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29.5 kB
| # Calling Cards Region Quantification | |
| # Quantifies enrichment of insertions/hops in genomic regions | |
| # | |
| # This script: | |
| # 1. Counts insertions overlapping each genomic region (experiment and background) | |
| # 2. Calculates enrichment scores | |
| # 3. Computes Poisson and hypergeometric p-values | |
| # | |
| # Works with any data in BED3+ format (chr, start, end, ...) | |
| # For calling cards: each insertion is counted once regardless of depth | |
| # | |
| # COORDINATE SYSTEMS: | |
| # - Input BED files are assumed to be 0-indexed, half-open [start, end) | |
| # - GenomicRanges uses 1-indexed, closed [start, end] | |
| # - Conversion: GR_start = BED_start + 1, GR_end = BED_end | |
| # | |
| # Usage: | |
| # Rscript quantify_regions.R | |
| library(tidyverse) | |
| library(GenomicRanges) | |
| # Statistical Functions --------------------------------------------------- | |
| #' Calculate enrichment (calling cards effect) | |
| #' | |
| #' @param total_background_hops Total number of hops in background (scalar or vector) | |
| #' @param total_experiment_hops Total number of hops in experiment (scalar or vector) | |
| #' @param background_hops Number of hops in background per region (vector) | |
| #' @param experiment_hops Number of hops in experiment per region (vector) | |
| #' @param pseudocount Pseudocount to avoid division by zero (default: 0.1) | |
| #' @return Enrichment values | |
| calculate_enrichment <- function(total_background_hops, | |
| total_experiment_hops, | |
| background_hops, | |
| experiment_hops, | |
| pseudocount = 0.1) { | |
| # Input validation | |
| if (!all(is.numeric(c( | |
| total_background_hops, total_experiment_hops, | |
| background_hops, experiment_hops | |
| )))) { | |
| stop("All inputs must be numeric") | |
| } | |
| # Get the length of the region vectors | |
| n_regions <- length(background_hops) | |
| # Ensure experiment_hops is same length as background_hops | |
| if (length(experiment_hops) != n_regions) { | |
| stop("background_hops and experiment_hops must be the same length") | |
| } | |
| # Recycle scalar totals to match region length if needed | |
| if (length(total_background_hops) == 1) { | |
| total_background_hops <- rep(total_background_hops, n_regions) | |
| } | |
| if (length(total_experiment_hops) == 1) { | |
| total_experiment_hops <- rep(total_experiment_hops, n_regions) | |
| } | |
| # Now check all are same length | |
| if (length(total_background_hops) != n_regions || | |
| length(total_experiment_hops) != n_regions) { | |
| stop("All input vectors must be the same length or scalars") | |
| } | |
| # Calculate enrichment | |
| numerator <- experiment_hops / total_experiment_hops | |
| denominator <- (background_hops + pseudocount) / total_background_hops | |
| enrichment <- numerator / denominator | |
| # Check for invalid values | |
| if (any(enrichment < 0, na.rm = TRUE)) { | |
| stop("Enrichment values must be non-negative") | |
| } | |
| if (any(is.na(enrichment))) { | |
| stop("Enrichment values must not be NA") | |
| } | |
| if (any(is.infinite(enrichment))) { | |
| stop("Enrichment values must not be infinite") | |
| } | |
| return(enrichment) | |
| } | |
| #' Calculate Poisson p-values | |
| #' | |
| #' @param total_background_hops Total number of hops in background (scalar or vector) | |
| #' @param total_experiment_hops Total number of hops in experiment (scalar or vector) | |
| #' @param background_hops Number of hops in background per region (vector) | |
| #' @param experiment_hops Number of hops in experiment per region (vector) | |
