diff --git a/01_advanced.R b/01_advanced.R new file mode 100644 index 0000000..839ee81 --- /dev/null +++ b/01_advanced.R @@ -0,0 +1,66 @@ +library(shiny) +library(dplyr) +library(tidyr) +library(ggplot2) + +expr_table <- read.table("expression_matrix_w_metadata.txt", sep = ",", header = TRUE, stringsAsFactors = FALSE) +metacols <- grep("^ENSG", colnames(expr_table), invert = TRUE, value = TRUE) + +allgenes <- read.table("mart_export.txt", sep = "\t", header = TRUE, stringsAsFactors = FALSE) +allgenes <- allgenes[allgenes$ensId %in% colnames(expr_table),] +lookuptable <- unstack(allgenes) + +ui <- fluidPage( + + titlePanel("Immune Cell Gene Expression Data"), + + sidebarLayout( + sidebarPanel( + selectizeInput("ingene", "Choose a gene:", choices = sort(allgenes$symbol), selected = "HBB"), + helpText(h4("Data source:")), + helpText("Linsley PS, Speake C, Whalen E, Chaussabel D.", em("Copy number loss of the interferon gene cluster in melanomas is linked to reduced T cell infiltrate and poor patient prognosis."), + "PLoS One 2014 Oct 14;9(10):e109760."), + a(href = "https://www.ncbi.nlm.nih.gov/pubmed/25314013", "On PubMed"), br(), + a(href = "https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE60424", "On GEO") + ), + mainPanel( + plotOutput("theplot"), + helpText("Expression values were normalized using the TMM (trimmed mean of m-value) method.", + "This means that counts have been normalized by library size but not gene length.", + "Plot shows Tukey boxplots: whiskers extend to the highest/lowest value within the ", + "box limit +/- 1.5 * IQR. Samples beyond that range are plotted as points.") + ) + ) +) + +server <- function(input, output) { + + output$theplot <- renderPlot({ + req(input$ingene) + + ens <- lookuptable[[input$ingene]] + gene <- input$ingene + + tinyframe <- expr_table[,c(ens, metacols)] + tinyframe <- tinyframe %>% gather(variable, value, one_of(ens)) + p <- ggplot(tinyframe, aes(x = variable, y = value, color = cellType)) + + ggtitle(paste0(gene, " expression by cell type across conditions")) + + geom_boxplot() + + ylab("Normalized Counts") + + theme(axis.text.x = element_blank(), + axis.ticks.x = element_blank(), + axis.title.x = element_blank()) + + if (length(ens) > 1) { + p <- p + facet_grid(variable ~ diseaseStatus, scales = "free_y") + } else { + p <- p + facet_grid(~ diseaseStatus) + } + return(p) + }) + +} + +# Run the application +shinyApp(ui = ui, server = server) + diff --git a/01_simple.R b/01_simple.R new file mode 100644 index 0000000..396046b --- /dev/null +++ b/01_simple.R @@ -0,0 +1,25 @@ +library(shiny) + + +ui <- fluidPage( + + fluidRow( + column(width = 12, + textInput("textinputlabel", "What's up?"), + textOutput("newtext") + + ) + ) +) + +server <- function(input, output) { + + output$newtext <- renderText({ + toupper(input$textinputlabel) + }) + +} + +# Run the application +shinyApp(ui = ui, server = server) + diff --git a/02_advanced.R b/02_advanced.R new file mode 100644 index 0000000..3826d17 --- /dev/null +++ b/02_advanced.R @@ -0,0 +1,5 @@ +library(profvis) + +# Run the application +profvis(runApp("01_advanced.R")) + diff --git a/02_simple.R b/02_simple.R new file mode 100644 index 0000000..23d5ff4 --- /dev/null +++ b/02_simple.R @@ -0,0 +1,45 @@ +library(shiny) +library(ggplot2) +library(dplyr) +library(tidyr) + +ui <- fluidPage( + + fluidRow( + column(width = 12, + sliderInput("sample_count", "How many times should we sample each population?", + value = 10, + min = 1, + max = 500), + plotOutput("sample_plot"), + tableOutput("sample_table") + ) + ) +) + +server <- function(input, output) { + + sample_dat <- reactive({ + data.frame("rowid" = paste0("row", seq(1,input$sample_count)), + "thicc bois" = rnorm(input$sample_count, 0, 1), + "chonkers" = rnorm(input$sample_count, 0.2, 