1. Load TrendCatcher R package.

2. Now read the two demo master.list objects. since we want to compare the same biological pathway temporal behavior. We need to have two project TrendCatcher output: two master.list objects and one TimeHeatmap objects.

severe.path<-system.file("extdata", "MasterListSevere.rda", package = "TrendCatcher")
load(severe.path)
moderate.path<-system.file("extdata", "MasterListModerate.rda", package = "TrendCatcher")
load(moderate.path)
ht.path<-system.file("extdata", "htSevere.rda", package = "TrendCatcher")
load(ht.path)
#head(ht.severe$GO.df)
head(ht.severe$GO.df)
##           ID                              Description t.name    direction
## 1 GO:0006958 complement activation, classical pathway  0W-1W   Activation
## 2 GO:0006958 complement activation, classical pathway  1W-2W   Activation
## 3 GO:0006958 complement activation, classical pathway  2W-3W Deactivation
## 4 GO:0006958 complement activation, classical pathway  3W-4W Deactivation
## 5 GO:0006958 complement activation, classical pathway  4W-5W Deactivation
## 6 GO:0006958 complement activation, classical pathway  5W-6W Deactivation
##   Avg_log2FC n_total n_background n_up n_down
## 1  2.1002689      77          141   77      0
## 2  0.6301674      32          141   32      0
## 3 -0.2587066      52          141   20     32
## 4 -0.3352649      62          141   16     46
## 5 -0.1453267      67          141   15     52
## 6 -0.5720138      68          141    0     68
##                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            geneID_up
## 1 CLU/SERPING1/C1QB/CD55/CR1/IGKC/IGKV4-1/IGKV3D-20/IGKV3D-11/IGLV6-57/IGLV1-51/IGLV1-47/IGLV1-44/IGLV7-43/IGLV1-40/IGLV3-27/IGLV3-25/IGLV2-23/IGLV3-21/IGLV3-19/IGLV2-14/IGLV2-11/IGLV3-1/IGLC2/IGLC3/IGLC7/IGHA2/IGHG4/IGHG2/IGHA1/IGHG1/IGHG3/IGHV6-1/IGHV1-3/IGHV2-5/IGHV3-7/IGHV3-11/IGHV3-15/IGHV1-18/IGHV3-20/IGHV3-21/IGHV3-23/IGHV1-24/IGHV3-33/IGHV4-34/IGHV4-39/IGHV3-48/IGHV5-51/IGHV3-53/IGHV3-66/IGHV1-69/IGHV2-70D/IGLC6/IGHV4-59/IGHV3-74/IGHV3-72/IGHV4-31/IGKV2D-30/IGKV3-20/IGKV1D-33/IGKV1-17/IGKV1-16/IGKV1-39/IGKV2D-28/IGKV2-30/IGKV1-12/IGKV1-5/IGKV2-28/IGKV3-15/IGKV1D-39/IGHV3-30/IGHV2-70/IGHV4-4/IGLV2-8/IGKV1D-12/IGHV3-64D/IGHV5-10-1
## 2                                                                                                                                                                                                                                                                                                                                                                                        CLU/SERPING1/CD55/CR1/IGKC/IGKV4-1/IGKV3D-20/IGLV3-27/IGLV3-25/IGLV2-14/IGLV2-11/IGLV3-1/IGHA2/IGHV6-1/IGHV3-7/IGHV3-15/IGHV1-18/IGHV3-23/IGHV1-24/IGHV3-48/IGHV2-70D/IGHV3-74/IGHV3-72/IGKV2D-30/IGKV3-20/IGKV2D-28/IGKV2-30/IGKV1-5/IGKV2-28/IGHV2-70/IGHV1-69D/IGHV3-64D
## 3                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               CLU/CD55/IGKC/IGKV4-1/IGKV3D-20/IGLV2-11/IGHA2/IGHV6-1/IGHV3-7/IGHV3-15/IGHV1-18/IGHV3-48/IGHV2-70D/IGHV3-74/IGHV3-72/IGKV2D-30/IGKV2D-28/IGKV2-30/IGKV1-5/IGHV3-64D
## 4                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                          IGKV4-1/IGKV3D-20/TRBC2/IGHA2/IGHV6-1/IGHV3-7/IGHV3-15/IGHV1-18/IGHV3-48/IGHV3-74/IGHV3-72/IGKV2D-30/IGKV2D-28/IGKV2-30/IGKV1-5/IGHV3-64D