| #' @param pseudocount Pseudocount for lambda calculation (default: 0.1) | |
| #' @param ... additional arguments to `ppois`. note that lower tail is set to FALSE | |
| #' already | |
| #' @return Poisson p-values | |
| calculate_poisson_pval <- function(total_background_hops, | |
| total_experiment_hops, | |
| background_hops, | |
| experiment_hops, | |
| pseudocount = 0.1, | |
| ...) { | |
| # Input validation | |
| if (!all(is.numeric(c( | |
| total_background_hops, total_experiment_hops, | |
| background_hops, experiment_hops | |
| )))) { | |
| stop("All inputs must be numeric") | |
| } | |
| # Get the length of the region vectors | |
| n_regions <- length(background_hops) | |
| # Ensure experiment_hops is same length as background_hops | |
| if (length(experiment_hops) != n_regions) { | |
| stop("background_hops and experiment_hops must be the same length") | |
| } | |
| # Recycle scalar totals to match region length if needed | |
| if (length(total_background_hops) == 1) { | |
| total_background_hops <- rep(total_background_hops, n_regions) | |
| } | |
| if (length(total_experiment_hops) == 1) { | |
| total_experiment_hops <- rep(total_experiment_hops, n_regions) | |
| } | |
| # Now check all are same length | |
| if (length(total_background_hops) != n_regions || | |
| length(total_experiment_hops) != n_regions) { | |
| stop("All input vectors must be the same length or scalars") | |
| } | |
| # Calculate hop ratio | |
| hop_ratio <- total_experiment_hops / total_background_hops | |
| # Calculate expected number of hops (mu/lambda parameter) | |
| # Add pseudocount to avoid mu = 0 | |
| mu <- (background_hops + pseudocount) * hop_ratio | |
| # Observed hops in experiment | |
| x <- experiment_hops | |
| # Calculate p-value: P(X >= x) = 1 - P(X < x) = 1 - P(X <= x-1) | |
| # This is equivalent to: 1 - CDF(x) + PMF(x) | |
| # Using the upper tail directly with lower.tail = FALSE | |
| pval <- ppois(x - 1, lambda = mu, lower.tail = FALSE, ...) | |
| return(pval) | |
| } | |
| #' Calculate hypergeometric p-values | |
| #' | |
| #' @param total_background_hops Total number of hops in background (scalar or vector) | |
| #' @param total_experiment_hops Total number of hops in experiment (scalar or vector) | |
| #' @param background_hops Number of hops in background per region (vector) | |
| #' @param experiment_hops Number of hops in experiment per region (vector) | |
| #' @param ... additional arguments to phyper. Note that lower tail is set to | |
| #' false already | |
| #' @return Hypergeometric p-values | |
| calculate_hypergeom_pval <- function(total_background_hops, | |
| total_experiment_hops, | |
| background_hops, | |
| experiment_hops, | |
| ...) { | |
| # Input validation | |
| if (!all(is.numeric(c( | |
| total_background_hops, total_experiment_hops, | |
| background_hops, experiment_hops | |
| )))) { | |
| stop("All inputs must be numeric") | |
| } | |
| # Get the length of the region vectors | |
| n_regions <- length(background_hops) | |
| # Ensure experiment_hops is same length as background_hops | |
| if (length(experiment_hops) != n_regions) { | |
| stop("background_hops and experiment_hops must be the same length") | |
| } | |
| # Recycle scalar totals to match region length if needed | |
| if (length(total_background_hops) == 1) { | |
| total_background_hops <- rep(total_background_hops, n_regions) | |
| } | |
| if (length(total_experiment_hops) == 1) { | |
| total_experiment_hops <- rep(total_experiment_hops, n_regions) | |
| } | |
| # Now check all are same length | |
| if (length(total_background_hops) != n_regions || | |
| length(total_experiment_hops) != n_regions) { | |
| stop("All input vectors must be the same length or scalars") | |
