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-69278     R-HSA-69278
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
                                                                                       Description
R-HSA-69278                                                                    Cell Cycle, Mitotic
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
              GeneRatio   BgRatio RichFactor FoldEnrichment   zScore
R-HSA-69278      49/314 479/10173  0.1022965       3.314210 9.259137
R-HSA-69620      33/314 244/10173  0.1352459       4.381709 9.541792
R-HSA-69618      19/314  94/10173  0.2021277       6.548550 9.644646
R-HSA-2500257    20/314 109/10173  0.1834862       5.944604 9.262132
R-HSA-141424     17/314  79/10173  0.2151899       6.971741 9.508990
R-HSA-141444     17/314  79/10173  0.2151899       6.971741 9.508990
                    pvalue     p.adjust       qvalue
R-HSA-69278   6.635185e-14 5.288243e-11 4.742411e-11
R-HSA-69620   5.785253e-13 2.305423e-10 2.067467e-10
R-HSA-69618   5.135524e-11 1.364337e-08 1.223516e-08
R-HSA-2500257 1.006794e-10 2.006036e-08 1.798981e-08
R-HSA-141424  1.923239e-10 2.554702e-08 2.291016e-08
R-HSA-141444  1.923239e-10 2.554702e-08 2.291016e-08
                                                                                                                                                                                                                                                                                                   geneID
R-HSA-69278   CDC45/CDCA8/CDC20/FOXM1/CENPE/MYBL2/CCNB2/NDC80/TOP2A/NCAPH/RRM2/UBE2C/HJURP/SKA1/TPX2/NEK2/CENPM/CENPN/UBE2S/CCNA2/CDK1/ERCC6L/GINS1/KIF18A/CDT1/NCAPG/AURKB/GINS2/KIF20A/AURKA/CCNB1/MCM5/PTTG1/MCM2/KIF2C/CDC25A/CDC6/PLK1/BUB1B/GTSE1/ZWINT/SPC25/CENPI/ESPL1/TK1/CCNE1/ORC6/ORC1/TAOK1
R-HSA-69620                                                                                              CDC45/CDCA8/CDC20/CENPE/CCNB2/NDC80/UBE2C/SKA1/CENPM/CENPN/UBE2S/CCNA2/CDK1/ERCC6L/KIF18A/AURKB/CHEK1/CCNB1/MCM5/MCM2/KIF2C/CDC25A/CDC6/PLK1/BUB1B/GTSE1/ZWINT/SPC25/CENPI/CCNE1/ORC6/ORC1/TAOK1
R-HSA-69618                                                                                                                                                                             CDCA8/CDC20/CENPE/NDC80/UBE2C/SKA1/CENPM/CENPN/UBE2S/ERCC6L/KIF18A/AURKB/KIF2C/PLK1/BUB1B/ZWINT/SPC25/CENPI/TAOK1
R-HSA-2500257                                                                                                                                                                      CDCA8/CDC20/CENPE/CCNB2/NDC80/SKA1/CENPM/CENPN/CDK1/ERCC6L/KIF18A/AURKB/CCNB1/KIF2C/PLK1/BUB1B/ZWINT/SPC25/CENPI/TAOK1
R-HSA-141424                                                                                                                                                                                        CDCA8/CDC20/CENPE/NDC80/SKA1/CENPM/CENPN/ERCC6L/KIF18A/AURKB/KIF2C/PLK1/BUB1B/ZWINT/SPC25/CENPI/TAOK1
R-HSA-141444                                                                                                                                                                                        CDCA8/CDC20/CENPE/NDC80/SKA1/CENPM/CENPN/ERCC6L/KIF18A/AURKB/KIF2C/PLK1/BUB1B/ZWINT/SPC25/CENPI/TAOK1
              Count
R-HSA-69278      49
R-HSA-69620      33
R-HSA-69618      19
R-HSA-2500257    20
R-HSA-141424     17
R-HSA-141444     17

