zenodoopen

Systems-level design principles of metabolic rewiring in an animal

Description: This repository contains all source codes, necessary input and output data, and raw figures and tables for reproducing all figures and results published in the following study:  Xuhang Li#, Hefei Zhang#, Thomas Hodder, Wen Wang, L. Safak Yilmaz, Chad L. Myers, Albertha J.M. Walhout. Systems-level Worm Perturb-Seq reveals design principles of metabolic rewiring. (2025) Nature, in press (# equal contribution) Files: This repository contains the entire project working directory for this publication. To deposit into Zenodo, we have individually zipped each subfolder of the root directory. To reproduce our study, it is advised that one should download the entire Zenodo repository and unzip every zip file to produce the corresponding subfolders. The "root.zip" should be directly unzipped into the root directory of this project (not your system root!!).  The recovered working directory should be like this:  <ul> <li>10_revision/</li> <li>1_QC_dataCleaning/</li> <li>2_DE/</li> <li>3_imageAnalysis/</li> <li>4_networkAnalysis/</li> <li>5_GSA/</li> <li>7_FBA_modeling/</li> <li>data_and_tables_to_publish/</li> <li>input_data/</li> <li>generate_all_tables_for_publication.R</li> <li>prepareTbls_met12.R</li> <li>prepareTbls_metVali1_OCC.R</li> <li>prepareTbls_metVali2_b12.R</li> <li>prepareTbls_metVali3_detox.R</li> <li>prepareTbls.R</li> <li>prepareTbls_SPECIAL.R</li> <li>REWIRING_FIGURE_LOOKUP.xlsx</li> </ul> Please be advised that this repository contains raw codes and data that are not directly related to a figure in our paper. However, they may be useful to generate input used in the analysis of a figure, or to reproduce tables in our manuscript. It may also contain unpublished analyses and figures, which we did not intentionally delete and kept for records.  Usage:  Please refer to the table in below to locate a specific file for reproducing a figure of interest (also availabe in the REWIRING_FIGURE_LOOKUP.xlsx under the root directory).  <table> <tbody> <tr> <td>Figure</td> <td>File</td> <td>Linesa</td> <td>Notes</td> </tr> <tr> <td>Fig. 1b</td> <td>1_QC_dataCleaning/5_supplementary_figure_rewiring.R</td> <td>1-190</td> <td> </td> </tr> <tr> <td>Fig. 1c</td> <td>2_DE/3_1_DE_network_analysis.R</td> <td>1-83</td> <td> </td> </tr> <tr> <td>Fig. 1d</td> <td>11_additional analysis/special RNAi.R</td> <td>entire file</td> <td> </td> </tr> <tr> <td>Fig. 1e</td> <td>2_DE/output/metabolic_GRN_layout2.cys</td> <td>open in Cytoscape</td> </tr> <tr> <td>Fig. 1f</td> <td>2_DE/3_1_DE_network_analysis.R</td> <td>1-71</td> <td> </td> </tr> <tr> <td>Fig. 1g</td> <td>2_DE/3_1_DE_network_analysis.R</td> <td>1-145</td> <td> </td> </tr> <tr> <td>Fig. 2a</td> <td>2_DE/5_2_coexpression_analysis_direct_tree.R</td> <td>1-235</td> <td> </td> </tr> <tr> <td>Fig. 2b</td> <td>2_DE/5_1_1_DE_similarity_analysis.R</td> <td>232-287</td> <td> </td> </tr> <tr> <td>Fig. 2c</td> <td>2_DE/5_1_1_DE_similarity_analysis.R</td> <td>468-765</td> <td>For producing the embedding, see "2_DE/clustering_python/cluster_RNAi_UMAP_DBSCAN.ipynb"</td> </tr> <tr> <td>Fig. 2d</td> <td>2_DE/5_2_coexpression_analysis_direct_tree.R</td> <td>136-181</td> <td> </td> </tr> <tr> <td>Fig. 2e</td> <td>2_DE/5_2_coexpression_analysis_direct_tree.R</td> <td>136-338</td> <td> </td> </tr> <tr> <td>Fig. 2f</td> <td>2_DE/6_2_rewiring_summary_map_PCCtree.R</td> <td>entire file</td> <td> </td> </tr> <tr> <td>Fig. 3b</td> <td>2_DE/4_2_pathway_level_analysis.R</td> <td>2247-2539</td> <td> </td> </tr> <tr> <td>Fig. 3c</td> <td>2_DE/4_2_pathway_level_analysis.R</td> <td>2247-2506</td> <td>the published figure uses level 4 and up direction (2_DE/figures/1_pathway_level_analysis/rxn_and_pathway_rewiring_LEVEL4_up.pdf)</td> </tr> <tr> <td>Fig. 3d</td> <td>2_DE/4_2_pathway_level_analysis.R</td> <td>2183-2235</td> <td> </td> </tr> <tr> <td>Fig. 3e</td> <td>2_DE/4_2_pathway_level_analysis.R</td> <td>1751-2159</td> <td> </td> </tr> <tr> <td>Fig. 