| } | |
| # Hypergeometric parameters | |
| # M: total number of balls (total hops) | |
| M <- total_background_hops + total_experiment_hops | |
| # n: number of white balls (experiment hops) | |
| n <- total_experiment_hops | |
| # N: number of draws (hops in region) | |
| N <- background_hops + experiment_hops | |
| # x: number of white balls drawn (experiment hops in region) - 1 for upper tail | |
| x <- experiment_hops - 1 | |
| # Handle edge cases | |
| valid <- (M >= 1) & (N >= 1) | |
| pval <- rep(1, length(M)) | |
| # Calculate p-value for valid cases: P(X >= x) = 1 - P(X <= x-1) | |
| if (any(valid)) { | |
| pval[valid] <- phyper(x[valid], n[valid], M[valid] - n[valid], N[valid], | |
| lower.tail = FALSE, ... | |
| ) | |
| } | |
| return(pval) | |
| } | |
| # GRanges Conversion Functions -------------------------------------------- | |
| #' Convert BED format data frame to GRanges | |
| #' | |
| #' Handles coordinate system conversion from 0-indexed half-open BED format | |
| #' to 1-indexed closed GenomicRanges format | |
| #' | |
| #' @param bed_df Data frame with chr, start, end columns in BED format (0-indexed, half-open) | |
| #' @param zero_indexed Logical, whether input is 0-indexed (default: TRUE) | |
| #' @return GRanges object | |
| bed_to_granges <- function(bed_df, zero_indexed = TRUE) { | |
| if (!all(c("chr", "start", "end") %in% names(bed_df))) { | |
| stop("bed_df must have columns: chr, start, end") | |
| } | |
| # Convert from 0-indexed half-open [start, end) to 1-indexed closed [start, end] | |
| if (zero_indexed) { | |
| gr_start <- bed_df$start + 1 | |
| gr_end <- bed_df$end | |
| } else { | |
| gr_start <- bed_df$start | |
| gr_end <- bed_df$end | |
| } | |
| # Create GRanges object (strand-agnostic for calling cards) | |
| gr <- GRanges( | |
| seqnames = bed_df$chr, | |
| ranges = IRanges(start = gr_start, end = gr_end), | |
| strand = "*" | |
| ) | |
| # Add any additional metadata columns | |
| extra_cols <- setdiff(names(bed_df), c("chr", "start", "end", "strand")) | |
| if (length(extra_cols) > 0) { | |
| mcols(gr) <- bed_df[, extra_cols, drop = FALSE] | |
| } | |
| return(gr) | |
| } | |
| #' Deduplicate insertions in GRanges object | |
| #' | |
| #' For calling cards, if an insertion is found at the same coordinate, | |
| #' only one record is retained | |
| #' | |
| #' @param gr GRanges object | |
| #' @return Deduplicated GRanges object | |
| deduplicate_granges <- function(gr) { | |
| # Find unique ranges (ignores strand and metadata) | |
| unique_ranges <- !duplicated(granges(gr)) | |
| gr[unique_ranges] | |
| } | |
| #' Count overlaps between insertions and regions | |
| #' | |
| #' @param insertions_gr GRanges object with insertions | |
| #' @param regions_gr GRanges object with regions | |
| #' @param deduplicate Whether to deduplicate insertions (default: TRUE) | |
| #' @return Integer vector of overlap counts per region | |
| count_overlaps <- function(insertions_gr, regions_gr, deduplicate = TRUE) { | |
| # Deduplicate if requested | |
| if (deduplicate) { | |
| n_before <- length(insertions_gr) | |
| insertions_gr <- deduplicate_granges(insertions_gr) | |
| n_after <- length(insertions_gr) | |
| if (n_before != n_after) { | |
| message( | |
| " Deduplicated: ", n_before, " -> ", n_after, | |
| " (removed ", n_before - n_after, " duplicates)" | |
| ) | |
| } | |
| } | |
| # Count overlaps per region | |
| # countOverlaps returns an integer vector with one element per region | |
| counts <- countOverlaps(regions_gr, insertions_gr) | |
| return(counts) | |
| } | |
| # Main Analysis Function -------------------------------------------------- | |
