Latent Factor Modeling Reveals Unexpected Spatial Heterogeneity in Human Alzheimer's Disease Brain Transcriptomes.
The 3 matches
- [1] § Methods › Single-sample GSEA ↔ supplementary/4.ST_WM_GSVA.qmd, lines 232–257 · score 0.61 · WM signature, ssGSEA, WM score, GSVA, RF, ROSMAP
- [2] § Methods › Random forest classifier ↔ supplementary/2.RF_score.qmd, lines 66–85 · score 0.53 · OOB accuracy, random forest, Model, MAP
- [3] § Methods › Discovery of the latent factor ↔ supplementary/1.DASC_multipleRun.qmd, lines 188–272 · score 0.51 · adjusted Rand, Pairwise, sensitivity, ranks, DASC, cluster
Paper
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The authors' code
Quarto · 378 lines · 12 KB · MIT · 1 match
- Calculate the correlation between RF classification probability for cluster 2 and WM scores using enriched WM siganture genes from supplementary table S4B, with those having t stats > 10
- ```{r setup}
- rm(list=ls())
- library(readxl)
- library(GSVA)
- library(ggplot2)
- library(dplyr)
- outDir <- "/home/zhijiany/workdir/2026/0402_AD_PaperReviews/out/4.ST_WM_GSVA"
- dir.create(outDir, recursive = TRUE, showWarnings = FALSE)
- suppFile <- "/home/zhijiany/workdir/2026/0402_AD_PaperReviews/data/supplementary_table.xlsx"
- countFile <- "/home/zhijiany/workdir/2026/0402_AD_PaperReviews/data/AD/ROSMAP_AD-N/ROSMAP_AMPAD_AD-N_counts.txt"
- rfScoreFile <- "/home/zhijiany/workdir/2026/0402_AD_PaperReviews/out/2.RF_score/ROSMAP_AMPAD_AD-N_meta_with_RF_scores.txt"
- rfValidationFile <- "/home/zhijiany/workdir/2026/0402_AD_PaperReviews/out/2.RF_score/validation_summary.csv"
- rfPerclassFile <- "/home/zhijiany/workdir/2026/0402_AD_PaperReviews/out/2.RF_score/perclass_metrics.csv"
- theme_set(theme_bw())
- # cluster colors: 1=green, 2=blue, 3=red
- clus_colors <- c("1" = "#00BA38", "2" = "#619CFF", "3" = "#F8766D")
- ```
- ## Helper functions
- ```{r helpers}
- run_ssgsea_safe <- function(expr_mat, gene_sets) {
- if ("ssgseaParam" %in% getNamespaceExports("GSVA")) {
- param <- GSVA::ssgseaParam(exprData = expr_mat, geneSets = gene_sets,
- alpha = 0.25,
- normalize = TRUE)
- ssgsea_res <- GSVA::gsva(param, verbose = FALSE)
- } else {
- ssgsea_res <- GSVA::gsva(expr = expr_mat, gset.idx.list = gene_sets,
- method = "ssgsea",
- ssgsea.norm = TRUE,
- verbose = FALSE)
- }
- as.numeric(ssgsea_res[1, ])
- }
- compute_correlations <- function(df, cohort_name) {
- tmp <- df |>
- filter(!is.na(RF_score_clus2), !is.na(WM_ssGSEA_score))
- if (nrow(tmp) < 3 ||
- length(unique(tmp$RF_score_clus2)) < 2 ||
- length(unique(tmp$WM_ssGSEA_score)) < 2) {
- return(data.frame(
- cohort = cohort_name,
- n = nrow(tmp),
