Func6onal Profiling from Babelomics 5

Funconal proﬁling from Babelomics 5

Dan Crespo [email protected] March 3th, 2016 Where are we?

General NGS pipeline: Funconal analysis: • Single enrichment: FaGO • Gene set enrichment: Logisc Model

Genomics Transcriptomics Where are we?

Funconal analysis Implemented in:

Diﬀerenal expression hp://babelomics.org

Genes Gene Sets

FaGO GSA logisc model Quesons we try to answer

• Is there any signiﬁcant funconal enrichment in my gene list / gene sets? Quesons we try to answer

• Is there any signiﬁcat funconal enrichment in my gene list / gene sets? • Are these genes involved in common pathways? Quesons we try to answer

• Is there any signiﬁcat funconal enrichment in my gene list / gene sets? • Are these genes involved in common pathways? • Do they share speciﬁc regulaon? Quesons we try to answer

• Is there any signiﬁcat funconal enrichment in my gene list / gene sets? • Are these genes involved in common pathways? • Do they share speciﬁc regulaon? • Are they involved in the same disease? Babelomics 5: FaGO Single enrichment

Comparison between two list of genes with user deﬁned annotaons Single enrichment

Comparison between two list of genes with user deﬁned annotaons

List 1: User provided List 2: Funconal annotaons: • Gene symbols • User provided • Pre-deﬁned annotaons • Probe Ids • Rest of genome • Custom annotaons • Entrez Ids … • Complementary list

Mulple tesng context Fisher exact test

Benjamini – Hochberg p-value correcon

Ranked results Single enrichment

List 1 List 2 Funconal annotaons AATF BIRC7 CIAS1 PEA15 ACTA1 DNAI1 KIF3B MYO15A Apoptoc process GO:0006915 APAF1 BIRC8 CIAS1 PNUTL2 ACTA2 DNAI2 KIF3C MYO15B API5 BNIP1 CIDEA PRKAA1 ACTB DNAI2 KIF4A MYO18B AVEN BNIP2 CIDEA RIPK2 ACTC DNAL4 KIF5A MYO1A BAG1 BNIP3 CIDEB RTN4 ACTG1 DNALI1 KIF5B MYO1A CBX4 BRCA1 AVEN MCL1 BAG2 BOK CRADD SON ACTG2 DNCH1 KIF5B MYO1B CRADD HDAC3 FAIM2 BCL2L2 BAG3 BRCA1 CRADD SPHK2 ACTL6 DNCI1 KIF5C MYO1C IL10 HDAC1 BID BCL2L1 BAG4 CARD10 DAD1 SPP1 ACTR1A DNCI2 KIF9 MYO1D BAK1 RTN4 TNFRSF25 PEA15 BAG5 CARD11 DEDD SPP1 ACTR1B DNCL1 KIFC1 MYO1E BAK1 CARD12 DFFA TNFAIP3 APPBP2 DNCL2A KIFC3 MYO1E BIRC8 IER3 BCL2 CFLAR BAX CARD12 DFFB TNFRSF10A ATSV DNCL2B KNS2 MYO1F API5 HTATIP2 DAD1 CARD9 C20orf23 DNCLI2 KNSL7 MYO1G BCL10 CARD14 EBAG9 TNFRSF10B BCL10 DEDD PRKAA1 SPHK2 BCL2 CARD15 EBAG9 TNFRSF25 CENPE DNM1 MPHOSPH1 MYO3A BCL2 CARD4 FADD TNFRSF6 DCTN1 DNM2 MYH1 MYO3B OPA1 BNIP3 ACTC CIDEB BCL2A1 CARD4 FAIM2 TNFRSF6B DCTN2 KIF11 MYH1 MYO3B DFFA PDCD2 BIRC6 CASP2 BCL2L1 CARD6 HDAC1 DNAH1 KIF13A MYH10 MYO5A DFFB CASP5 BIRC5 EBAG9 DNAH11 KIF13B MYH11 MYO5B BCL2L10 CARD9 HDAC3 BCL2A1 BAG5 BIRC3 NOL3 BCL2L13 CASP1 HRK DNAH12 KIF14 MYH13 MYO5C BCL2L2 CASP10 HTATIP2 DNAH14 KIF17 MYH13 MYO6 HRK BAG4 BIRC2 CARD10 BCL2L7P1 CASP10 IER3 DNAH14 KIF1B MYH2 MYO6 FADD CASP4 BIRC1 TNFRSF10A DNAH14 KIF1C MYH2 MYO7A BFAR CASP10 IER3 DNAH14 KIF2 MYH3 MYO7B BCL2L13 CASP9 CASP10 CARD11 BID CASP2 IER3 DNAH14 KIF20A MYH4 MYO9A BCL2L10 BAG1 SON CARD14 BIK CASP2 IL10 DNAH17 KIF22 MYH4 MYO9B BIRC1 CASP2 IL18 NME6 BOK BAX KIF1B DNAH2 KIF22 MYH6 OPA1 BIRC1 CASP4 IL2 RIPK2 BAG3 BIK DNM2 DNAH3 KIF23 MYH6 RPLP0P1 BIRC2 CASP5 MALT1 DNAH5 KIF23 MYH7 RPLP0P1 TNFAIP3 CASP8 APAF1 TNFRSF10B BIRC3 CASP8 MCL1 DNAH7 KIF25 MYH7B TCTE1 TNFRSF6B CASP1 TIAF1 BNIP1 BIRC4 CASP9 NME6 DNAH8 KIF26A MYH8 TCTEL1 BNIP2 CARD6 MALT1 CIDEA BIRC5 CBX4 NOL3 DNAH9 KIF2C MYH9 TIAF1 BIRC6 CFLAR PDCD2 DNAH9 KIF3A MYO10 Single enrichment