| #' Call peaks/quantify regions using calling cards approach | |
| #' | |
| #' @param experiment_gr GRanges object with experiment insertions | |
| #' @param background_gr GRanges object with background insertions | |
| #' @param regions_gr GRanges object with regions to quantify | |
| #' @param deduplicate_experiment Whether to deduplicate experiment insertions (default: TRUE) | |
| #' @param pseudocount Pseudocount for calculations (default: 0.1) | |
| #' @return GRanges object with regions and statistics as metadata columns | |
| enrichment_analysis <- function(experiment_gr, | |
| background_gr, | |
| regions_gr, | |
| deduplicate_experiment = TRUE, | |
| pseudocount = 0.1) { | |
| message("Starting enrichment analysis...") | |
| # Validate inputs | |
| if (!inherits(experiment_gr, "GRanges")) { | |
| stop("experiment_gr must be a GRanges object") | |
| } | |
| if (!inherits(background_gr, "GRanges")) { | |
| stop("background_gr must be a GRanges object") | |
| } | |
| if (!inherits(regions_gr, "GRanges")) { | |
| stop("regions_gr must be a GRanges object") | |
| } | |
| # Count overlaps for experiment (with deduplication if requested) | |
| message("Counting experiment overlaps...") | |
| if (deduplicate_experiment) { | |
| message(" Deduplication: ON") | |
| } else { | |
| message(" Deduplication: OFF") | |
| } | |
| experiment_counts <- count_overlaps( | |
| experiment_gr, regions_gr, | |
| deduplicate = deduplicate_experiment | |
| ) | |
| # Count overlaps for background (never deduplicated) | |
| message("Counting background overlaps...") | |
| message(" Deduplication: OFF (background should not be deduplicated)") | |
| background_counts <- count_overlaps( | |
| background_gr, regions_gr, | |
| deduplicate = FALSE | |
| ) | |
| # Calculate total hops AFTER any deduplication | |
| if (deduplicate_experiment) { | |
| experiment_gr_dedup <- deduplicate_granges(experiment_gr) | |
| total_experiment_hops <- length(experiment_gr_dedup) | |
| } else { | |
| total_experiment_hops <- length(experiment_gr) | |
| } | |
| total_background_hops <- length(background_gr) | |
| message("Total experiment hops: ", total_experiment_hops) | |
| message("Total background hops: ", total_background_hops) | |
| if (total_experiment_hops == 0) { | |
| stop("Experiment data is empty") | |
| } | |
| if (total_background_hops == 0) { | |
| stop("Background data is empty") | |
| } | |
| # Add counts and totals as metadata columns | |
| mcols(regions_gr)$experiment_hops <- as.integer(experiment_counts) | |
| mcols(regions_gr)$background_hops <- as.integer(background_counts) | |
| mcols(regions_gr)$total_experiment_hops <- as.integer(total_experiment_hops) | |
| mcols(regions_gr)$total_background_hops <- as.integer(total_background_hops) | |
| # Calculate statistics | |
| message("Calculating enrichment scores...") | |
| mcols(regions_gr)$callingcards_enrichment <- calculate_enrichment( | |
| total_background_hops = total_background_hops, | |
| total_experiment_hops = total_experiment_hops, | |
| background_hops = background_counts, | |
| experiment_hops = experiment_counts, | |
| pseudocount = pseudocount | |
| ) | |
| message("Calculating Poisson p-values...") | |
| mcols(regions_gr)$poisson_pval <- calculate_poisson_pval( | |
| total_background_hops = total_background_hops, | |
| total_experiment_hops = total_experiment_hops, | |
| background_hops = background_counts, | |
| experiment_hops = experiment_counts, | |
| pseudocount = pseudocount | |
| ) | |
| message("Calculating log Poisson p-values...") | |
| mcols(regions_gr)$log_poisson_pval <- calculate_poisson_pval( | |
| total_background_hops = total_background_hops, | |
