7  Reactome enrichment analysis

ReactomePA is designed for reactome pathway based analysis (Yu and He 2016). Reactome is an open-source, open access, manually curated and peer-reviewed pathway database.

7.1 Supported organisms

Currently ReactomePA supports several model organisms, including ‘celegans’, ‘fly’, ‘human’, ‘mouse’, ‘rat’, ‘yeast’ and ‘zebrafish’. The input gene ID should be Entrez gene ID. We recommend using clusterProfiler::bitr() to convert biological IDs.

7.2 Reactome pathway over-representation analysis

Enrichment analysis is a widely used approach to identify biological themes. ReactomePA implemented enrichPathway() that uses hypergeometric model to assess whether the number of selected genes associated with a reactome pathway is larger than expected.

library(ReactomePA)
data(geneList, package="DOSE"
)
de <- names(geneList)[abs(geneList) > 1.5]
head(de)
[1] "4312"  "8318"  "10874" "55143" "55388" "991"  
x <- enrichPathway(gene=de, pvalueCutoff = 0.05, readable=TRUE)
head(x)
                         ID
R-HSA-69620     R-HSA-69620
R-HSA-69618     R-HSA-69618
R-HSA-2500257 R-HSA-2500257
R-HSA-141424   R-HSA-141424
R-HSA-141444   R-HSA-141444
R-HSA-68882     R-HSA-68882
                                                                                       Description
R-HSA-69620                                                                 Cell Cycle Checkpoints
R-HSA-69618                                                             Mitotic Spindle Checkpoint
R-HSA-2500257                                              Resolution of Sister Chromatid Cohesion
R-HSA-141424                                         Amplification of signal from the kinetochores
R-HSA-141444  Amplification  of signal from unattached  kinetochores via a MAD2  inhibitory signal
R-HSA-68882                                                                       Mitotic Anaphase
              GeneRatio   BgRatio RichFactor FoldEnrichment    zScore
R-HSA-69620      38/342 292/11287  0.1301370       4.294901 10.083502
R-HSA-69618      22/342 113/11287  0.1946903       6.425348 10.245607
R-HSA-2500257    23/342 129/11287  0.1782946       5.884242  9.862206
R-HSA-141424     20/342  96/11287  0.2083333       6.875609 10.219491
R-HSA-141444     20/342  96/11287  0.2083333       6.875609 10.219491
R-HSA-68882      28/342 225/11287  0.1244444       4.107031  8.321376
                    pvalue     p.adjust       qvalue
R-HSA-69620   1.988463e-14 3.428110e-11 3.428110e-11
R-HSA-69618   2.386464e-12 2.057132e-09 2.057132e-09
R-HSA-2500257 5.135365e-12 2.245208e-09 2.245208e-09
R-HSA-141424  6.511625e-12 2.245208e-09 2.245208e-09
R-HSA-141444  6.511625e-12 2.245208e-09 2.245208e-09
R-HSA-68882   1.865374e-10 5.101358e-08 5.101358e-08
                                                                                                                                                                                                                                      geneID
R-HSA-69620   CDK1/EXO1/ERCC6L/MCM5/SKA1/CDC25A/UBE2S/CDC45/CDC6/CENPN/CDC20/SPC25/BUB1B/ORC1/CCNA2/MCM2/KIF18A/AURKB/UBE2C/CCNE1/CENPE/TAOK1/GTSE1/CHEK1/BIRC5/CENPI/ZWINT/CDCA8/CCNB2/MCM10/PLK1/CENPM/NDC80/ORC6/MAD2L1/CENPU/KIF2C/CCNB1
R-HSA-69618                                                                                             ERCC6L/SKA1/UBE2S/CENPN/CDC20/SPC25/BUB1B/KIF18A/AURKB/UBE2C/CENPE/TAOK1/BIRC5/CENPI/ZWINT/CDCA8/PLK1/CENPM/NDC80/MAD2L1/CENPU/KIF2C
R-HSA-2500257                                                                                      CDK1/ERCC6L/SKA1/CENPN/CDC20/SPC25/BUB1B/KIF18A/AURKB/CENPE/TAOK1/BIRC5/CENPI/ZWINT/CDCA8/CCNB2/PLK1/CENPM/NDC80/MAD2L1/CENPU/KIF2C/CCNB1
R-HSA-141424                                                                                                        ERCC6L/SKA1/CENPN/CDC20/SPC25/BUB1B/KIF18A/AURKB/CENPE/TAOK1/BIRC5/CENPI/ZWINT/CDCA8/PLK1/CENPM/NDC80/MAD2L1/CENPU/KIF2C
R-HSA-141444                                                                                                        ERCC6L/SKA1/CENPN/CDC20/SPC25/BUB1B/KIF18A/AURKB/CENPE/TAOK1/BIRC5/CENPI/ZWINT/CDCA8/PLK1/CENPM/NDC80/MAD2L1/CENPU/KIF2C
R-HSA-68882                                                          CDK1/ERCC6L/SKA1/LMNB1/UBE2S/CENPN/CDC20/SPC25/BUB1B/PTTG1/KIF18A/AURKB/UBE2C/CENPE/TAOK1/BIRC5/ESPL1/CENPI/ZWINT/CDCA8/CCNB2/PLK1/CENPM/NDC80/MAD2L1/CENPU/KIF2C/CCNB1
              Count
R-HSA-69620      38
R-HSA-69618      22
R-HSA-2500257    23
R-HSA-141424     20
R-HSA-141444     20
R-HSA-68882      28