List 1 List 2 Funconal annotaons AATF BIRC7 CIAS1 PEA15 ACTA1 DNAI1 KIF3B MYO15A Posive regulaon of immune APAF1 BIRC8 CIAS1 PNUTL2 ACTA2 DNAI2 KIF3C MYO15B API5 BNIP1 CIDEA PRKAA1 ACTB DNAI2 KIF4A MYO18B system process GO0002684 AVEN BNIP2 CIDEA RIPK2 ACTC DNAL4 KIF5A MYO1A BAG1 BNIP3 CIDEB RTN4 ACTG1 DNALI1 KIF5B MYO1A IL18 RIPK2 MALT1 ACTB BAG2 BOK CRADD SON ACTG2 DNCH1 KIF5B MYO1B ACTG1 TNFAIP3 MYO10 BIRC3 BAG3 BRCA1 CRADD SPHK2 ACTL6 DNCI1 KIF5C MYO1C BCL10 CARD9 CASP8 BIRC2 BAG4 CARD10 DAD1 SPP1 ACTR1A DNCI2 KIF9 MYO1D MYH2 MYO1C CARD11 BAX BAG5 CARD11 DEDD SPP1 ACTR1B DNCL1 KIFC1 MYO1E BAK1 CARD12 DFFA TNFAIP3 APPBP2 DNCL2A KIFC3 MYO1E FADD MYO1G BCL2 IL2 BAX CARD12 DFFB TNFRSF10A ATSV DNCL2B KNS2 MYO1F BCL10 CARD14 EBAG9 TNFRSF10B C20orf23 DNCLI2 KNSL7 MYO1G BCL2 CARD15 EBAG9 TNFRSF25 CENPE DNM1 MPHOSPH1 MYO3A BCL2 CARD4 FADD TNFRSF6 DCTN1 DNM2 MYH1 MYO3B BCL2A1 CARD4 FAIM2 TNFRSF6B DCTN2 KIF11 MYH1 MYO3B BCL2L1 CARD6 HDAC1 DNAH1 KIF13A MYH10 MYO5A BCL2L10 CARD9 HDAC3 DNAH11 KIF13B MYH11 MYO5B BCL2L13 CASP1 HRK DNAH12 KIF14 MYH13 MYO5C BCL2L2 CASP10 HTATIP2 DNAH14 KIF17 MYH13 MYO6 BCL2L7P1 CASP10 IER3 DNAH14 KIF1B MYH2 MYO6 DNAH14 KIF1C MYH2 MYO7A BFAR CASP10 IER3 DNAH14 KIF2 MYH3 MYO7B BID CASP2 IER3 DNAH14 KIF20A MYH4 MYO9A BIK CASP2 IL10 DNAH17 KIF22 MYH4 MYO9B BIRC1 CASP2 IL18 DNAH2 KIF22 MYH6 OPA1 BIRC1 CASP4 IL2 DNAH3 KIF23 MYH6 OPA1 BIRC2 CASP5 MALT1 DNAH5 KIF23 MYH7 RPLP0P1 BIRC3 CASP8 MCL1 DNAH7 KIF25 MYH7B RPLP0P1 BIRC4 CASP9 NME6 DNAH8 KIF26A MYH8 TCTE1 BIRC5 CBX4 NOL3 DNAH9 KIF2C MYH9 TCTEL1 BIRC6 CFLAR PDCD2 DNAH9 KIF3A MYO10 TIAF1 Single enrichment

