resolution, scores a compound in ten milliseconds rather than the tens of seconds a co-folding model demands, and resolves each prediction to the residues a compound engages. On well-folded, orthosteric targets it performs comparably to a collection of co-folding and docking models, and on covalent, cryptic, and disordered sites it matches or exceeds them. It localizes reversible and covalent inhibitors to the pockets they engage, even for targets withheld from training, and screens the entire human proteome against a library of 3.4 billion compounds in under a day. By decoupling molecular recognition from structure, Ptarmigan-1 recasts virtual screening as a reusable index that continuously improves as data accumulate.

molecular biology↗

Improvements to Casanovo, a deep learning de novo peptidesequencer

Casanovo is a state-of-the-art deep learning model for de novo peptide sequencing from mass spectrometry proteomics data. Here we report on a series of enhancements to Casanovo, aimed at improving the interpretability of the scores assigned to predicted peptides, generalizing the software for use in database search, speeding up training and prediction runtimes, and providing workflows and visualization tools to facilitate adoption of Casanovo and interpretation of its results. Our goal is to make Casanovo accurate and easy to use for applications such as metaproteomics, antibody sequencing, immunopeptidomics, and discovery of novel peptide sequences in standard proteomics analyses. Casanovo is available as open source at https://github.com/Noble-Lab/casanovo.

molecular biology↗

Native, Spatiotemporal Profiling of the Global Human Regulome

The regulome, comprising transcription factors, cofactors, chromatin remodelers, and other regulatory proteins, forms the core machinery by which cells interpret signals and execute gene expression programs. Despite its central role in development, disease, and drug response, the regulome remains largely uncharted at scale due to its dynamic, low-abundance, and chromatin-associated nature. Here, we present a method for scalable, regulome profiling for global, compartment-resolved quantification of native regulome proteins. By enriching DNA- and chromatin-associated proteins and profiling them using high-throughput, label-free DIA mass spectrometry, regulome profiling captures chromatin-associated proteins across 36 human cell lines and thousands of perturbations. The resulting Regulome Atlas recovers nearly 60% of known human transcription factors, reveals lineage-specific TF localization, and distinguishes active nuclear engagement from latent, unbound states. We demonstrate that regulome profiles resolve acute immune pathway activation prior to transcriptional changes, identify previously unrecognized drug-induced regulome responses, and enable proteome-scale readouts of compound target engagement and complex remodeling. This work establishes a foundational resource for decoding the regulatory proteome and provides a blueprint for integrating regulome data into next-generation models of cellular behavior.

cell biology↗

A haplotype-resolved view of human gene regulation

Diploid human cells contain two non-identical genomes, and differences in their regulation underlie human development and disease. We present Fiber-seq Inferred Regulatory Elements (FIRE) and show that FIRE provides a more comprehensive and quantitative snapshot of the accessible chromatin landscape across the 6 Gbp diploid human genome, overcoming previously unrecognized biases in existing regulatory element catalogs. FIRE enables comprehensive detection of haplotype-selective chromatin accessibility (HSCA), exposing novel imprinted elements lacking underlying parent-of-origin CpG methylation differences, and gene regulatory modules that permit genes to escape X chromosome inactivation. We uncover that the human leukocyte antigen (HLA) locus harbors the most HSCA in immune cells, where we resolve specific transcription factor (TF) binding events disrupted by disease-associated variants. Finally, we demonstrate that the regulatory landscape of a cell is littered with autosomal somatic chromatin epimutations that are propagated by clonal expansions to create mitotically stable and non-genetically deterministic chromatin alterations.

genomics↗

Target-decoy false discovery rate estimation using Crema

Assigning statistical confidence estimates to discoveries produced by a tandem mass spectrometry proteomics experiment is critical to enabling principled interpretation of the results and to assess the cost/benefit ratio of experimental follow-up. The most common technique for computing such estimates is to use target-decoy competition (TDC), in which observed spectra are searched against a database of real (target) peptides and a database of shuffled or reversed (decoy) peptides. TDC procedures for estimating the false discovery rate (FDR) at a given score threshold have been developed for application at the level of spectra, peptides, or proteins. Although these techniques are relatively straightforward to implement, it is common in the literature to skip over the implementation details or even to make mistakes in how the TDC procedures are applied in practice. Here we present Crema, an open source Python tool that implements several TDC methods of spectrum-, peptide- and protein-level FDR estimation. Crema is compatible with a variety of existing database search tools and provides a straightforward way to obtain robust FDR estimates.

