FROG analysis - a community standard to foster reproducibility and curation of constraint-based models

 

Summary

Constraint-based models are used to investigate metabolism in diverse conditions; particularly genome-scale metabolic models (GEMs) with thousands of reactions provide opportunities to analyze organism-specific metabolism. Community standards for consistent model reconstruction, curation, and sharing are crucial to ensure reproducibility, reliability, and FAIR sharing of the models. MEMOTE, a community tool was developed for standardized quality assessment of the models. Currently, it is not possible to assess whether constraint-based models including Genome-Scale metabolic models (GEM) are reproducible because these models often have multiple solutions and the numerical values are not always enumerated in the published manuscripts. Here we initiate a community effort for standardized assessment of model reproducibility, which is currently lacking.  Reproducibility of results is the cornerstone of science and its assessment is an essential part of the curation of a model in the repository such as BioModels. Following discussions at dedicated breakout sessions at HARMONY2020 (minutes of the meeting), COMBINE2020 (minutes of the meeting), and HARMONY2021 (minutes of the meeting) we have now developed FROG analysis, an ensemble of analysis of constraint-based models to generate standardized numerically reproducible reference dataset (aka FROG report). We have also developed a collection of tools that generates FROG reports in a standardized schema. We propose that the modelers share FROG reports along with their model which can be used by the modeler or curator to independently assess the reproducibility of a model. FROG analysis is currently used in BioModels’ workflow for the curation of constraint-based models along with the MEMOTE test suite. To allow retrospective curation of previously published models, we propose that the model authors submit a miniFROG report in addition to autogenerated FROG report. The miniFROG report is a manually created data table in a standardized schema that lists at least a couple of results described in the manuscript and compares them against the model results from the FROG report. Sharing FROG and miniFROG reports along with model (SBML-FBC) files will enable assessment of reproducibility and curation of models and thereby greatly enhance reuse, extension, integration of constraint-based models for new knowledge generation. 

FROG analysis

A curated model should be able to faithfully reproduce the analysis results. Flux values are commonly reported as results in manuscripts with flux balance analysis of constraint-based models. These values cannot be used to test the reproducibility as often multiple solutions exist for the same. Hence, numerically reproducible results of the analysis are required to verify the reproducibility of the models. Following the discussion in the HARMONY2020 and COMBINE2020 meetings, we recognized the following list of outputs/results of the FBA analysis that are numerical reproducible and can be used for curation.

1) Objective Function Values

The objective function value for a defined set of bounds should be comparable/reproducible.

2) Flux Variability Analysis (FVA)

  • FVA span: min/max of flux should be comparable (for a particular objective function value)
  • These values will have only small numerical differences among software, depending on the set boundary conditions.

3) Gene Deletion Fluxes

  • The systematic deletion of all genes one at a time should provide comparable reference results.

4) Reaction Deletion (extended coverage of reaction network)

  • The systematic deletion of all reactions one at a time should provide comparable reference results.

The above FROG references will be central to assessing the reproducibility of the model and the curation efforts.

FROG test suite

The following tools can be used to generate a FROG report. 

  1. fbc_curation (Developer: Matthias König)

a python package for FROG analysis. Currently it includes two separate implementations of the reference files generation by: 

  • cobrapy based on COBRApy (Constraint-Based Reconstruction and Analysis in Python) available from https://github.com/opencobra/cobrapy/

  • cameo based on Cameo (Cameo—Computer Aided Metabolic Engineering and Optimization) available from https://github.com/biosustain/cameo

Command-line Tool: https://github.com/matthiaskoenig/fbc_curation

Documentation: https://fbc-curation.readthedocs.io/en/latest/index.html

Web implementation: http://runfrog.de/

  1. CBMPy model curator (Developer: Brett Oliver)

Script and web-based implementation of the FROG analysis using CBMPy.

Web implementation: CBMPyWEB 

Command-line Tool:  CBMPy FBC curator https://github.com/matthiaskoenig/fbc_curation

Documentation: link to presentation

  1. fbc_curation_matlab (Developer: Karthik Raman)
    MATLAB/COBRA helper for FROG analysis of FBC models.

    Command-line Tool: https://github.com/RamanLab/fbc_curation_matlab

  2. FLUXER
    a web-based implementation supporting FROG analysis
    Web page: 
    https://fluxer.umbc.edu/

Curation of FBC models in BioModels

BioModels is one of the largest repositories of manually curated models. Model curation involves ensuring the model is (1) encoded in a syntactically valid standard format such as SBML, (2) is and reproducible, and (3) semantically enriched with controlled vocabularies such GO, ChEBI, etc. A model author is expected to submit an SBML model (as main file), FROG report (as additional file), and miniFROG (as additional file) to BioModels. Reproducibility of the constraint-based SBML models submitted to BioModels will be verified using the FROG test suite. Curators at BioModels will independently try to reproduce the FROG report using a tool different from the one used by the modeler. If the results are reproducible, the model will be added to the curated branch of BioModels. The quality of the model will be tested using MEMOTE test suite and the report will be uploaded as an additional file. Furthermore, model-level semantic annotations will be added to the model following MIRIAM guidelines. Refer to Figure 1 for the complete workflow. 

 

Figure 1: Workflow for the curation of constraint-based models in BioModels using FROG 

 

 

minFROG description

While the FROG analysis is helpful for new models, the task of retrospective curation of already published models demands comparison of model performance with results reported in the manuscript. This comparison will ensure that the right model version is shared and it reproduces published results. To facilitate this, we have a "miniFROG" report, which provides a template to enter key observations from the manuscript and the agreement with FROG, which is a check on the reproducibility of the model/correctness of the model version, with respect to the published model.

Following are the fields in the miniFROG:

  • Publication Information

  • Organism Name

  • BioModel Identifier

  • Simulation constraints defined in publication

  • Gene/ reaction involved in the constraint

  • Type of constraints in the simulation

  • Tools used for simulation

  • Results from publication

  • Results predicted from FROG

  • Type of F/R/O/G analysis

  • Line in FROG Report

  • Validation of the simulation (Yes/No)

  • Remarks

miniFROG report should be manually prepared using this template and submitted as an additional file during model submission to BioModels for all previously published models. miniFROG report is optional, but highly recommended and can be submitted as an additional file for unpublished models.

Raman et al. 2005 model (BIOMD0000001046)  is curated using FROG test suite and miniFROG standards. It provides an example for miniFROG report.  Click here to view other examples on miniFROG reports. 

Community manuscript

We are preparing a manuscript “FROG analysis - a community standard to foster reproducibility and curation of constraint-based models” 

If you like to contribute to this community effort and the manuscript, 

  1. Please submit one of your constraint-based models (to BioModels*) along with a FROG report generated using any one of the above FROG tools or your own tool in the standard schema.

Contact: biomodels-cura@ebi.ac.uk for any support on submission. While contacting, use the keyword 'FROG' in the subject line.

* If you have already submitted your model to BioModels, kindly upload the  FROG report as an additional file and request to publish the model again. 

Please fill the following form below
https://docs.google.com/forms/d/e/1FAIpQLSca8t77y_bn85zEbVvKBtylN_UnHSurKBCnPTtMTpyVBnpBZA/viewform

All models submitted to BioModels will be curated using the FROG tools, added to the appropriate branch and highlighted in the manuscript.

  1. Develop a package or web application for FROG analysis.
    Please contact: sheriff@ebi.ac.uk