1), + stringsAsFactors = FALSE, + check.names = FALSE) %>% + gather(population, measurement, -rowid) + }) + + output$sample_plot <- renderPlot({ + ggplot(sample_dat(), aes(x = population, y = measurement, fill = population)) + + geom_boxplot() + + geom_point(position = position_jitterdodge(0.5)) + + theme(axis.text = element_text(size = rel(2)), + axis.title = element_text(size = rel(2)), + legend.text = element_text(size = rel(1.5)), + legend.title = element_text(size = rel(1.5))) + }) + +} + +# Run the application +shinyApp(ui = ui, server = server) + diff --git a/03_advanced.R b/03_advanced.R new file mode 100644 index 0000000..490d481 --- /dev/null +++ b/03_advanced.R @@ -0,0 +1,100 @@ +library(shiny) +library(dplyr) +library(tidyr) +library(ggplot2) +library(RSQLite) + +conn <- dbConnect(RSQLite::SQLite(), "GSE60424.db") +choosenames <- dbGetQuery(conn, "SELECT symbol + FROM identifiers i + INNER JOIN expr e + ON i.ensId = e.genenames;") +choosenames <- choosenames[[1]] +dbDisconnect(conn) + +ui <- fluidPage( + + titlePanel("Immune Cell Gene Expression Data"), + + sidebarLayout( + sidebarPanel( + selectizeInput("ingene", "Choose a gene:", choices = NULL, selected = NULL), + helpText(h4("Data source:")), + helpText("Linsley PS, Speake C, Whalen E, Chaussabel D.", em("Copy number loss of the interferon gene cluster in melanomas is linked to reduced T cell infiltrate and poor patient prognosis."), + "PLoS One 2014 Oct 14;9(10):e109760."), + a(href = "https://www.ncbi.nlm.nih.gov/pubmed/25314013", "On PubMed"), br(), + a(href = "https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE60424", "On GEO") + ), + mainPanel( + plotOutput("theplot"), + helpText("Expression values were normalized using the TMM (trimmed mean of m-value) method.", + "This means that counts have been normalized by library size but not gene length.", + "Plot shows Tukey boxplots: whiskers extend to the highest/lowest value within the ", + "box limit +/- 1.5 * IQR. Samples beyond that range are plotted as points.") + ) + ) +) + +server <- function(input, output, session) { + + updateSelectizeInput(session, "ingene", choices = choosenames, selected = "HBB", server = TRUE) + + makeTinyframe <- function(gene, dbpath){ + conn <- dbConnect(RSQLite::SQLite(), dbpath) + meta <- dbGetQuery(conn, "SELECT * FROM meta;") + exprquery <- sprintf("SELECT i.symbol, e.* + FROM identifiers i + JOIN expr e + ON i.ensId = e.genenames + WHERE i.symbol == %s;", shQuote(gene)) + expr <- dbGetQuery(conn, exprquery) + dbDisconnect(conn) + + rownames(expr) <- expr$genenames + togene <- expr[,grep("lib", colnames(expr), invert = TRUE)] + expr <- expr[,grep("lib", colnames(expr))] + expr <- t(expr) + + tinyframe <- merge(meta, expr, by.x = "library", by.y = 0) + tinyframe <- tinyframe %>% + gather(gene, TPM, starts_with("ENS")) %>% + merge(., togene, by.x = "gene", by.y = "genenames") %>% + rename(genename = symbol) + return(tinyframe) + + } + + makePlot <- function(tinyframe){ + + printgene <- unique(tinyframe$genename) + + p <- ggplot(tinyframe, aes(x = genename, y = TPM, color = celltype)) + + ggtitle(paste0(printgene, " expression by cell type across conditions")) + + geom_boxplot() + + theme(axis.text.x = element_blank(), axis.ticks.x = element_blank(), axis.title.x = element_blank()) + + if (length(unique(tinyframe$gene)) > 1) { + p <- p + facet_grid(variable ~ disease_status, scales = "free_y") + } else { + p <- p + facet_grid(~ disease_status) + } + return(p) + } + + inputgene <- reactive({ + validate( + need(input$ingene != "", "Please select a gene") + ) + input$ingene + }) + + output$theplot <- renderPlot({ + tf <- makeTinyframe(inputgene(), "GSE60424.db") + makePlot(tf) + }) + +} + +# Run the application +shinyApp(ui = ui, server = server) + diff --git a/03_simple.R b/03_simple.R new file