bioinformatics↗

Evaluating proteomics imputation methods with improved criteria

Quantitative measurements produced by tandem mass spectrometry proteomics experiments typically contain a large proportion of missing values. This missingness hinders reproducibility, reduces statistical power, and makes it difficult to compare across samples or experiments. Although many methods exist for imputing missing values in proteomics data, in practice, the most commonly used methods are among the worst performing. Furthermore, previous benchmarking studies have focused on relatively simple measurements of error, such as the mean-squared error between the imputed and the held-out observed values. Here we evaluate the performance of a set of commonly used imputation methods using three practical, "downstream-centric" criteria, which measure the ability of imputation methods to reconstruct differentially expressed peptides, identify new quantitative peptides, and improve peptide lower limit of quantification. Our evaluation spans several experiment types and acquisition strategies, including datadependent and data-independent acquisition. We find that imputation does not necessarily improve the ability to identify differentially expressed peptides, but that it can identify new quantitative peptides and improve peptide lower limit of quantification. We find that MissForest is generally the best performing method per our downstream-centric criteria. We also argue that exisiting imputation methods do not properly account for the variance of peptide quantifications and highlight the need for methods that do.

bioinformatics↗

Sequence-to-sequence translation from mass spectra to peptides with a transformer model

A fundamental challenge for any mass spectrometry-based proteomics experiment is the identification of the peptide that generated each acquired tandem mass spectrum. Although approaches that leverage known peptide sequence databases are widely used and effective for well-characterized model organisms, such methods cannot detect unexpected peptides and can be impractical or impossible to apply in some settings. Thus, the ability to assign peptide sequences to the acquired tandem mass spectra without prior information--de novo peptide sequencing--is valuable for gaining biological insights for tasks including antibody sequencing, immunopeptidomics, and metaproteomics. Although many methods have been developed to address this de novo sequencing problem, it remains an outstanding challenge, in part due to the difficulty of modeling the irregular data structure of tandem mass spectra. Here, we describe Casanovo, a machine learning model that uses a transformer neural network architecture to translate the sequence of peaks in a tandem mass spectrum into the sequence of amino acids that comprise the generating peptide. We train a Casanovo model from 30 million labeled spectra and demonstrate that the model outperforms several state-of-the-art methods on a cross-species benchmark dataset. We also develop a version of Casanovo that is fine-tuned for non-enzymatic peptides. Finally, we demonstrate that Casanovos superior performance improves the analysis of immunopeptidomics and metaproteomics experiments and allows us to delve deeper into the dark proteome.

bioinformatics↗

The Crux toolkit for analysis of bottom-up tandem mass spectrometry proteomics data

The Crux tandem mass spectrometry data analysis toolkit provides a collection of algorithms for analyzing bottom-up proteomics tandem mass spectrometry data. Many publications have described various individual components of Crux, but a comprehensive summary has not been published since 2014. The goal of this work is to summarize the functionality of Crux, focusing on developments since 2014. We begin with empirical results demonstrating our recently implemented speedups to the Tide search engine. Other new features include a new score function in Tide, two new confidence estimation procedures, as well as three new tools: Param-medic for estimating search parameters directly from mass spectrometry data, Kojak for searching cross-linked mass spectra, and DIAmeter for searching data independent acquisition data against a sequence database.

bioinformatics↗

nf-encyclopedia: A cloud-ready pipeline for chromatogram library data-independent acquisition proteomics workflows

Data independent acquisition (DIA) mass spectrometry methods provide systematic and comprehensive quantification of the proteome; yet, relatively few open-source tools are available to analyze DIA proteomics experiments. Fewer still are tools that can leverage gas phase fractionated (GPF) chromatogram libraries to enhance the detection and quantification of peptides in these experiments. Here, we present nf-encyclopedia, an open-source NextFlow pipeline that connects three open-source tools--MSConvert, EncyclopeDIA, and MSstats--to analyze DIA proteomics experiments with or without chromatogram libraries. We demonstrate that nf-encyclopedia is reproducible both when run on a cloud platform or a local workstation and provides robust peptide and protein quantification. Additionally, we found that MSstats enhances protein-level quantitative performance over EncyclopeDIA alone. Finally, we benchmarked the ability nf-encyclopedia to scale to large experiments in the cloud by leveraging the parallelization of compute resources. The nf-encyclopedia pipeline is available under a permissive Apache 2.0 license--run it on your desktop, cluster, or in the cloud: https://github.com/TalusBio/nf-encyclopedia.

bioinformatics↗

Semi-supervised machine learning for sensitive open modification spectral library searching

A key analysis task in mass spectrometry proteomics is matching the acquired tandem mass spectra to their originating peptides by sequence database searching or spectral library searching. Machine learning is an increasingly popular post-processing approach to maximize the number of confident spectrum identifications that can be obtained at a given false discovery rate threshold. Here, we have integrated semi-supervised machine learning in the ANN-SoLo tool, an efficient spectral library search engine that is optimized for open modification searching to identify peptides with any type of post-translational modification. We show that machine learning rescoring boosts the number of spectra that can be identified for both standard searching and open searching, and we provide insights into relevant spectrum characteristics harnessed by the machine learning model. The semi-supervised machine learning functionality has now been fully integrated into ANN-SoLo, which is available as open source under the permissive Apache 2.0 license on GitHub at https://github.com/bittremieux/ANN-SoLo.