- spearman_rho = NA_real_,
- spearman_p = NA_real_,
- pearson_r = NA_real_,
- pearson_p = NA_real_,
- stringsAsFactors = FALSE
- ))
- }
- spearman_test <- cor.test(tmp$RF_score_clus2, tmp$WM_ssGSEA_score,
- method = "spearman", exact = FALSE)
- pearson_test <- cor.test(tmp$RF_score_clus2, tmp$WM_ssGSEA_score,
- method = "pearson")
- data.frame(
- cohort = cohort_name,
- n = nrow(tmp),
- spearman_rho = unname(spearman_test$estimate),
- spearman_p = spearman_test$p.value,
- pearson_r = unname(pearson_test$estimate),
- pearson_p = pearson_test$p.value,
- stringsAsFactors = FALSE
- )
- }
- fmt_p <- function(x) {
- ifelse(x < 2.2e-16, "<2.2e-16", formatC(x, format = "e", digits = 2))
- }
- compute_boxplot_stats <- function(df) {
- plot_df <- df |>
- filter(!is.na(RF_predicted_clus), !is.na(WM_ssGSEA_score)) |>
- mutate(RF_predicted_clus = factor(RF_predicted_clus))
- kw_test <- kruskal.test(WM_ssGSEA_score ~ RF_predicted_clus, data = plot_df)
- group_levels <- levels(plot_df$RF_predicted_clus)
- pair_df <- utils::combn(group_levels, 2, simplify = FALSE) |>
- lapply(function(pair) {
- pair_data <- plot_df |>
- filter(RF_predicted_clus %in% pair)
- pair_test <- wilcox.test(WM_ssGSEA_score ~ RF_predicted_clus, data = pair_data,
- exact = FALSE)
- data.frame(
- group1 = pair[[1]],
- group2 = pair[[2]],
- p_value = pair_test$p.value,
- stringsAsFactors = FALSE
- )
- }) |>
- bind_rows() |>
- mutate(
- xmin = match(group1, group_levels),
- xmax = match(group2, group_levels)
- )
- y_max <- max(plot_df$WM_ssGSEA_score, na.rm = TRUE)
- y_min <- min(plot_df$WM_ssGSEA_score, na.rm = TRUE)
- y_range <- y_max - y_min
- if (!is.finite(y_range) || y_range == 0) {
- y_range <- max(abs(y_max), 1)
- }
- step_height <- 0.08 * y_range
- tick_height <- 0.015 * y_range
- label_offset <- 0.02 * y_range
- pair_df <- pair_df |>
- arrange(xmin, xmax) |>
- mutate(
- y = y_max + step_height * seq_len(n()),
- tick_y = y - tick_height,
- label_y = y + label_offset,
- label = paste0("Wilcoxon p=", fmt_p(p_value))
- )
- list(
- plot_df = plot_df,
- kw_label = paste0(
- "Kruskal-Wallis chi-squared=", sprintf("%.2f", unname(kw_test$statistic)),
- ", p=", fmt_p(kw_test$p.value)
- ),
- y_min = y_min,
- kw_y = y_max + step_height * (nrow(pair_df) + 1.2),
- y_upper = y_max + step_height * (nrow(pair_df) + 2.2),
- pair_df = pair_df
- )
- }
- ```
- ## Load RF-scored metadata and counts
- ```{r load-data}
- metaDf <- read.table(rfScoreFile, header = TRUE, sep = "\t",
- stringsAsFactors = FALSE, check.names = FALSE)
- metaDf$sample_id <- sub("^X", "", metaDf$newId)
- countDf <- read.table(countFile, header = TRUE, sep = "\t",