mode 100644 index 0000000..084a9d3 --- /dev/null +++ b/03_simple.R @@ -0,0 +1,57 @@ +library(shiny) +library(ggplot2) +library(dplyr) +library(tidyr) + +ui <- fluidPage( + + fluidRow( + column(width = 12, + sliderInput("sample_count", "How many times should we sample each population?", + value = 10, + min = 1, + max = 500), + plotOutput("sample_plot"), + textOutput("sample_result") + ) + ) +) + +server <- function(input, output) { + + sample_dat <- reactive({ + data.frame("rowid" = paste0("row", seq(1,input$sample_count)), + "thicc bois" = rnorm(input$sample_count, 0, 1), + "chonkers" = rnorm(input$sample_count, 0.2, 1), + stringsAsFactors = FALSE, + check.names = FALSE) + }) + + output$sample_plot <- renderPlot({ + sample_dat() %>% + gather(population, measurement, -rowid) %>% + ggplot(aes(x = population, y = measurement, fill = population)) + + geom_boxplot() + + geom_point(position = position_jitterdodge(0.5)) + + theme(axis.text = element_text(size = rel(2)), + axis.title = element_text(size = rel(2)), + legend.text = element_text(size = rel(1.5)), + legend.title = element_text(size = rel(1.5))) + }) + + output$sample_result <- renderText({ + p <- t.test(sample_dat()[["thicc bois"]], + sample_dat()[["chonkers"]], + paired = FALSE)[["p.value"]] + if(p >= 0.05){ + paste("p-value = ", p) + } else { + paste("p-value = ", p, "*") + } + }) + +} + +# Run the application +shinyApp(ui = ui, server = server) + diff --git a/04_simple.R b/04_simple.R new file mode 100644 index 0000000..d4bf8b6 --- /dev/null +++ b/04_simple.R @@ -0,0 +1,64 @@ +library(shiny) +library(ggplot2) +library(dplyr) +library(tidyr) + +ui <- fluidPage( + + fluidRow( + column(width = 12, + actionButton("do_sample", "Moar!"), + plotOutput("sample_plot"), + textOutput("sample_result") + ) + ) +) + +server <- function(input, output) { + + sample_count <- reactiveVal(10) + + observeEvent(input$do_sample,{ + x <- sample_count() + 10 + sample_count(x) + }) + + sample_dat <- reactive({ + tmp <- sample_count() + data.frame("rowid" = paste0("row", seq(1, tmp)), + "thicc bois" = rnorm(tmp, 0, 1), + "chonkers" = rnorm(tmp, 0.2, 1), + stringsAsFactors = FALSE, + check.names = FALSE) + }) + + output$sample_plot <- renderPlot({ + sample_dat() %>% + gather(population, measurement, -rowid) %>% + ggplot(aes(x = population, y = measurement, fill = population)) + + geom_boxplot() + + geom_point(position = position_jitterdodge(0.5)) + + ggtitle(paste0("You sampled ", nrow(sample_dat()), " absolute units")) + + theme(axis.text = element_text(size = rel(2)), + axis.title = element_text(size = rel(2)), + legend.text = element_text(size = rel(1.5)), + legend.title = element_text(size = rel(1.5)), + plot.title = element_text(size = rel(2))) + }) + + output$sample_result <- renderText({ + p <- t.test(sample_dat()[["thicc bois"]], + sample_dat()[["chonkers"]], + paired = FALSE)[["p.value"]] + if(p >= 0.05){ + paste("p-value = ", p) + } else { + paste("p-value = ", p, "*") + } + }) + +} + +# Run the application +shinyApp(ui = ui, server = server) + diff --git a/GSE60424.db b/GSE60424.db new file mode 100644 index 0000000..346aabe Binary files /dev/null and b/GSE60424.db differ diff --git a/expression_matrix_w_metadata.txt b/expression_matrix_w_metadata.txt new file mode 100644 index 0000000..de6c354 --- /dev/null +++ b/expression_matrix_w_metadata.txt @@ -0,0 +1,135 @@ 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Blood","Healthy Control",44,"F","Hispanic" 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Blood","MS pretreatment",31,"F","White" 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Blood","MS posttreatment",33,"F","White" 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Blood","Type 1 Diabetes",34,"F","White" 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Blood","Type 