7.3 Reactome pathway gene set enrichment analysis

y <- gsePathway(geneList, 
                pvalueCutoff = 0.2,
                pAdjustMethod = "BH", 
                verbose = FALSE)
head(y)
                       ID                          Description setSize
R-HSA-453279 R-HSA-453279 Mitotic G1 phase and G1/S transition     132
R-HSA-69620   R-HSA-69620               Cell Cycle Checkpoints     238
R-HSA-69206   R-HSA-69206                      G1/S Transition     114
R-HSA-69239   R-HSA-69239                     Synthesis of DNA      99
R-HSA-69481   R-HSA-69481                     G2/M Checkpoints     125
R-HSA-69306   R-HSA-69306                      DNA Replication     133
             enrichmentScore      NES pvalue     p.adjust       qvalue rank
R-HSA-453279       0.7117138 2.998834  1e-10 5.070588e-09 5.070588e-09 1155
R-HSA-69620        0.6508575 2.994081  1e-10 5.070588e-09 5.070588e-09 1905
R-HSA-69206        0.7163844 2.951798  1e-10 5.070588e-09 5.070588e-09 1155
R-HSA-69239        0.7192759 2.938992  1e-10 5.070588e-09 5.070588e-09 1905
R-HSA-69481        0.6968898 2.930468  1e-10 5.070588e-09 5.070588e-09 1905
R-HSA-69306        0.6877315 2.927944  1e-10 5.070588e-09 5.070588e-09 1769
                               leading_edge
R-HSA-453279  tags=43%, list=9%, signal=40%
R-HSA-69620  tags=42%, list=15%, signal=36%
R-HSA-69206   tags=45%, list=9%, signal=41%
R-HSA-69239  tags=57%, list=15%, signal=48%
R-HSA-69481  tags=50%, list=15%, signal=43%
R-HSA-69306  tags=47%, list=14%, signal=41%
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  core_enrichment
R-HSA-453279                                                                                                                                                                                                                                     8318/55388/4605/7153/6241/890/983/81620/891/4174/4171/993/990/7083/898/23594/4998/1163/9134/4175/4173/10926/6502/7298/4609/5111/28227/5688/5709/26271/51053/1869/1029/1719/4067/5693/8317/5427/23649/4176/5713/11047/5557/5691/1871/1031/5685/1019/5558/4172/5690/5684/5686/5695/7027/10213/1875
R-HSA-69620  8318/55143/55388/991/1062/9133/10403/11065/220134/79019/55839/27338/890/983/54821/4085/81930/332/9212/1111/891/4174/4171/11004/993/990/5347/701/51512/9156/11130/79682/57405/2491/898/23594/4998/9134/4175/4173/10926/5984/1058/699/1063/85236/5688/5709/55055/641/1029/1763/8970/5693/8317/4176/5713/79980/5982/9735/11047/2810/5691/9088/995/5685/7468/4172/7336/5690/5684/83990/5686/5695/11200/10213/8345/7534/80010/5983/7979/54908/6396/4683/63967/3018/5714/5702/3014/9978/5905/3619/5708/55166/5692/10393/8290/5704/580/6119
R-HSA-69206                                                                                                                                                                                                                                                                    8318/55388/6241/890/983/81620/891/4174/4171/993/990/7083/898/23594/4998/1163/9134/4175/4173/10926/6502/7298/4609/5111/28227/5688/5709/26271/51053/1869/1719/5693/8317/5427/23649/4176/5713/11047/5557/5691/5685/1019/5558/4172/5690/5684/5686/5695/7027/10213/1875
R-HSA-69239                                                                                                                                                                                                                                     8318/11065/27338/890/9837/81620/51659/4174/4171/990/898/23594/4998/9134/4175/4173/2237/6502/5984/84296/5111/5688/64785/5709/51053/1763/5693/5427/23649/4176/5713/5982/11047/5557/5691/5685/5558/4172/5424/5690/5684/5686/5695/10213/5983/7979/5714/5702/9978/5425/5708/5692/10393/54107/5704/6119
R-HSA-69481                                                                                                                                                                                                         8318/55388/9133/890/983/1111/891/4174/4171/993/990/51512/9156/23594/4998/4175/4173/10926/5984/85236/5688/5709/641/1763/8970/5693/8317/4176/5713/5982/11047/2810/5691/9088/995/5685/7468/4172/7336/5690/5684/83990/5686/5695/11200/10213/8345/7534/80010/5983/7979/4683/63967/3018/5714/5702/3014/5708/5692/8290/5704/580/6119
R-HSA-69306                                                                                                                                                                                               8318/55388/11065/27338/890/9837/81620/51659/4174/4171/990/898/23594/4998/9134/4175/4173/2237/10926/6502/5984/84296/85236/5111/5688/64785/5709/51053/1763/8970/8357/5693/8317/5427/23649/4176/5713/5982/11047/5557/5691/5685/5558/4172/5424/5690/5684/5686/5695/10213/8345/5983/7979/3018/5714/5702/3014/9978/5425/5708/5692/10393/54107
             log2err
R-HSA-453279     NaN
R-HSA-69620      NaN
R-HSA-69206      NaN
R-HSA-69239      NaN
R-HSA-69481      NaN
R-HSA-69306      NaN

7.4 Pathway Visualization

ReactomePA implemented the viewPathway() to visualize selected reactome pathways. More general purpose visualization methods for ORA and GSEA results are provided in the enrichplot package and are documented in Chapter 13. See Figure 7.1 for an example.

viewPathway("E2F mediated regulation of DNA replication", 
            readable = TRUE, 
            foldChange = geneList)
Figure 7.1: Visualize reactome pathway.

References

Yu, Guangchuang, and Qing-Yu He. 2016. ReactomePA: An r/Bioconductor Package for Reactome Pathway Analysis and Visualization.” Molecular BioSystems 12 (2): 477–79. https://doi.org/10.1039/C5MB00663E.