- stringsAsFactors = FALSE, check.names = FALSE)
- rownames(countDf) <- countDf[, 1]
- countDf <- as.matrix(countDf[, -1])
- storage.mode(countDf) <- "numeric"
- stopifnot(all(metaDf$sample_id %in% colnames(countDf)))
- metaDf <- metaDf[match(colnames(countDf), metaDf$sample_id), ]
- stopifnot(all(metaDf$sample_id == colnames(countDf)))
- cat("Samples in metadata:", nrow(metaDf), "\n")
- cat("Samples in counts:", ncol(countDf), "\n")
- print(table(metaDf$study))
- ```
- ## Load WM signature from Supplementary Table S4B
- ```{r signature}
- wmSigRaw <- read_excel(suppFile, sheet = "Table S4B")
- wmSigRaw <- as.data.frame(wmSigRaw, stringsAsFactors = FALSE)
- wmSigDf <- wmSigRaw |>
- select(ensembl, gene, t_stat_WM, p_value_WM, fdr_WM) |>
- filter(!is.na(ensembl), !is.na(t_stat_WM), t_stat_WM > 10) |>
- arrange(desc(t_stat_WM))
- wmGenesAll <- unique(wmSigDf$ensembl)
- wmGenesPresent <- intersect(wmGenesAll, rownames(countDf))
- wmGenesMissing <- setdiff(wmGenesAll, rownames(countDf))
- cat("WM signature genes (t_stat_WM > 10):", length(wmGenesAll), "\n")
- cat("Present in counts:", length(wmGenesPresent), "\n")
- cat("Missing from counts:", length(wmGenesMissing), "\n")
- stopifnot(length(wmGenesAll) == 302)
- stopifnot(length(wmGenesPresent) == 299)
- stopifnot(length(wmGenesMissing) == 3)
- wmSigPresentDf <- wmSigDf |> filter(ensembl %in% wmGenesPresent)
- wmSigMissingDf <- wmSigDf |> filter(ensembl %in% wmGenesMissing)
- write.table(wmSigPresentDf,
- file = file.path(outDir, "WM_signature_genes_tstat_gt10_present.txt"),
- sep = "\t", row.names = FALSE, quote = FALSE)
- write.table(wmSigMissingDf,
- file = file.path(outDir, "WM_signature_genes_tstat_gt10_missing.txt"),
- sep = "\t", row.names = FALSE, quote = FALSE)
- summaryLines <- c(
- paste("WM signature genes with t_stat_WM > 10:", length(wmGenesAll)),
- paste("Present in counts:", length(wmGenesPresent)),
- paste("Missing from counts:", length(wmGenesMissing)),
- paste("Missing genes:", paste(wmSigMissingDf$gene, collapse = ", "))
- )
- writeLines(summaryLines, con = file.path(outDir, "WM_signature_summary.txt"))
- wmSigMissingDf
- ```
- ## Prepare expression matrix for ssGSEA
- The ROSMAP matrix is already continuous normalized expression, so we use it
- directly for ssGSEA rather than re-running count-based normalization. Because
- ssGSEA is based on within-sample gene ranks, we score the combined matrix once
- across all samples and keep the default ssGSEA normalization enabled
- (`normalize = TRUE` or `ssgsea.norm = TRUE`) so WM enrichment scores are on a
- more comparable cross-sample scale for visualization and downstream summaries.