bioinformatics↗

De novo mass spectrometry peptide sequencing with a transformer model

Tandem mass spectrometry is the only high-throughput method for analyzing the protein content of complex biological samples and is thus the primary technology driving the growth of the field of proteomics. A key outstanding challenge in this field involves identifying the sequence of amino acids--the peptide--responsible for generating each observed spectrum, without making use of prior knowledge in the form of a peptide sequence database. Although various machine learning methods have been developed to address this de novo sequencing problem, challenges that arise when modeling tandem mass spectra have led to complex models that combine multiple neural networks and post-processing steps. We propose a simple yet powerful method for de novo peptide sequencing, Casanovo, that uses a transformer framework to map directly from a sequence of observed peaks (a mass spectrum) to a sequence of amino acids (a peptide). Our experiments show that Casanovo achieves state-of-the-art performance on a benchmark dataset using a standard cross-species evaluation framework which involves testing with spectra with never-before-seen peptide labels. Casanovo not only achieves superior performance but does so at a fraction of the model complexity and inference time required by other methods.

bioinformatics↗

A flexible workflow for building spectral libraries from narrow window data independent acquisition mass spectrometry data

Advances in library-based methods for peptide detection from data independent acquisition (DIA) mass spectrometry have made it possible to detect and quantify tens of thousands of peptides in a single mass spectrometry run. However, many of these methods rely on a comprehensive, high quality spectral library containing information about the expected retention time and fragmentation patterns of peptides in the sample. Empirical spectral libraries are often generated through data-dependent acquisition and may suffer from biases as a result. Spectral libraries can be generated in silico but these models are not trained to handle all possible post-translational modifications. Here, we propose a false discovery rate controlled spectrum-centric search workflow to generate spectral libraries directly from gas-phase fractionated DIA tandem mass spectrometry data. We demonstrate that this strategy is able to detect phosphorylated peptides and can be used to generate a spectral library for accurate peptide detection and quantitation in wide window DIA data. We compare the results of this search workflow to other library-free approaches and demonstrate that our search is competitive in terms of accuracy and sensitivity. These results demonstrate that the proposed workflow has the capacity to generate spectral libraries while avoiding the limitations of other methods.

bioinformatics↗

ppx: Programmatic access to proteomics data repositories

The volume of proteomics and mass spectrometry data available in public repositories continues to grow at a rapid pace as more researchers embrace open science practices. Open access to the data behind scientific discoveries has become critical to validate published findings and develop new computational tools. Here, we present ppx, a Python package that provides easy, programmatic access to the data stored in ProteomeXchange repositories, such as PRIDE and MassIVE. The ppx package can either be used as a command line tool or a Python package to retrieve the files and metadata associated with a project when provided its identifier. To demonstrate how ppx enhances reproducible research, we used ppx within a Snakemake workflow to reanalyze a published dataset with the open modification search tool ANN-SoLo and compared our reanalysis to the original results. We show that ppx readily integrates into workflows and our reanalysis produced results consistent with the original analysis. We envision that ppx will be a valuable tool for creating reproducible analyses, providing tool developers easy access to data for development, testing, and benchmarking, and enabling the use of mass spectrometry data in data-intensive analyses. The ppx package is freely available and open source under the MIT license at: https://github.com/wfondrie/ppx

bioinformatics↗

mokapot: Fast and flexible semi-supervised learning for peptide detection

Proteomics studies rely on the accurate assignment of peptides to the acquired tandem mass spectra--a task where machine learning algorithms have proven invaluable. We describe mokapot, which provides a flexible semi-supervised learning algorithm that allows for highly customized analyses. We demonstrate some of the unique features of mokapot by improving the detection of RNA-cross-linked peptides from an analysis of RNA-binding proteins and increasing the consistency of peptide detection in a single-cell proteomics study.

bioinformatics↗

A machine learning strategy that leverages large datasets to boost statistical power in small-scale experiments

Machine learning methods have proven invaluable for increasing the sensitivity of peptide detection in proteomics experiments. Most modern tools, such as Percolator and PeptideProphet, use semi-supervised algorithms to learn models directly from the datasets that they analyze. Although these methods are effective for many proteomics experiments, we suspected that they may be suboptimal for experiments of smaller scale. In this work, we found that the power and consistency of Percolator results was reduced as the size of the experiment was decreased. As an alternative, we propose a different operating mode for Percolator: learn a model with Per-colator from a large dataset and use the learned model to evaluate the small-scale experiment. We call this a "static modeling" approach, in contrast to Percolators usual "dynamic model" that is trained anew for each dataset. We applied this static modeling approach to two settings: small, gel-based experiments and single-cell proteomics. In both cases, static models increased the yield of detected peptides and eliminated the model-induced variability of the standard dynamic approach. These results suggest that static models are a powerful tool for bringing the full benefits of Percolator and other semi-supervised algorithms to small-scale experiments. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=106 SRC="FIGDIR/small/849331v1_ufig1.gif" ALT="Figure 1"> View larger version (20K): org.highwire.dtl.DTLVardef@9be517org.highwire.dtl.DTLVardef@dd6e3eorg.highwire.dtl.DTLVardef@170e35corg.highwire.dtl.DTLVardef@5ec13e_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioinformatics↗