3g</td> <td>4_networkAnalysis/2_visualize_example_rewirings.R</td> <td>803-807</td> <td>please load the dependency starting from line 1</td> </tr> <tr> <td>Fig. 3h</td> <td>11_additional analysis/glycine tracing/glycine RNAi tracing.R</td> <td>entire file</td> <td>source data also provided along the paper</td> </tr> <tr> <td>Fig. 3j</td> <td>4_networkAnalysis/2_visualize_example_rewirings.R</td> <td>1-601</td> <td> </td> </tr> <tr> <td>Fig. 3k</td> <td>4_networkAnalysis/1_rewiring_network_modules.R</td> <td>entire file</td> <td> </td> </tr> <tr> <td>Fig. 4a</td> <td>7_FBA_modeling/CR_model_final_run/2_DEG_modeling_basic_CR_model.R</td> <td>853-881</td> <td>run lines 1-100 to load dependency; Euler plot depends on a random seeds; we manually picked one output figure that has best visual representations; see the corresponding Github or "7_FBA_modeling/CR_model_final_run/a1_gene_obj_classification.m" for generating the input matrix (core function matrix)</td> </tr> <tr> <td>Fig. 4b</td> <td>7_FBA_modeling/CR_model_final_run/2_DEG_modeling_supp_gspd_1_example_objHeatmap.R</td> <td>entire file</td> <td> </td> </tr> <tr> <td>Fig. 4c</td> <td>7_FBA_modeling/CR_model_final_run/2_DEG_modeling_basic_CR_model.R</td> <td>1-270</td> <td> </td> </tr> <tr> <td>Fig. 4d</td> <td>7_FBA_modeling/CR_model_final_run/3_DEG_modeling_edge_direction_test_randFBA.R, 7_FBA_modeling/CR_model_final_run/3_DEG_modeling_edge_direction_test_randGRN.R</td> <td>entire file</td> <td>Reproducing this figure is complex because it integrates multiple randomization results; search "fitted_CR_interaction_diagram.pdf" or "fitted_CR_interaction_diagram_randGRN.pdf" to locate the code for making the figure (two randomizations, respectively), and track up for a few lines to load the dependency. To fully reproduce the analysis, follow the instructions in the Github and you will need to execute all four files named "3_DEG_modeling_edge_*.R".</td> </tr> <tr> <td>Fig. 4f</td> <td>7_FBA_modeling/CR_model_final_run/2_DEG_modeling_supp_fused_with_FBA.R</td> <td>1-154</td> <td>numbers are in variable "model_explained"</td> </tr> <tr> <td>Fig. 4g (left)</td> <td>7_FBA_modeling/CR_model_final_run/2_DEG_modeling_supp_fused_with_FBA.R</td> <td>entire file</td> <td>corresponding figure produced by lines 342-345</td> </tr> <tr> <td>Fig. 4g (right)</td> <td>7_FBA_modeling/CR_model_final_run/2_DEG_modeling_supp_parameter_sensitivity_fused_with_FBA.R</td> <td>entire file</td> <td> </td> </tr> <tr> <td>Fig. 5a</td> <td>7_FBA_modeling/REVISION/human_perturb_seq/2_DEG_modeling_basic_CR_model.R</td> <td>1-297</td> <td> </td> </tr> <tr> <td>Fig. 5b</td> <td>7_FBA_modeling/REVISION/human_perturb_seq/3_DEG_modeling_edge_direction_test_randFBA.R</td> <td>entire file and see notes</td> <td>Reproducing this figure is complex because it integrates multiple randomization results; search "fitted_CR_interaction_diagram.pdf" to locate the code for making the figure, and track up for a few lines to load the dependency. To fully reproduce the analysis, follow the instructions in the Github and you will need to execute all two files named "3_DEG_modeling_edge_*.R".</td> </tr> <tr> <td>Fig. 5c</td> <td>7_FBA_modeling/REVISION/human_perturb_seq/2_DEG_modeling_supp_fused_with_FBA.R</td> <td>entire file</td> <td>corresponding figure produced by lines 343-346</td> </tr> <tr> <td>Fig. 5d</td> <td>7_FBA_modeling/REVISION/human_perturb_seq/2_DEG_modeling_supp_DEG_cutoff_sensitivity_fused_with_FBA.R</td> <td>entire file</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 1a</td> <td>1_QC_dataCleaning/5_supplementary_figure_rewiring.R</td> <td>193-219</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 1b</td> <td>1_QC_dataCleaning/5_supplementary_figure_rewiring.R</td> <td>1-91</td> <td>the published figure was remade manually for prettier representations, the plotting function for which is not provided in this code. However, the raw data stays the same.