- ```{r expression-matrix}
- exprMat <- countDf
- stopifnot(identical(colnames(exprMat), metaDf$sample_id))
- stopifnot(identical(rownames(exprMat), rownames(countDf)))
- normSummaryDf <- metaDf |>
- count(study, name = "n_samples")
- write.table(normSummaryDf, file = file.path(outDir, "cohort_sample_counts.txt"),
- sep = "\t", row.names = FALSE, quote = FALSE)
- summary(as.numeric(exprMat[, 1:5]))
- ```
- ## Compute WM ssGSEA score per sample
- ```{r ssgsea-score}
- wmScores <- run_ssgsea_safe(
- expr_mat = exprMat,
- gene_sets = list(WM_signature = wmGenesPresent)
- )
- gsvaDf <- data.frame(
- sample_id = colnames(exprMat),
- WM_ssGSEA_score = wmScores,
- stringsAsFactors = FALSE
- )
- resDf <- metaDf |>
- left_join(gsvaDf, by = "sample_id")
- stopifnot(sum(is.na(resDf$WM_ssGSEA_score)) == 0)
- stopifnot(sum(is.na(resDf$RF_score_clus2)) == 0)
- write.table(resDf,
- file = file.path(outDir, "ROSMAP_AMPAD_AD-N_meta_with_RF_and_ssGSEA_scores.txt"),
- sep = "\t", row.names = FALSE, quote = FALSE)
- summary(resDf$WM_ssGSEA_score)
- ```
- ## Correlation between ssGSEA WM score and RF cluster 2 score
- ```{r correlations}
- corSummaryDf <- bind_rows(
- compute_correlations(resDf, "Pooled"),
- compute_correlations(filter(resDf, study == "MAP"), "MAP"),
- compute_correlations(filter(resDf, study == "ROS"), "ROS")
- )
- corSummaryDf$label <- paste0(
- corSummaryDf$cohort,
- ": rho=", sprintf("%.3f", corSummaryDf$spearman_rho),
- ", p=", fmt_p(corSummaryDf$spearman_p),
- ", n=", corSummaryDf$n
- )
- write.table(corSummaryDf,
- file = file.path(outDir, "WM_ssGSEA_vs_RF_score_correlations.txt"),
- sep = "\t", row.names = FALSE, quote = FALSE)
- corSummaryDf
- ```
- ## Reviewer-facing figures
- ```{r figures}
- pooled_label <- corSummaryDf$label[corSummaryDf$cohort == "Pooled"]
- p_scatter <- ggplot(resDf, aes(x = RF_score_clus2, y = WM_ssGSEA_score, color = study)) +
- geom_point(size = 2.2, alpha = 0.8) +
- geom_smooth(method = "lm", se = FALSE, linewidth = 0.8) +
- geom_smooth(
- data = resDf,
- aes(x = RF_score_clus2, y = WM_ssGSEA_score),
- inherit.aes = FALSE,
- method = "lm", se = FALSE, color = "black",
- linetype = "dashed", linewidth = 0.9
- ) +
- annotate("text", x = Inf, y = -Inf, label = pooled_label,
- hjust = 1.05, vjust = -0.8, size = 3.7) +
- labs(
- title = "WM ssGSEA score tracks RF cluster 2 score",
- subtitle = "Colored lines: cohort-specific fits; dashed black line: pooled fit",
- x = "RF_score_clus2",
- y = "WM ssGSEA score",
- color = "Cohort"
- )
- ggsave(file.path(outDir, "WM_ssGSEA_vs_RF_score_scatter.pdf"),
- p_scatter, width = 5.8, height = 5.8)
- p_dist <- ggplot(resDf, aes(x = study, y = WM_ssGSEA_score, fill = study)) +
- geom_violin(trim = FALSE, alpha = 0.5, color = NA) +
- geom_boxplot(width = 0.18, outlier.shape = NA, alpha = 0.85) +
- labs(
- title = "Distribution of WM ssGSEA scores by cohort",
- x = "Cohort",
- y = "WM ssGSEA score",
- fill = "Cohort"
- ) +
- guides(fill = "none")
- ggsave(file.path(outDir, "WM_ssGSEA_score_by_cohort.pdf"),
- p_dist, width = 5.5, height = 5)
- clusterStatList <- compute_boxplot_stats(resDf)
- p_cluster <- ggplot(clusterStatList$plot_df,
- aes(x = RF_predicted_clus, y = WM_ssGSEA_score,
- fill = RF_predicted_clus)) +
- geom_boxplot(alpha = 0.8, outlier.shape = NA) +
- geom_jitter(width = 0.12, alpha = 0.35, size = 1.2) +
- geom_segment(data = clusterStatList$pair_df,
- aes(x = xmin, xend = xmax, y = y, yend = y),