List 1 List 2 Funconal annotaons AATF BIRC7 CIAS1 PEA15 ACTA1 DNAI1 KIF3B MYO15A Posive regulaon of immune APAF1 BIRC8 CIAS1 PNUTL2 ACTA2 DNAI2 KIF3C MYO15B API5 BNIP1 CIDEA PRKAA1 ACTB DNAI2 KIF4A MYO18B system process GO0002684 AVEN BNIP2 CIDEA RIPK2 ACTC DNAL4 KIF5A MYO1A BAG1 BNIP3 CIDEB RTN4 ACTG1 DNALI1 KIF5B MYO1A IL18 RIPK2 MALT1 ACTB BAG2 BOK CRADD SON ACTG2 DNCH1 KIF5B MYO1B ACTG1 TNFAIP3 MYO10 BIRC3 BAG3 BRCA1 CRADD SPHK2 ACTL6 DNCI1 KIF5C MYO1C BCL10 CARD9 CASP8 BIRC2 BAG4 CARD10 DAD1 SPP1 ACTR1A DNCI2 KIF9 MYO1D MYH2 MYO1C CARD11 BAX BAG5 CARD11 DEDD SPP1 ACTR1B DNCL1 KIFC1 MYO1E BAK1 CARD12 DFFA TNFAIP3 APPBP2 DNCL2A KIFC3 MYO1E FADD MYO1G BCL2 IL2 BAX CARD12 DFFB TNFRSF10A ATSV DNCL2B KNS2 MYO1F BCL10 CARD14 EBAG9 TNFRSF10B C20orf23 DNCLI2 KNSL7 MYO1G Organ development GO0048515 BCL2 CARD15 EBAG9 TNFRSF25 CENPE DNM1 MPHOSPH1 MYO3A BCL2 CARD4 FADD TNFRSF6 DCTN1 DNM2 MYH1 MYO3B IL18 MYH6 KIF3A MYO7A BCL2A1 CARD4 FAIM2 TNFRSF6B DCTN2 KIF11 MYH1 MYO3B IL10 MYH9 SPHK2 CASP5 BCL2L1 CARD6 HDAC1 DNAH1 KIF13A MYH10 MYO5A BAK1 MYO18B MYO1E CASP9 DNAH11 KIF13B MYH11 MYO5B BCL2L10 CARD9 HDAC3 MYO6 RIPK2 MALT1 BOK BCL2L13 CASP1 HRK DNAH12 KIF14 MYH13 MYO5C BCL2L2 CASP10 HTATIP2 DNAH14 KIF17 MYH13 MYO6 MYO3A BCL2L2 HDAC1 BAG3 BCL2L7P1 CASP10 IER3 DNAH14 KIF1B MYH2 MYO6 MYH3 BCL2L1 MYH11 CASP8 DNAH14 KIF1C MYH2 MYO7A BFAR CASP10 IER3 DFFB MYO15A MYH10 CASP2 DNAH14 KIF2 MYH3 MYO7B BID CASP2 IER3 DNAH14 KIF20A MYH4 MYO9A MYH7 SPP1 RTN4 ACTA1 BIK CASP2 IL10 DNAH17 KIF22 MYH4 MYO9B BIRC1 CASP2 IL18 FADD CFLAR DEDD ACTA2 DNAH2 KIF22 MYH6 OPA1 BIRC1 CASP4 IL2 ACTB DNAH5 MYO5A CARD11 DNAH3 KIF23 MYH6 OPA1 BIRC2 CASP5 MALT1 DNAH5 KIF23 MYH7 RPLP0P1 ACTC BCL2 BID FAIM2 BIRC3 CASP8 MCL1 DNAH7 KIF25 MYH7B RPLP0P1 BIRC6 BAX BIK APAF1 BIRC4 CASP9 NME6 DNAH8 KIF26A MYH8 TCTE1 IL2 BIRC5 CBX4 NOL3 DNAH9 KIF2C MYH9 TCTEL1 BIRC6 CFLAR PDCD2 DNAH9 KIF3A MYO10 TIAF1 Single enrichment