## 5                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                       IGKV4-1/IGLV2-14/TRBC1/TRBC2/IGHA2/IGHV3-7/IGHV3-15/IGHV3-48/IGHV3-74/IGHV3-72/IGKV3-20/IGKV2D-28/IGKV1-5/IGKV2-28/IGHV3-64D
## 6                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
##                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           geneID_down
## 1                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
## 2                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                    
## 3                                                                                                                                                                                                                                                                                                                   SERPING1/C1QB/IGLV6-57/IGLV1-47/IGLV1-44/IGLV1-40/IGLV3-27/IGLV3-25/IGLV2-23/IGLV3-21/IGLV3-19/IGLV2-14/IGLC2/IGLC3/IGHG4/IGHG1/IGHG3/IGHV3-20/IGHV3-33/IGHV4-34/IGHV4-39/IGHV3-53/IGHV1-69/IGHV4-59/IGKV3-20/IGKV1D-33/IGKV1-16/IGKV1-39/IGKV1D-39/IGHV3-30/IGHV1-69D/IGHV5-10-1
## 4                                                                                                                                                                                                  CLU/SERPING1/C1QB/CR1/IGLV6-57/IGLV1-47/IGLV1-44/IGLV1-40/IGLV3-27/IGLV3-25/IGLV2-23/IGLV3-21/IGLV3-19/IGLV3-1/IGLC2/IGLC3/IGHG4/IGHG1/IGHG3/IGHV1-3/IGHV2-5/IGHV3-20/IGHV3-21/IGHV1-24/IGHV3-33/IGHV4-34/IGHV4-39/IGHV5-51/IGHV3-53/IGHV1-69/IGHV2-70D/IGHV4-59/IGHV4-31/IGKV3-20/IGKV1D-33/IGKV1-16/IGKV1-39/IGKV1-12/IGKV1D-39/IGHV3-30/IGHV2-70/IGHV4-4/IGLV2-8/IGKV1D-12/IGHV1-69D/IGHV5-10-1
## 5                                                                                                                                                 CLU/SERPING1/C1QB/CR1/IGKV3D-11/IGLV6-57/IGLV1-47/IGLV1-44/IGLV1-40/IGLV3-27/IGLV3-25/IGLV2-23/IGLV3-21/IGLV3-19/IGLV3-1/IGLC2/IGLC3/IGHG4/IGHA1/IGHG1/IGHG3/IGHV1-3/IGHV2-5/IGHV3-11/IGHV3-20/IGHV3-21/IGHV1-24/IGHV3-33/IGHV4-34/IGHV4-39/IGHV5-51/IGHV3-53/IGHV3-66/IGHV1-69/IGHV2-70D/IGLC6/IGHV4-59/IGHV4-31/IGKV1D-33/IGKV1-17/IGKV1-16/IGKV1-39/IGKV1-12/IGKV3-15/IGKV1D-39/IGHV3-30/IGHV2-70/IGHV4-4/IGLV2-8/IGKV1D-12/IGHV1-69D/IGHV5-10-1
## 6 SERPING1/C1QB/CR1/IGKC/IGKV3D-20/IGKV3D-11/IGLV6-57/IGLV1-51/IGLV1-47/IGLV1-44/IGLV7-43/IGLV1-40/IGLV3-27/IGLV3-25/IGLV2-23/IGLV3-21/IGLV3-19/IGLV2-14/IGLV2-11/IGLV3-1/IGLC2/IGLC3/IGLC7/IGHG4/IGHA1/IGHG1/IGHG3/IGHV1-3/IGHV2-5/IGHV3-11/IGHV1-18/IGHV3-20/IGHV3-21/IGHV3-23/IGHV1-24/IGHV3-33/IGHV4-34/IGHV4-39/IGHV3-48/IGHV5-51/IGHV3-53/IGHV3-66/IGHV1-69/IGHV2-70D/IGLC6/IGHV4-59/IGHV3-74/IGHV4-31/IGKV2D-30/IGKV3-20/IGKV1D-33/IGKV1-17/IGKV1-16/IGKV1-39/IGKV2D-28/IGKV2-30/IGKV1-12/IGKV1-5/IGKV2-28/IGKV3-15/IGKV1D-39/IGHV3-30/IGHV2-70/IGHV4-4/IGLV2-8/IGKV1D-12/IGHV1-69D/IGHV5-10-1
##            p.adjust.up        p.adjust.down
## 1 6.67222905253988e-72                     
## 2 2.19506922004502e-18                     
## 3 1.63402445206667e-08  9.9477266204752e-33
## 4 2.74522759489778e-05 7.55610755399035e-46
## 5 2.69045301915842e-05  3.3223517232817e-48
## 6                       6.7007249596023e-69