| total_experiment_hops = total_experiment_hops, | |
| background_hops = background_counts, | |
| experiment_hops = experiment_counts, | |
| pseudocount = pseudocount, | |
| log.p = TRUE | |
| ) | |
| message("Calculating hypergeometric p-values...") | |
| mcols(regions_gr)$hypergeometric_pval <- calculate_hypergeom_pval( | |
| total_background_hops = total_background_hops, | |
| total_experiment_hops = total_experiment_hops, | |
| background_hops = background_counts, | |
| experiment_hops = experiment_counts | |
| ) | |
| message("Calculating log hypergeometric p-values...") | |
| mcols(regions_gr)$log_hypergeometric_pval <- calculate_hypergeom_pval( | |
| total_background_hops = total_background_hops, | |
| total_experiment_hops = total_experiment_hops, | |
| background_hops = background_counts, | |
| experiment_hops = experiment_counts, | |
| log.p = TRUE | |
| ) | |
| # Calculate adjusted p-values | |
| message("Calculating adjusted p-values...") | |
| mcols(regions_gr)$poisson_qval <- p.adjust(mcols(regions_gr)$poisson_pval, method = "fdr") | |
| mcols(regions_gr)$hypergeometric_qval <- p.adjust(mcols(regions_gr)$hypergeometric_pval, method = "fdr") | |
| message("Analysis complete!") | |
| return(regions_gr) | |
| } | |
| # Example Usage ----------------------------------------------------------- | |
| PROMOTERS <- "intergenic" | |
| genomic_features <- arrow::read_parquet("~/projects/huggingface/yeast_genome_resources/brentlab_features.parquet") | |
| genome_map_replicate_ds <- arrow::open_dataset("~/projects/huggingface/callingcards/genome_map", unify_schemas = TRUE) | |
| genome_map_replicate_meta <- arrow::read_parquet("~/projects/huggingface/callingcards/genome_map_meta.parquet") | |
| background_gr <- read_tsv("~/projects/huggingface/callingcards/adh1_background_ucsc.qbed") |> | |
| mutate( | |
| id = "adh1_bg", | |
| score = scales::rescale(depth, to = c(1, 1000)) | |
| ) |> | |
| dplyr::select(chr, start, end, id, score, strand) |> | |
| bed_to_granges() | |
| regions_list <- list( | |
| yiming = read_tsv("~/projects/huggingface/yeast_genome_resources/yiming_promoters.bed", | |
| col_names = c("chr", "start", "end", "locus_tag", "score", "strand") | |
| ) %>% | |
| bed_to_granges(), | |
| mindel = arrow::read_parquet("~/projects/huggingface/yeast_genome_resources/mindel_promoters.parquet", | |
| col_names = c("chr", "start", "end", "locus_tag", "score", "strand") | |
| ) %>% | |
| bed_to_granges(), | |
| start_codon_500bp = rtracklayer::import("~/projects/huggingface/yeast_genome_resources/start_codon_500bp_upstream_promoters.bed"), | |
| intergenic = rtracklayer::import("~/projects/huggingface/yeast_genome_resources/intergenic_regions_5_1.bed") | |
| ) | |
| # rename name to target_locus_tag | |
| colnames(GenomicRanges::mcols(regions_list$yiming))[1] <- "target_locus_tag" | |
| colnames(GenomicRanges::mcols(regions_list$start_codon_500bp))[1] <- "target_locus_tag" | |
| intergenic_meta <- read_csv("~/projects/huggingface/yeast_genome_resources/intergenic_regions_metadata_5_1.csv") |> | |
| dplyr::select(ir_name, chr, start, end, feature_left, feature_right) |> | |
| pivot_longer(-c(ir_name, chr, start, end), names_to = "side", values_to = "target_locus_tag") |> | |
| left_join(dplyr::select(genomic_features, | |
| target_locus_tag = locus_tag, | |
| target_symbol = symbol, target_strand = strand, | |
| target_start = start, target_end = end | |
| )) |> | |
| filter(chr != "chrM") |> | |
| filter(!is.na(target_strand)) |> | |
| mutate(valid = (target_strand == "-" & target_end <= start) | | |
| (target_strand == "+" & target_start >= end)) |> | |
| filter(valid) | |
| regions_gr <- regions_list[[PROMOTERS]] | |