1 Diabetes",37,"F","White" 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posttreatment",33,"F","White" 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posttreatment",33,"F","White" 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1 Diabetes",34,"F","White" 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Blood","Sepsis",43,"M","White" 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Blood","MS pretreatment",45,"F","White" 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posttreatment",46,"F","White" 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posttreatment",46,"F","White" 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posttreatment",46,"F","White" 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Blood","Healthy Control",20,"F","White" 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Blood","Healthy Control",21,"F","White" 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Blood","Healthy Control",53,"F","White" 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Blood","Type 1 Diabetes",40,"M","White" 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1 Diabetes",41,"F","White" 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Blood","Type 1 Diabetes",41,"F","White" 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Blood","MS pretreatment",55,"F","White" 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posttreatment",56,"F","White" 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posttreatment",56,"F","White" 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posttreatment",56,"F","White" 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posttreatment",56,"F","White" 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Blood","ALS",58,"--","--" diff --git a/mart_export.txt b/mart_export.txt new file mode 100644 index 0000000..65cfa5e --- /dev/null +++ b/mart_export.txt @@ -0,0 +1,63306 @@ +ensId symbol +ENSG00000252303 RNU6-280P +ENSG00000281771 Y_RNA +ENSG00000281256 RP11-222G7.2 +ENSG00000283272 Clostridiales-1 +ENSG00000280864 RP11-654C22.2 +ENSG00000280792 RP11-315F22.1 +ENSG00000282878 RP11-399E6.1 +ENSG00000283276 ABBA01000934.1 +ENSG00000281822 RNU1-62P +ENSG00000281384 AC093802.1 +ENSG00000280505 RP11-654C22.1 +ENSG00000281764 RP11-399E6.2 +ENSG00000281316 DPPA2P2 +ENSG00000280963 SERTAD4-AS1 +ENSG00000280775 RNA5SP136 +ENSG00000281876 RP11-399E6.4 +ENSG00000281766 RYBP +ENSG00000281518 FOXO6 +ENSG00000281614 INPP5D +ENSG00000280584 OBP2B +ENSG00000281230 SERTAD4 +ENSG00000281917 SLC16A1 +ENSG00000281067 ST13P19 +ENSG00000281552 RP4-667H12.4 +ENSG00000280820 LCN1P1 +ENSG00000280751 AKR7A2P1 +ENSG00000280632 Y_RNA +ENSG00000281557 RP11-31F15.2 +ENSG00000281430 LCN1P2 +ENSG00000281813 KAT6B 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RP11-675M1.2 +ENSG00000282928 MAP4K1 +ENSG00000282050 IGKV7-3 +ENSG00000282699 PGBD4P5 +ENSG00000282974 DPP6 +ENSG00000282448 IGKV2-4 +ENSG00000282801 IGKV1-5 +ENSG00000282163 IGKV1-6 +ENSG00000282310 IGKV3-7 +ENSG00000283113 IGKV1-8 +ENSG00000282758 IGKV1-9 +ENSG00000282248 IGKV2-10 +ENSG00000282823 IGKV3-11 +ENSG00000282694 IGKV1-12 +ENSG00000282603 IGKV1-13 +ENSG00000282251 IGKV2-14 +ENSG00000282447 IGKV3-15 +ENSG00000282282 IGKV1-16 +ENSG00000281978 IGKV1-17 +ENSG00000282200 IGKV2-18 +ENSG00000282244 IGKV2-19 +ENSG00000282402 IGKV3-20 +ENSG00000282210 IGKV6-21 +ENSG00000282762 IGKV1-22 +ENSG00000282093 IGKV2-23 +ENSG00000282671 IGKV2-24 +ENSG00000282062 IGKV3-25 +ENSG00000282622 IGKV2-26 +ENSG00000282000 RP11-421K23.1 +ENSG00000282666 IGKV1-27 +ENSG00000283068 IBTK +ENSG00000282025 IGKV2-28 +ENSG00000282635 IGKV2-29 +ENSG00000282956 MAGI1 +ENSG00000281933 IGKV2-30 +ENSG00000282743 IGKV3-31 +ENSG00000282571 IGKV1-32 +ENSG00000282811 IGKV1-33 +ENSG00000282149 IGKV3-34 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