List 1 List 2 Funconal annotaons AATF BIRC7 CIAS1 PEA15 ACTA1 DNAI1 KIF3B MYO15A Posive regulaon of immune APAF1 BIRC8 CIAS1 PNUTL2 ACTA2 DNAI2 KIF3C MYO15B API5 BNIP1 CIDEA PRKAA1 ACTB DNAI2 KIF4A MYO18B system process GO0002684 AVEN BNIP2 CIDEA RIPK2 ACTC DNAL4 KIF5A MYO1A BAG1 BNIP3 CIDEB RTN4 ACTG1 DNALI1 KIF5B MYO1A IL18 RIPK2 MALT1 ACTB BAG2 BOK CRADD SON ACTG2 DNCH1 KIF5B MYO1B ACTG1 TNFAIP3 MYO10 BIRC3 BAG3 BRCA1 CRADD SPHK2 ACTL6 DNCI1 KIF5C MYO1C BCL10 CARD9 CASP8 BIRC2 BAG4 CARD10 DAD1 SPP1 ACTR1A DNCI2 KIF9 MYO1D MYH2 MYO1C CARD11 BAX BAG5 CARD11 DEDD SPP1 ACTR1B DNCL1 KIFC1 MYO1E BAK1 CARD12 DFFA TNFAIP3 APPBP2 DNCL2A KIFC3 MYO1E FADD MYO1G BCL2 IL2 BAX CARD12 DFFB TNFRSF10A ATSV DNCL2B KNS2 MYO1F BCL10 CARD14 EBAG9 TNFRSF10B C20orf23 DNCLI2 KNSL7 MYO1G Organ development GO0048515 BCL2 CARD15 EBAG9 TNFRSF25 CENPE DNM1 MPHOSPH1 MYO3A BCL2 CARD4 FADD TNFRSF6 DCTN1 DNM2 MYH1 MYO3B IL18 MYH6 KIF3A MYO7A BCL2A1 CARD4 FAIM2 TNFRSF6B DCTN2 KIF11 MYH1 MYO3B IL10 MYH9 SPHK2 CASP5 BCL2L1 CARD6 HDAC1 DNAH1 KIF13A MYH10 MYO5A BAK1 MYO18B MYO1E CASP9 DNAH11 KIF13B MYH11 MYO5B BCL2L10 CARD9 HDAC3 MYO6 RIPK2 MALT1 BOK BCL2L13 CASP1 HRK DNAH12 KIF14 MYH13 MYO5C BCL2L2 CASP10 HTATIP2 DNAH14 KIF17 MYH13 MYO6 MYO3A BCL2L2 HDAC1 BAG3 BCL2L7P1 CASP10 IER3 DNAH14 KIF1B MYH2 MYO6 MYH3 BCL2L1 MYH11 CASP8 DNAH14 KIF1C MYH2 MYO7A BFAR CASP10 IER3 DFFB MYO15A MYH10 CASP2 DNAH14 KIF2 MYH3 MYO7B BID CASP2 IER3 DNAH14 KIF20A MYH4 MYO9A MYH7 SPP1 RTN4 ACTA1 BIK CASP2 IL10 DNAH17 KIF22 MYH4 MYO9B BIRC1 CASP2 IL18 FADD CFLAR DEDD ACTA2 DNAH2 KIF22 MYH6 OPA1 BIRC1 CASP4 IL2 ACTB DNAH5 MYO5A CARD11 DNAH3 KIF23 MYH6 OPA1 BIRC2 CASP5 MALT1 DNAH5 KIF23 MYH7 RPLP0P1 ACTC BCL2 BID FAIM2 BIRC3 CASP8 MCL1 DNAH7 KIF25 MYH7B RPLP0P1 BIRC6 BAX BIK APAF1 BIRC4 CASP9 NME6 DNAH8 KIF26A MYH8 TCTE1 IL2 BIRC5 CBX4 NOL3 DNAH9 KIF2C MYH9 TCTEL1 BIRC6 CFLAR PDCD2 DNAH9 KIF3A MYO10 TIAF1 FaGO