| # removes AAD6 and AAD16 from mindel. pseudogene no longer in annotations | |
| if (PROMOTERS != "intergenic") { | |
| # really just for mindel | |
| regions_gr <- regions_gr[!is.na(regions_gr$target_locus_tag)] | |
| } else if (PROMOTERS == "intergenic") { | |
| regions_gr <- regions_gr[regions_gr$name %in% unique(intergenic_meta$ir_name)] | |
| } | |
| library(parallel) | |
| n_cores <- 10 | |
| results_replicates <- mclapply( | |
| genome_map_replicate_meta$id, | |
| \(x) enrichment_analysis( | |
| experiment_gr = genome_map_replicate_ds |> | |
| filter(id == x) |> | |
| collect() |> | |
| dplyr::rename(score = depth) |> | |
| relocate(chr, start, end, id, score, strand) |> | |
| mutate(depth = scales::rescale(score, to = c(1, 1000))) |> | |
| bed_to_granges(), | |
| background_gr = background_gr, | |
| regions_gr = regions_gr, | |
| deduplicate_experiment = TRUE, | |
| pseudocount = 0.1 | |
| ), | |
| mc.cores = n_cores | |
| ) | |
| names(results_replicates) <- genome_map_replicate_meta$id | |
| if (PROMOTERS == "intergenic") { | |
| results_replicates_df <- bind_rows(map(results_replicates, as_tibble), .id = "genome_map_id") |> | |
| mutate(genome_map_id = as.integer(genome_map_id)) |> | |
| dplyr::rename(ir_name = name) |> | |
| left_join(dplyr::select(intergenic_meta, ir_name, target_locus_tag, target_symbol), | |
| relationship = "many-to-many" | |
| ) |> | |
| left_join(genome_map_replicate_meta, | |
| by = c("genome_map_id" = "id") | |
| ) |> | |
| select( | |
| genome_map_id, batch, | |
| target_locus_tag, target_symbol, ir_name, | |
| experiment_hops, background_hops, | |
| total_background_hops, total_experiment_hops, | |
| callingcards_enrichment, | |
| poisson_pval, log_poisson_pval, poisson_qval, | |
| hypergeometric_pval, log_hypergeometric_pval, hypergeometric_qval | |
| ) |> | |
| mutate(genome_map_id = as.integer(genome_map_id)) | |
| # arrow::write_dataset( | |
| # results_replicates_df, | |
| # path = "/home/chase/projects/huggingface/callingcards/annotated_feature_reprocess_intergenic", | |
| # format = "parquet", | |
| # partitioning = c("batch"), | |
| # existing_data_behavior = "overwrite", | |
| # compression = "zstd", | |
| # write_statistics = TRUE, | |
| # use_dictionary = c( | |
| # genome_map_id = TRUE | |
| # ) | |
| # ) | |
| } else if (PROMOTERS == "start_codon_500bp") { | |
| results_replicates_df <- bind_rows(map(results_replicates, as_tibble), .id = "genome_map_id") |> | |
| mutate(genome_map_id = as.integer(genome_map_id)) |> | |
| left_join(dplyr::select(genomic_features, target_locus_tag = locus_tag, target_symbol = symbol)) |> | |
| left_join(genome_map_replicate_meta, | |
| by = c("genome_map_id" = "id") | |
| ) |> | |
| select( | |
| genome_map_id, batch, | |
| target_locus_tag, target_symbol, | |
| experiment_hops, background_hops, | |
| total_background_hops, total_experiment_hops, | |
| callingcards_enrichment, | |
| poisson_pval, log_poisson_pval, poisson_qval, | |
| hypergeometric_pval, log_hypergeometric_pval, hypergeometric_qval | |
| ) |> | |
| mutate(genome_map_id = as.integer(genome_map_id)) | |
| # arrow::write_dataset( | |
| # results_replicates_df, | |
| # path = "/home/chase/projects/huggingface/callingcards/annotated_feature_reprocess_start_codon_500", | |
| # format = "parquet", | |
| # partitioning = c("batch"), | |
| # existing_data_behavior = "overwrite", | |
| # compression = "zstd", | |
| # write_statistics = TRUE, | |
| # use_dictionary = c( | |
| # genome_map_id = TRUE | |
| # ) | |
| # ) | |
| } | |
| # creating the combined data for a new set | |
| # NOTE: THERE IS AN ERROR HERE. FOR THE SINGLETONS, THE ID IS THE | |