- inherit.aes = FALSE, linewidth = 0.5) +
- geom_segment(data = clusterStatList$pair_df,
- aes(x = xmin, xend = xmin, y = tick_y, yend = y),
- inherit.aes = FALSE, linewidth = 0.5) +
- geom_segment(data = clusterStatList$pair_df,
- aes(x = xmax, xend = xmax, y = tick_y, yend = y),
- inherit.aes = FALSE, linewidth = 0.5) +
- geom_text(data = clusterStatList$pair_df,
- aes(x = (xmin + xmax) / 2, y = label_y, label = label),
- inherit.aes = FALSE, size = 3.1) +
- annotate("text", x = 2, y = clusterStatList$kw_y, label = clusterStatList$kw_label,
- size = 3.3) +
- coord_cartesian(ylim = c(clusterStatList$y_min, clusterStatList$y_upper)) +
- scale_fill_manual(values = clus_colors) +
- labs(
- title = "WM ssGSEA score by RF-predicted cluster",
- x = "RF-predicted cluster",
- y = "WM ssGSEA score",
- fill = "RF cluster"
- )
- ggsave(file.path(outDir, "WM_ssGSEA_score_by_RF_predicted_cluster.pdf"),
- p_cluster, width = 6.2, height = 5)
- p_scatter
- ```
- ## Existing RF validation metrics
- ```{r rf-validation}
- rfValidationDf <- read.csv(rfValidationFile, stringsAsFactors = FALSE)
- rfPerclassDf <- read.csv(rfPerclassFile, stringsAsFactors = FALSE)
- write.table(rfValidationDf,
- file = file.path(outDir, "RF_validation_summary_reused.txt"),
- sep = "\t", row.names = FALSE, quote = FALSE)
- write.table(rfPerclassDf,
- file = file.path(outDir, "RF_perclass_metrics_reused.txt"),
- sep = "\t", row.names = FALSE, quote = FALSE)
- rfValidationDf
- ```
- ```{r}
- rfPerclassDf
- ```
4.ST_WM_GSVA.qmd at commit 9af7a5d, under MIT · at the source
Overview
- Department of Data Science and Artificial Intelligence, Al Hussein Technical University, Amman, Jordan
- Jan and Dan Duncan Neurologic Research Institute, Texas Children’s Hospital, Houston, TX 77030, USA
- Department of Pediatrics, Baylor College of Medicine, Houston, TX 77030, USA
- Center for Drug Discovery, Baylor College of Medicine, Houston, TX 77030, USA
- Department of Neuroscience, Baylor College of Medicine, Houston, TX 77030, USA
- Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX 77030, USA
- Department of Neurology, Baylor College of Medicine, Houston, TX 77030, USA
- Center for Alzheimer’s and Neurodegenerative Diseases, Baylor College of Medicine, Houston, TX 77030, USA
Abstract
Alzheimer’s disease is characterized by complex molecular and cellular heterogeneity, which complicates efforts to identify consistent biomarkers and therapeutic targets. To better characterize the heterogeneity, we applied latent factor modeling to RNA sequencing data from approximately 2,500 human Alzheimer’s disease brain samples, uncovering underlying patterns in gene expression. These transcriptional groups demonstrated unique gene expression profiles related to synaptic and neuronal pathways, vasculature development, and protein folding and antigen processing. Notably, this latent factor reflects variation in spatial sampling. Adjusting for the latent factor improved the identification of differentially expressed genes in disease samples. This finding suggests that spatial heterogeneity is a pervasive driver of transcriptomic variation and has important implications for future studies of Alzheimer’s disease and related neurological disorders.
Reproduced under the paper's license (CC BY), from the paper cited above.
Repository
Its files are read in the Code ↔ Paper reader above, with 3 matches between paragraphs and lines of code.