Annotaon List 1 List 2

GO:0006915 5 76 GO:0002684 6 14 GO:0048515 19 30

FaGO

Annotaon List 1 List 2

GO:0006915 5 76 GO:0002684 6 14 GO:0048515 19 30

Fisher exact test

Annotaon LOR p value FDR

GO:0006915 -4.13 8.41*10-30 3.6710-27 GO:0002684 -1.11 0.033 0.049 GO:0048515 -0.79 0.016 0.029

FaGO

Annotaon List 1 List 2

GO:0006915 5 76 GO:0002684 6 14 GO:0048515 19 30

Fisher exact test

Annotaon LOR p value FDR

GO:0006915 -4.13 8.41*10-30 3.6710-27 GO:0002684 -1.11 0.033 0.049 GO:0048515 -0.79 0.016 0.029

FaGO on Babelomics 5

Tool selecon Comparison

First list selecon

Second list selecon

Test opons Databases

Job informaon Data input

Gene list: bioformat“ID” Custom annotaon: bioformat Gene vs annotaon

1 Column: Gene ID Column 1: Column 2: Gene ID Funcon Reading results

Analysis overview

Results

Download results Reading results

Analysis overview

Results

Enriched class

Annotated genes from each list

Download results False Discovery Rate Reading results

Results visualizaon Babelomics 5: Gene Set Analysis Gene set enrichment

Comparison between a ranked list with user deﬁned annotaons Gene set enrichment

Comparison between a ranked list with user deﬁned annotaons

Ranked List: User provided Funconal annotaons: • Pre-deﬁned annotaons • Gene symbols • Custom annotaons • Probe Ids • Entrez Ids …

Mulple tesng context

Logisc regression model

Benjamini – Hochberg p-value correcon

Ranked results GSA: Logisc model

Annotaons 1 2 –

regression

Stasc Logisc

+ GSA: Logisc model

Annotaons 1 2 – P-value cutoﬀ 0.05

regression

Stasc Logisc

+ GSA: Logisc model

Annotaons 1 2 – P-value cutoﬀ 0.05

Block of genes enriched in annotaon 1

regression

Stasc Logisc

Block of genes enriched in annotaon 2

+ GSA on Babelomics 5

Tool selecon

Databases

Data selecon

Job informaon Data input

Gene list: bioformat“ranked ID” Custom annotaon: bioformat Gene vs annotaon

Column 1: Column 2: Column 2: Column 1: Ranked Gene ID Rank Funcon Gene ID Reading results

Analysis overview

Results

Download results Reading results

Analysis overview

Results

Enriched class

Annotated genes list

False Discovery Rate

Download results Reading results

Results visualizaon Keypoints

• FaGO: Detects signiﬁcant over representaon of funconal annotaons in one gene set respect to the other one. Keypoints

• FaGO: Detects signiﬁcant over representaon of funconal annotaons in one gene set respect to the other one. – Crical step: Gene list selecon. Tipically from diﬀerenal gene expression. • GSA: Detects gene sets (funconal annotaons) that are consistently associated to high or low values in a ranked list of genes. Training data and resources

Any quesons?

User: gda16 gda16b gda16c hp://babelomics.org Password: gda16 gda16b gda16c

FaGO hp://www.ncbi.nlm.nih.gov/pubmed/17478504 hp://www.ncbi.nlm.nih.gov/pubmed/14990455 wiki: hps://github.com/babelomics/babelomics/wiki/Single%20Enrichment Gene set analysis: Logisc model hp://www.ncbi.nlm.nih.gov/pubmed/20436964 hp://www.ncbi.nlm.nih.gov/pubmed/17407596 hp://www.ncbi.nlm.nih.gov/pubmed/15840702 wiki: hps://github.com/babelomics/babelomics/wiki/Gene%20Set%20Enrichment Thank you very much for your aenon!

Systems genomics team