| # ANNOTATED_FEATURE ID, NOT THE GENOME_MAP_ID | |
| # as_2026 <- arrow::read_parquet("~/projects/huggingface/callingcards/deprecated/2026_analysis_set.parquet") | |
| # annotated_feature_meta <- arrow::read_parquet("~/projects/huggingface/callingcards/annotated_feature_meta.parquet") | |
| # as_2026_singles <- as_2026 |> | |
| # filter(str_detect(gm_id, "-", negate = TRUE)) |> | |
| # dplyr::rename(id = gm_id) |> | |
| # mutate(id = as.numeric(id)) |> | |
| # left_join( | |
| # annotated_feature_meta |> | |
| # dplyr::select(id, gm_id = genome_map_id) | |
| # ) |> | |
| # dplyr::relocate(id, gm_id) |> | |
| # dplyr::select(-id) |> | |
| # mutate(gm_id = as.character(gm_id)) | |
| # | |
| # as_2026_revised <- as_2026 |> | |
| # filter(str_detect(gm_id, "-")) |> | |
| # bind_rows(as_2026_singles) | |
| # | |
| # arrow::write_parquet(as_2026_revised, "~/projects/huggingface/callingcards/2026_analysis_set.parquet") | |
| as_2026 <- arrow::read_parquet("~/projects/huggingface/callingcards/2026_analysis_set.parquet") | |
| passing_rep_gm_ids <- unlist(map(unique(as_2026$gm_id), str_split, "-", simplify = TRUE)) | |
| genome_map_replicate_meta_combined <- genome_map_replicate_meta |> | |
| filter(id %in% passing_rep_gm_ids) |> | |
| group_by(regulator_locus_tag, regulator_symbol, condition) |> | |
| mutate(id = as.character(id)) |> | |
| reframe(combined_id = if (n() > 1) paste(id, collapse = "-") else id) | |
| # # Run analysis with deduplication (default for calling cards) | |
| results_combined <- map(genome_map_replicate_meta_combined$combined_id, ~ { | |
| # get list of ids (might just be 1) | |
| ids <- as.integer(strsplit(.x, "-")[[1]]) | |
| ids <- ids[!is.na(ids)] | |
| # for each id, produce a GRanges | |
| combined_passing_set <- map(ids, ~ { | |
| genome_map_replicate_ds |> | |
| filter(id == .x) |> | |
| collect() |> | |
| dplyr::rename(score = depth) |> | |
| relocate(chr, start, end, id, score, strand) |> | |
| mutate(depth = scales::rescale(score, to = c(1, 1000))) |> | |
| bed_to_granges() |> | |
| deduplicate_granges() | |
| }) | |
| # combine the list into a single gr | |
| combined_gr <- do.call(c, combined_passing_set) | |
| enrichment_analysis( | |
| experiment_gr = combined_gr, | |
| background_gr = background_gr, | |
| regions_gr = regions_gr, | |
| deduplicate_experiment = FALSE, | |
| pseudocount = 0.1 | |
| ) | |
| }) | |
| names(results_combined) <- genome_map_replicate_meta_combined$combined_id | |
| if (PROMOTERS == "intergenic") { | |
| results_combined_analysis_df <- bind_rows(map(results_combined, as_tibble), .id = "combined_id") |> | |
| dplyr::rename(ir_name = name) |> | |
| left_join(dplyr::select(intergenic_meta, ir_name, target_locus_tag, target_symbol), | |
| relationship = "many-to-many" | |
| ) |> | |
| left_join(genome_map_replicate_meta_combined) |> | |
| select( | |
| gm_id = combined_id, | |
| regulator_locus_tag, regulator_symbol, condition, | |
| # ir_name names the intergenic region the target was scored over. The | |
| # replicate-level branch above keeps it; keeping it here too makes the | |
| # published output joinable back to intergenic_regions_metadata_5_1.csv, | |
| # which is what the datacard's `join_column: ir_name` claims. | |
| target_locus_tag, target_symbol, ir_name, | |
| experiment_hops, background_hops, | |
| total_background_hops, total_experiment_hops, | |
| callingcards_enrichment, | |
| poisson_pval, log_poisson_pval, poisson_qval, | |
| hypergeometric_pval, log_hypergeometric_pval, hypergeometric_qval | |
| ) |> | |
| filter(condition == "standard") | |
| # results_combined_analysis_df |> | |
| # ungroup() |> | |
| # dplyr::select(-condition) |> | |