</td> </tr> <tr> <td>Extended Data Fig. 1c</td> <td>2_DE/SUPP_extra_figures_for_rewiring.R</td> <td>280-379</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 1d</td> <td>4_networkAnalysis/s2_rewiring_and_network_GPR.R</td> <td>387-555</td> <td>some inputs were generated by "4_networkAnalysis/s2_rewiring_and_network_GPR.m"</td> </tr> <tr> <td>Extended Data Fig. 1e</td> <td>4_networkAnalysis/s2_rewiring_and_network_GPR.R</td> <td>387-555</td> <td>some inputs were generated by "4_networkAnalysis/s2_rewiring_and_network_GPR.m"</td> </tr> <tr> <td>Extended Data Fig. 1f</td> <td>2_DE/4_2_pathway_level_analysis.R</td> <td>170-257</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 1g</td> <td>2_DE/4_2_pathway_level_analysis.R</td> <td>170-297</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 1h</td> <td>11_additional analysis/Wormsize/plot for publishing.R</td> <td>entire file</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 1i</td> <td>11_additional analysis/Wormsize/plot for publishing.R</td> <td>entire file</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 2a</td> <td>2_DE/3_1_DE_network_analysis.R</td> <td>1-61</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 2b</td> <td>2_DE/3_1_DE_network_analysis.R</td> <td>1-180</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 2c</td> <td>2_DE/3_1_DE_network_analysis.R</td> <td>1-152</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 2d</td> <td>2_DE/3_1_DE_network_analysis.R</td> <td>364-397</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 2e</td> <td>2_DE/3_1_DE_network_analysis.R</td> <td>364-383</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 2f</td> <td>2_DE/3_1_DE_network_analysis.R</td> <td>1-264</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 2g</td> <td>5_GSA/1_DE_annotation_pipeline.R</td> <td>1389-1522</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 3</td> <td>2_DE/5_1_1_DE_similarity_analysis.R</td> <td>468-765</td> <td>For producing the embedding, see "2_DE/clustering_python/cluster_RNAi_UMAP_DBSCAN.ipynb"</td> </tr> <tr> <td>Extended Data Fig. 4a</td> <td>4_networkAnalysis/s3_rewiring_and_network_connections.R</td> <td>1-254</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 4b</td> <td>4_networkAnalysis/s3_rewiring_and_network_connections.R</td> <td>258-303</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 4c</td> <td>2_DE/5_2_coexpression_analysis_direct_tree.R</td> <td>136-283</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 4d</td> <td>2_DE/SUPP_extra_figures_for_rewiring.R</td> <td>725-813</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 4e</td> <td>2_DE/5_2_coexpression_analysis_direct_tree.R</td> <td>136-589</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 4f</td> <td>11_additional analysis/regulators_nhr_sbp_tor_GEP_mGRN.R</td> <td>entire file</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 5a</td> <td>2_DE/6_3_scatter_plot_for_each_DEG_cluster_PCCtree.R</td> <td>1-224</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 5b</td> <td>2_DE/6_3_scatter_plot_for_each_DEG_cluster_PCCtree.R</td> <td>1-92</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 6a</td> <td>2_DE/6_1_biCluster_heatmaps_PCCtree.R (top row); 2_DE/6_2_rewiring_summary_map_PCCtree.R (bottom row)</td> <td>top row: 210-219; 229-238; bottom row: 1-162; 162-229</td> <td>These two scripts produced the last two figures in the first row and the first two figures in the second row. The remaining two figures are repetitive with other main/extended data figures, whose source code were specified there</td> </tr> <tr> <td>Extended Data Fig. 6b</td> <td>10_revision/2_tissue_expression_analysis.R</td> <td>1-184</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 6c</td> <td>10_revision/2_tissue_expression_analysis.R</td> <td>189-319</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 6d</td> <td>10_revision/2_tissue_expression_analysis.R</td> <td>326-480</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 7a-e</td> <td>2_DE/6_3_scatter_plot_for_each_DEG_cluster_PCCtree.R</td> <td>1-189</td> <td>The