LiuzLab/AD_LatentFactor
9af7a5d1792672956fe3592eb5f264c2960eae16, 6 April 2026Availability: 1 check, the latest on 28 September 2026: the link answers
- 28 September 2026: the link answers
6 files
- supplementary/
1.DASC_multipleRun.qmd , Quarto, 272 lines, 1 match - supplementary/
2.RF_score.qmd , Quarto, 299 lines, 1 match - supplementary/
3.DEG_w_covariate.qmd , Quarto, 574 lines - supplementary/
4.ST_WM_GSVA.qmd , Quarto, 378 lines, 1 match - LICENSE, License, 21 lines
- README.md, Text, 2 lines
The paper's code and data availability statement is in the Data section.
Tracing map
Proposed by the machine: these links were found in the paper and verified at the source, without human review. The map will receive a Zenodo DOI once one of the paper's authors has validated it with their ORCID.
What the map holds:
- 1 repository of the authors' code, each at its verified commit, with its license and how the link was found in the paper;
- 4 scripts, each with its path and the digest of its content;
- 3 matches between paragraphs of the paper and lines of the code (method lexical-v1);
- neither the text of the paper nor the code itself.
Its JSON (tracing-map.json) is deposited on Zenodo with its DOI once the map is validated.
Data
No dataset and no data link were found in the paper.
Data Availability
Codes used in this study were available on github: https://
Reproduced under the paper's license (CC BY), from the paper cited above.
Versions
The history of this record: each version stored by the harvester or made by a correction of its authors or of the maintainers of its code, and what changed in its facts. The texts of the paper (its abstract, its availability statements) are not part of it; versions that changed only those are not listed.
Version 1, 28 September 2026: the first record
Recorded: type, language, journal, volume, issue, pages, dates, 13 authors, 2 funders, 35 references.
Cite
This paper
Al-Ouran, R., Liu, C., Wang, L., Yu, Z., Wan, Y.-W., Gu, C., Li, X., Cappuccio, G., Maletic-Savatic, M., Milosavljevic, A., Shulman, J. M., Chen, H., & Liu, Z. (2026). Latent Factor Modeling Reveals Unexpected Spatial Heterogeneity in Human Alzheimer's Disease Brain Transcriptomes. Computational and structural biotechnology journal, 35(1), 0108. https://
BibTeX
@article{alouran2026late
author = {Al-Ouran, Rami and Liu, Chaozhong and Wang, Linhua and Yu, Zhijian and Wan, Ying-Wooi and Gu, Chaohao and Li, Xiqi and Cappuccio, Gerarda and Maletic-Savatic, Mirjana and Milosavljevic, Aleksandar and Shulman, Joshua M. and Chen, Hu and Liu, Zhandong},
title = {{Latent Factor Modeling Reveals Unexpected Spatial Heterogeneity in Human Alzheimer's Disease Brain Transcriptomes}},
journal = {Computational and structural biotechnology journal},
year = {2026},
month = may,
volume = {35},
number = {1},
pages = {0108},
publisher = {AAAS Science Partner Journal Program},
issn = {2001-0370},
doi = {10.34133/
url = {https://
pmid = {42146901},
pmcid = {PMC13172580}
}
RIS
TY - JOUR
AU - Al-Ouran, Rami
AU - Liu, Chaozhong
AU - Wang, Linhua
AU - Yu, Zhijian
AU - Wan, Ying-Wooi
AU - Gu, Chaohao
AU - Li, Xiqi
AU - Cappuccio, Gerarda
AU - Maletic-Savatic, Mirjana
AU - Milosavljevic, Aleksandar
AU - Shulman, Joshua M.
AU - Chen, Hu
AU - Liu, Zhandong
TI - Latent Factor Modeling Reveals Unexpected Spatial Heterogeneity in Human Alzheimer's Disease Brain Transcriptomes
T2 - Computational and structural biotechnology journal
J2 - Comput Struct Biotechnol J
PY - 2026
DA - 2026/
VL - 35
IS - 1
SP - 0108
SN - 2001-0370
PB - AAAS Science Partner Journal Program
DO - 10.34133/
UR - https://
LA - en
ER -
CSL-JSON
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