| # arrange(regulator_locus_tag) |> | |
| # arrow::write_parquet( | |
| # "/home/chase/projects/huggingface/callingcards/annotated_feature_reprocess_intergenic_analysis.parquet", | |
| # compression = "zstd", | |
| # write_statistics = TRUE, | |
| # use_dictionary = TRUE | |
| # ) | |
| } else if (PROMOTERS == "start_codon_500bp") { | |
| results_combined_analysis_df <- bind_rows(map(results_combined, as_tibble), .id = "combined_id") |> | |
| left_join(dplyr::select(genomic_features, target_locus_tag = locus_tag, target_symbol = symbol)) |> | |
| left_join(genome_map_replicate_meta_combined) |> | |
| select( | |
| gm_id = combined_id, | |
| regulator_locus_tag, regulator_symbol, condition, | |
| target_locus_tag, target_symbol, experiment_hops, background_hops, | |
| total_background_hops, total_experiment_hops, | |
| callingcards_enrichment, | |
| poisson_pval, log_poisson_pval, poisson_qval, | |
| hypergeometric_pval, log_hypergeometric_pval, hypergeometric_qval | |
| ) |> | |
| filter(condition == "standard") | |
| # results_combined_analysis_df |> | |
| # ungroup() |> | |
| # dplyr::select(-condition) |> | |
| # arrange(regulator_locus_tag) |> | |
| # arrow::write_parquet( | |
| # "/home/chase/projects/huggingface/callingcards/annotated_feature_reprocess_start_codon_500bp_analysis.parquet", | |
| # compression = "zstd", | |
| # write_statistics = TRUE, | |
| # use_dictionary = TRUE | |
| # ) | |
| } else if (PROMOTERS == "yiming") { | |
| results_combined_analysis_df <- bind_rows(map(results_combined, as_tibble), .id = "combined_id") |> | |
| left_join(dplyr::select(genomic_features, target_locus_tag = locus_tag, target_symbol = symbol)) |> | |
| left_join(genome_map_replicate_meta_combined) |> | |
| select( | |
| gm_id = combined_id, | |
| regulator_locus_tag, regulator_symbol, condition, | |
| target_locus_tag, target_symbol, experiment_hops, background_hops, | |
| total_background_hops, total_experiment_hops, | |
| callingcards_enrichment, | |
| poisson_pval, log_poisson_pval, poisson_qval, | |
| hypergeometric_pval, log_hypergeometric_pval, hypergeometric_qval | |
| ) |> | |
| filter(condition == "standard") | |
| out <- results_combined_analysis_df |> | |
| ungroup() |> | |
| dplyr::select(-condition) |> | |
| arrange(regulator_locus_tag) | |
| # out |> | |
| # arrow::write_parquet( | |
| # "/home/chase/projects/huggingface/callingcards/annotated_feature_reprocess_yiming_analysis.parquet", | |
| # compression = "zstd", | |
| # write_statistics = TRUE, | |
| # use_dictionary = TRUE | |
| # ) | |
| } else if (PROMOTERS == "mindel") { | |
| results_combined_analysis_df <- bind_rows(map(results_combined, as_tibble), .id = "combined_id") |> | |
| left_join(dplyr::select(genomic_features, target_locus_tag = locus_tag, target_symbol = symbol)) |> | |
| left_join(genome_map_replicate_meta_combined) |> | |
| select( | |
| gm_id = combined_id, | |
| regulator_locus_tag, regulator_symbol, condition, | |
| target_locus_tag, target_symbol, experiment_hops, background_hops, | |
| total_background_hops, total_experiment_hops, | |
| callingcards_enrichment, | |
| poisson_pval, log_poisson_pval, poisson_qval, | |
| hypergeometric_pval, log_hypergeometric_pval, hypergeometric_qval | |
| ) |> | |
| filter(condition == "standard") | |
| out <- results_combined_analysis_df |> | |
| ungroup() |> | |
| dplyr::select(-condition) |> | |
| arrange(regulator_locus_tag) | |
| # out |> | |
| # arrow::write_parquet( | |
| # "/home/chase/projects/huggingface/callingcards/annotated_feature_reprocess_mindel_analysis.parquet", | |
| # compression = "zstd", | |
| # write_statistics = TRUE, | |
| # use_dictionary = TRUE | |
| # ) | |
| } | |