annotation bar plot is in 2_DE/SUPP_extra_figures_for_rewiring.R lines 394-715</td> </tr> <tr> <td>Extended Data Fig. 8a</td> <td>2_DE/3_1_DE_network_analysis.R</td> <td>649-753</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 8b</td> <td>2_DE/3_1_DE_network_analysis.R</td> <td>649-785</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 8c</td> <td>4_networkAnalysis/2_visualize_example_rewirings.R</td> <td>759-764</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 8d</td> <td>2_DE/4_2_pathway_level_analysis.R</td> <td>2247-2506</td> <td>the published figure uses level 4 and up direction (2_DE/figures/1_pathway_level_analysis/rxn_and_pathway_rewiring_LEVEL4_up.pdf)</td> </tr> <tr> <td>Extended Data Fig. 8e</td> <td>4_networkAnalysis/2_visualize_example_rewirings.R</td> <td>803-807</td> <td>please load the dependency starting from line 1</td> </tr> <tr> <td>Extended Data Fig. 8f</td> <td>4_networkAnalysis/2_visualize_example_rewirings.R</td> <td>870-874</td> <td>please load the dependency starting from line 1</td> </tr> <tr> <td>Extended Data Fig. 8g</td> <td>11_additional analysis/uPhe tracing/uphe RNAi.R</td> <td>entire file</td> <td>source data also provided along the paper</td> </tr> <tr> <td>Extended Data Fig. 9a</td> <td>4_networkAnalysis/2_visualize_example_rewirings.R</td> <td>1-601</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 9b</td> <td>4_networkAnalysis/2_visualize_example_rewirings.R</td> <td>691-714</td> <td>please load the dependency starting from line 1</td> </tr> <tr> <td>Extended Data Fig. 9c</td> <td>11_additional analysis/uGlucose tracing/uGlu rewiring.R</td> <td>entire file</td> <td>source data also provided along the paper</td> </tr> <tr> <td>Extended Data Fig. 9d</td> <td>4_networkAnalysis/2_visualize_example_rewirings.R</td> <td>607-629</td> <td>please load the dependency starting from line 1</td> </tr> <tr> <td>Extended Data Fig. 9e</td> <td>4_networkAnalysis/2_visualize_example_rewirings.R</td> <td>636-685</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 9f</td> <td>4_networkAnalysis/1_rewiring_network_modules.R</td> <td>entire file</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 10a</td> <td>2_DE/6_2_rewiring_summary_map_PCCtree.R</td> <td>entire file</td> <td>This figure is a subset of fig. 2f</td> </tr> <tr> <td>Extended Data Fig. 10c</td> <td>7_FBA_modeling/REVISION/1_cluster_CR_model.R</td> <td>entire file</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 11a</td> <td>7_FBA_modeling/CR_model_final_run/2_DEG_modeling_supp_excludeMultiObjGenes.R</td> <td>entire file</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 11b</td> <td>7_FBA_modeling/CR_model_final_run/2_DEG_modeling_supp_fused_with_FBA.R</td> <td>1-225</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 11c</td> <td>7_FBA_modeling/CR_model_final_run/2_DEG_modeling_supp_fused_with_FBA.R</td> <td>entire file</td> <td>corresponding figure produced by lines 337-340</td> </tr> <tr> <td>Extended Data Fig. 11d</td> <td>7_FBA_modeling/CR_model_final_run/2_DEG_modeling_supp_fused_with_FBA.R</td> <td>entire file</td> <td>corresponding figure produced by lines 352-355</td> </tr> <tr> <td>Extended Data Fig. 11e</td> <td>7_FBA_modeling/CR_model_final_run/2_DEG_modeling_supp_fused_with_FBA.R</td> <td>entire file</td> <td>corresponding figure produced by lines 362-365</td> </tr> <tr> <td>Extended Data Fig. 11f</td> <td>10_revision/2_tissue_expression_analysis.R</td> <td>486-742</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 12a</td> <td>7_FBA_modeling/REVISION/human_perturb_seq/2_DEG_modeling_basic_CR_model.R</td> <td>300-329</td> <td>run lines 1-101 to load dependency; Euler plot depends on a random seeds; we manually picked one output figure that has best visual representations; see the corresponding Github or "7_FBA_modeling/REVISION/human_perturb_seq/a1_gene_obj_classification.m" for generating the input matrix (core function matrix)</td> </tr> <tr> <td>Extended Data Fig. 