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Linear-Mixed-Models-Recover-Immune-Pathway

R pipeline for linear mixed model–based recovery of immune pathway signals in γδ T cell RNA-seq.

Manuscript Title
Linear Mixed Models Recover Immune Pathway Signatures Masked by Inter-individual Variance in Sorted γδ T Cell RNA-seq: IFN-α and B Cell Activation as Correlates of HIV Neutralization Breadth

Author
Dohoon Kim
PromptGenix LLC
Corresponding author: dkim@promptgenix.org


Overview

This repository provides a unified R pipeline for transcriptomic and pathway-level analysis using linear mixed-effects models.
The workflow integrates differential expression, gene set enrichment, and validation across independent cohorts.


How to Run

Place the required input files in ~/Downloads, then run:

source("RV217_UNIFIED_PIPELINE_GitHub.R")

Input Data

  • final_metadata.csv
    Sample metadata (reconstructed from GSE271442 supplementary data)

  • GSE271442_Merged_with_Symbols.csv
    TPM expression matrix (31 MB) with gene symbols
    (unzip csv.zip before use)


Public Datasets

  • Primary dataset: GSE271442
  • Validation datasets: GSE124731, GSE24081

All datasets are publicly available through NCBI GEO.


Repository Contents

  • RV217_UNIFIED_PIPELINE_GitHub.R
    Compact reproducible version of the unified analysis pipeline and figure-generation code

Analysis Workflow

  • metadata harmonization
  • expression matrix processing and filtering
  • differential expression analysis (limma)
  • linear mixed-effects modeling (LMM)
  • pathway analysis (fGSEA, GSVA)
  • cross-cohort validation
  • manuscript figure generation

Output

The pipeline generates:

  • RV217_UNIFIED_WORKSPACE.RData (analysis objects)
  • publication-ready figures (PDF format)

Data Availability

The RNA-seq dataset used in this study (GSE271442) is publicly available in NCBI GEO:
https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE271442

Validation datasets (GSE124731 and GSE24081) are also publicly available in GEO.

Original data processing details are available in:
https://pmc.ncbi.nlm.nih.gov/articles/PMC11805418/


Citation

If you use this repository, please cite the associated manuscript and the original GEO datasets.

About

R code for "Linear Mixed Models Recover Immune Pathway Signatures Masked by Inter-individual Variance in Sorted γδ T Cell RNA-seq: IFN-α and B Cell Activation as Correlates of HIV Neutralization Breadth" paper

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