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-69278     R-HSA-69278                  Cell Cycle, Mitotic     382
R-HSA-453279   R-HSA-453279 Mitotic G1 phase and G1/S transition     113
R-HSA-69620     R-HSA-69620               Cell Cycle Checkpoints     194
R-HSA-2555396 R-HSA-2555396       Mitotic Metaphase and Anaphase     156
R-HSA-69206     R-HSA-69206                      G1/S Transition     100
R-HSA-68882     R-HSA-68882                     Mitotic Anaphase     155
              enrichmentScore      NES pvalue     p.adjust       qvalue rank
R-HSA-69278         0.6235874 3.027524  1e-10 4.463636e-09 3.240829e-09 1769
R-HSA-453279        0.7235270 2.995080  1e-10 4.463636e-09 3.240829e-09 1155
R-HSA-69620         0.6662086 2.973969  1e-10 4.463636e-09 3.240829e-09 1768
R-HSA-2555396       0.6639936 2.919566  1e-10 4.463636e-09 3.240829e-09 1768
R-HSA-69206         0.7204590 2.917355  1e-10 4.463636e-09 3.240829e-09 1155
R-HSA-68882         0.6619226 2.906883  1e-10 4.463636e-09 3.240829e-09 1768
                                leading_edge
R-HSA-69278   tags=38%, list=14%, signal=34%
R-HSA-453279   tags=44%, list=9%, signal=41%
R-HSA-69620   tags=42%, list=14%, signal=36%
R-HSA-2555396 tags=42%, list=14%, signal=36%
R-HSA-69206    tags=45%, list=9%, signal=41%
R-HSA-68882   tags=41%, list=14%, signal=36%
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 core_enrichment
R-HSA-69278   8318/55143/991/2305/1062/4605/9133/10403/7153/23397/6241/11065/55355/220134/22974/4751/79019/55839/27338/890/983/54821/9837/81930/81620/64151/9212/51659/10112/6790/891/4174/9232/4171/11004/993/990/5347/701/51512/11130/57405/2491/9700/7083/898/23594/4998/10733/1163/54801/9134/4175/4173/2237/5984/994/9918/7298/84296/699/4609/1063/85236/5111/28227/9688/64785/5709/26271/55055/51053/1869/1029/1763/1719/8970/54892/8357/5693/8317/7277/5427/23649/7112/5713/3930/79980/9735/55732/11047/5557/23310/5691/1871/1031/80086/995/10051/1104/5685/84790/1019/5558/4172/9631/5424/79866/5690/5885/5686/5695/203068/7027/10213/1875/23175/8345/7283/10381/5983/7979/54908/10592/23165/10383/3018/22822/5714/10376/7443/55706/8815/5702/3014/1030/23636/9978/5905/5425/5708/55166/5692/10393/54107
R-HSA-453279                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         8318/4605/7153/6241/890/983/81620/891/4174/4171/993/990/7083/898/23594/4998/1163/9134/4175/4173/7298/4609/5111/28227/5709/26271/51053/1869/1029/1719/5693/8317/5427/23649/5713/11047/5557/5691/1871/1031/5685/1019/5558/4172/5690/5686/5695/7027/10213/1875
R-HSA-69620                                                                                                                                                                                                                                                                                                                                                                 8318/55143/991/1062/9133/10403/11065/220134/79019/55839/27338/890/983/54821/81930/9212/1111/891/4174/4171/11004/993/990/5347/701/51512/11130/57405/2491/898/23594/4998/9134/4175/4173/5984/699/1063/85236/5709/55055/641/1029/1763/8970/5693/8317/5713/79980/9735/11047/2810/5691/995/5685/4172/7336/5690/83990/5686/5695/11200/10213/8345/7534/80010/5983/7979/54908/4683/63967/3018/5714/5702/3014/9978/5905/5708/55166/5692/10393
R-HSA-2555396                                                                                                                                                                                                                                                                                                                                                                                                                                       55143/991/1062/9133/10403/11065/220134/79019/55839/27338/983/54821/81930/9212/891/9232/11004/5347/701/11130/57405/2491/9700/699/1063/9688/5709/26271/55055/5693/7277/7112/5713/3930/79980/9735/11047/5691/80086/1104/5685/84790/9631/5690/5885/5686/5695/10213/10381/7979/54908/23165/10383/5714/10376/7443/55706/8815/5702/23636/5905/5708/55166/5692/10393
R-HSA-69206                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   8318/6241/890/983/81620/891/4174/4171/993/990/7083/898/23594/4998/1163/9134/4175/4173/7298/4609/5111/28227/5709/26271/51053/1869/1719/5693/8317/5427/23649/5713/11047/5557/5691/5685/1019/5558/4172/5690/5686/5695/7027/10213/1875
R-HSA-68882                                                                                                                                                                                                                                                                                                                                                                                                                                               55143/991/1062/9133/10403/11065/220134/79019/55839/27338/983/54821/81930/9212/891/9232/11004/5347/701/11130/57405/2491/9700/699/1063/9688/5709/55055/5693/7277/7112/5713/3930/79980/9735/11047/5691/80086/1104/5685/84790/9631/5690/5885/5686/5695/10213/10381/7979/54908/23165/10383/5714/10376/7443/55706/8815/5702/23636/5905/5708/55166/5692/10393

7.4 Pathway Visualization

ReactomePA implemented the viewPathway() to visualized selected reactome pathway. More general purpose of visualization methods for ORA and GSEA results are provided in the enrichplot package and are documented on Chapter 14.

(ref:viewpathwayscap) Visualize reactome pathway.

(ref:viewpathwaycap) Visualize reactome pathway.

viewPathway("E2F mediated regulation of DNA replication", 
            readable = TRUE, 
            foldChange = geneList)

(ref:viewpathwaycap)

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.