methods_S5.Rmd

```{r}
#MISTAKE!! included normal patients in here as well. check PRAD as well
library(data.table);
library(dplyr);
library(RegParallel);
library(survminer)
```

```{r}
dataExpr = read.csv("01_voomExpr.csv");
rownames(dataExpr) = dataExpr$X;
dataExpr = dplyr::select(dataExpr,starts_with("TCGA"))
head(dataExpr[1:4]);
```


```{r}
genes_toKeep = read.csv("02_GSandMM.csv");
head(genes_toKeep[1:5]);

```
```{r}

```


```{r}
#genes_toKeep_list = genes_toKeep %>% filter(moduleColor == "yellow");
genes_toKeep_list = dplyr::filter(genes_toKeep,grepl("ITG",X))
ignore_integrins=c("ITGB3BP","ITGB1BP1")
genes_toKeep_list = genes_toKeep_list[!genes_toKeep_list$X %in% ignore_integrins,] #this line not in STAR protocols. remove itg
genes_toKeep_list = genes_toKeep_list$geneSymbol;

dataExpr = dataExpr[genes_toKeep_list,];
dataExpr
```

```{r}

```


```{r}
dataExpr = as.data.frame(t(dataExpr));
dataExpr = as.data.frame(scale(dataExpr));
head(dataExpr[1:4]);
```

```{r}
highExpr = 0.674;
lowExpr = -0.674;
dataExpr = as.data.frame(ifelse(dataExpr <= lowExpr,1,
                                ifelse(dataExpr >=highExpr,2,NA)));
dataExpr[] = lapply(dataExpr,factor);
dataExpr$SampleID = rownames(dataExpr)
```

```{r}
dataSurv = as.data.frame(fread("TCGA_survival_data"));
dataSurv$sample = gsub("-",".",dataSurv$sample);
dataSurv
```
```{r}
dataJoined = merge(dataSurv,dataExpr,by.x="sample",by.y = "SampleID");
```

```{r}
dataJoined
```


```{r}
temp4Hist = (dataJoined$OS.time)/365;

hist(temp4Hist,
     breaks = seq(from = 0, to = 24, by = 1),
     xaxt = 'n',
     xlab = "Years",
     main = "Histogram of OS.time");
axis(side = 1,
     at = seq(0,24,1),
     labels = seq(0,24,1));
```

```{r}
dataJoined = subset(dataJoined,OS.time <= 3650)
```

```{r}

```


```{r}
res = RegParallel(data = dataJoined,
                  formula = 'Surv(OS.time, OS) ~ [*]',
                  FUN = function(formula,data) coxph(formula = formula,
                                                     data = data,
                                                     ties = "breslow",
                                                     singular.ok = TRUE),
                  FUNtype = 'coxph',
                  variables = colnames(dataJoined)[10:ncol(dataJoined)],
                  blocksize = length(colnames(dataJoined)[10:ncol(dataJoined)]),
                  cores = 2,
                  nestedParallel = FALSE,
                  conflevel = 95);
res = res[!is.na(res$P), ];
res = res[order(res$LogRank, decreasing = FALSE)];
head(res);
```

```{r}
write.csv(res,"04_Survival.csv")
```

```{r}
dataDEG = read.csv("01_DEGsTreat.csv");
head(dataDEG)
```
```{r}
dataCytoscape = merge(res, dataDEG, by.x = "Variable",by.y = "X");
dataCytoscape = dplyr::select(dataCytoscape, Variable, LogRank,logFC,adj.P.Val);
head(dataCytoscape)
```
```{r}
write.csv(dataCytoscape, "04_CytoscapeInput.csv")
```

```{r}
res[order(res$LogRank, decreasing = FALSE),]
```


```{r}
shortList = subset(dataCytoscape,
                   LogRank < 0.05 & adj.P.Val <0.05);
shortList
```

```{r}
subset(res[grep("ITG",res$Variable),],LogRank<0.05 )
#integrins have significant effect on survival it seems
```


```{r}
subset(dataDEG[grep("ITG",dataDEG$X),][,c("X","adj.P.Val")],adj.P.Val<=0.05)
#however integrins are differentially expressed as i showed earlier
```

```{r}
geneSL = shortList$Variable;
for(i in 1:length(geneSL)){
  fit = survfit(as.formula(paste0("Surv(OS.time, OS) ~",geneSL[i])),
                data = dataJoined)
  print(
    ggsurvplot(fit,
               pval = TRUE,
               risk.table = TRUE,
               break.time.by = 365,
               ggtheme = theme_bw(),
               palette = c("blue","red"),
               xlim = c(0,3650),
               risk.table.y.text.col = TRUE,
               risk.table.y.text = FALSE,
               tables.theme = theme_cleantable(),
               tables.height = 0.15,
               xlab = "Days",
               font.x = c(12, "bold"),
               ylab = "Survival Probability",
               font.y = c(12,"bold"),
               legend.labs = c("Low-expression","High-expression"),
               font.legend = c(12,"bold"),
               legend.title = geneSL[i])
  )
}
```

```{r}
shortList_survonly = subset(dataCytoscape,
                   LogRank < 0.05);

```


```{r}
geneSL = shortList_survonly$Variable;
for(i in 1:length(geneSL)){
  fit = survfit(as.formula(paste0("Surv(OS.time, OS) ~",geneSL[i])),
                data = dataJoined)
  print(
    ggsurvplot(fit,
               pval = TRUE,
               risk.table = TRUE,
               break.time.by = 365,
               ggtheme = theme_bw(),
               palette = c("blue","red"),
               xlim = c(0,3650),
               risk.table.y.text.col = TRUE,
               risk.table.y.text = FALSE,
               tables.theme = theme_cleantable(),
               tables.height = 0.15,
               xlab = "Days",
               font.x = c(12, "bold"),
               ylab = "Survival Probability",
               font.y = c(12,"bold"),
               legend.labs = c("Low-expression","High-expression"),
               font.legend = c(12,"bold"),
               legend.title = geneSL[i])
  )
}
```


```{r}
122+119
```