12b</td> <td>7_FBA_modeling/REVISION/human_perturb_seq/1_EDA_of_human_perturb_seq.R</td> <td>1-176</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 12c</td> <td>7_FBA_modeling/REVISION/human_perturb_seq/2_DEG_modeling_basic_CR_model.R</td> <td>1-297</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 12d</td> <td>7_FBA_modeling/REVISION/human_perturb_seq/2_DEG_modeling_basic_CR_model.R</td> <td>1-297</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 12e</td> <td>7_FBA_modeling/REVISION/human_perturb_seq/2_DEG_modeling_supp_fused_with_FBA.R</td> <td>1-224</td> <td> </td> </tr> <tr> <td>Extended Data Fig. 12f</td> <td>7_FBA_modeling/REVISION/human_perturb_seq/2_DEG_modeling_supp_fused_with_FBA.R</td> <td>entire file</td> <td>corresponding figure produced by lines 337-341</td> </tr> <tr> <td>Extended Data Fig. 12g</td> <td>7_FBA_modeling/REVISION/human_perturb_seq/2_DEG_modeling_supp_fused_with_FBA.R</td> <td>entire file</td> <td>corresponding figure produced by lines 353-356</td> </tr> <tr> <td>Extended Data Fig. 12h</td> <td>7_FBA_modeling/REVISION/human_perturb_seq/2_DEG_modeling_supp_fused_with_FBA.R</td> <td>entire file</td> <td>corresponding figure produced by lines 363-366</td> </tr> <tr> <td>Extended Data Fig. 12i</td> <td>7_FBA_modeling/REVISION/human_perturb_seq/2_DEG_modeling_supp_parameter_sensitivity_fused_with_FBA.R</td> <td>entire file</td> <td> </td> </tr> <tr> <td>Supplementary Data Fig. 2a</td> <td>2_DE/5_1_1_DE_similarity_analysis.R</td> <td>467-572</td> <td> </td> </tr> <tr> <td>Supplementary Data Fig. 2b</td> <td>2_DE/6_4_Wald_heatmap_each_RNAi_Cluster.R</td> <td>entire file</td> <td> </td> </tr> <tr> <td>Supplementary Data Fig. 2c</td> <td>2_DE/5_1_2_DE_cluster_justification.R</td> <td>1-389</td> <td> </td> </tr> <tr> <td>Supplementary Data Fig. 2d</td> <td>2_DE/5_1_2_DE_cluster_justification.R</td> <td>1-308</td> <td> </td> </tr> <tr> <td>Supplementary Data Fig. 2e</td> <td>2_DE/5_1_2_DE_cluster_justification.R</td> <td>1-234</td> <td> </td> </tr> <tr> <td>Supplementary Data Fig. 2f</td> <td>12_FLEX_benchmark/run_FLEX_eval.sh</td> <td>entire file</td> <td>the bash scripts calls corresponding R scripts for reproducing the figures. You will need to install the FLEX package to run these codes. </td> </tr> <tr> <td>Supplementary Data Fig. 3, 4</td> <td>N/A</td> <td>N/A</td> <td>No coding involved as these figures were made manually</td> </tr> <tr> <td>Supplementary Data Fig. 5</td> <td>7_FBA_modeling/CR_model_final_run/2_DEG_modeling_basic_CR_model.R</td> <td>1-270</td> <td> </td> </tr> <tr> <td>Supplementary Data Fig. 6</td> <td>7_FBA_modeling/CR_model_final_run/2_DEG_modeling_basic_CR_model.R</td> <td>274-578</td> <td>please load the dependency starting from line 1</td> </tr> </tbody> </table> a: The lines indicate the chunk of codes to reproduce the corresponding figure; The figure is reproduced at the end of the referred codes. Please note that you may have to run the codes above the referred chunk (i.e., from the first line) to load dependent variables to execute the referred codes. However, you should be able to reproduce the figure only with the codes within the referred script.  Notice: We advise you download the entire project working directory (including all zip files and unzip them into corresponding folders), for reproducing any analysis. If you only download the zip file relevant to your figure of interest, you may or may not run into issues due to the missing files in another folder.   Contact: For any questions, please contact Xuhang (Hang) Li at Xuhang.Li@umassmed.edu. (we plan to deposit an updated version with better code annotations, but cannot complete that currently due to time restirctions)

ShareScore

44/100

Overall dataset sharing score

Score breakdown

These five areas show where the dataset supports — or may limit — practical reuse.

Stewardship: 8
Harmonization: 8
Access: 16
Reuse readiness: 8
Engagement: 4

Topics

metabolic network transcriptional rewiring metabolic rewiring transcriptional regulation