3. To compare DDEGs trajectories from a specific enriched biological pathway from two groups. In this example, we show DDEGs from “neutrophil activation”, which is one of the top enriched timeheatmap biological pathway from severe group. We want to see these DDEGs trajector comparison from severe group and moderate group, since both groups record the temporal behavior of these genes.

ht.severe$merge.df %>% filter(Description == "neutrophil activation")
##                          ID           Description     p.adjust GeneRatio
## 0W-1W.GO:0042119 GO:0042119 neutrophil activation 3.063434e-07    12/644
## 1W-2W.GO:0042119 GO:0042119 neutrophil activation 4.321727e-07    11/503
## 3W-4W.GO:0042119 GO:0042119 neutrophil activation 5.329931e-03     5/282
## 4W-5W.GO:0042119 GO:0042119 neutrophil activation 4.305079e-05     8/386
## 5W-6W.GO:0042119 GO:0042119 neutrophil activation 1.237944e-07    11/458
## 6W-7W.GO:0042119 GO:0042119 neutrophil activation 1.786931e-04     8/452
##                   BgRatio
## 0W-1W.GO:0042119 41/18800
## 1W-2W.GO:0042119 41/18800
## 3W-4W.GO:0042119 41/18800
## 4W-5W.GO:0042119 41/18800
## 5W-6W.GO:0042119 41/18800
## 6W-7W.GO:0042119 41/18800
##                                                                                geneID
## 0W-1W.GO:0042119 TYROBP/GRN/SPI1/CTSG/PRAM1/ANXA3/FCER1G/PRKCD/CAMP/ITGAM/MYD88/CD177
## 1W-2W.GO:0042119       TYROBP/GRN/SPI1/CTSG/ANXA1/ANXA3/PRKCD/CAMP/CD300A/ITGAM/MYD88
## 3W-4W.GO:0042119                                          GRN/SPI1/CD300A/MYD88/CD177
## 4W-5W.GO:0042119                      SPI1/CTSG/FCER1G/PRKCD/CD300A/ITGAM/MYD88/CD177
## 5W-6W.GO:0042119     SPI1/CTSG/PRAM1/ANXA3/FCER1G/PRKCD/CAMP/CD300A/ITGAM/MYD88/CD177
## 6W-7W.GO:0042119                        SPI1/CTSG/PRAM1/ANXA3/CAMP/CD300A/ITGAM/CD177
##                  t.name         type
## 0W-1W.GO:0042119  0W-1W   Activation
## 1W-2W.GO:0042119  1W-2W   Activation
## 3W-4W.GO:0042119  3W-4W Deactivation
## 4W-5W.GO:0042119  4W-5W Deactivation
## 5W-6W.GO:0042119  5W-6W Deactivation
## 6W-7W.GO:0042119  6W-7W Deactivation

To show the DDEGs from severe group which were enriched for neutrophil activation pathway, and how their temporal behavior differs between moderate and severe group. We applied LOESS smoothing curve fitting method.

g<-draw_CurveComp(master.list.1 = master.list.severe, master.list.2 = master.list.moderate, ht.1 = ht.severe, pathway = "neutrophil activation",group.1.name = "severe", group.2.name = "moderate")
## Found 117 DDEGs from neutrophil activation

To quantify which time interval these two group separate, we applied permutation test and marked the significant separation time in grey.

perm_output<-draw_CurveComp_Perm(master.list.1 = master.list.severe, 
                                 master.list.2 = master.list.moderate, 
                                 ht.1 = ht.severe, 
                                 pathway = "neutrophil activation", 
                                 group.1.name = "severe", 
                                 group.2.name = "moderate", 
                                 n.perm = 100, 
                                 parall = FALSE, 
                                 pvalue.threshold = 0.05)
names(perm_output)
## [1] "adjusted.pvalue.area" "perm"                 "st"                  
## [4] "en"                   "plot"
perm_output$plot