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Integrated transcriptomic analysis of the temporal cortex identifies CRH and GAD2 as neuropathological markers and reveals altered immune microenvironment in Alzheimer’s disease
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  • Published: 25 February 2026

Integrated transcriptomic analysis of the temporal cortex identifies CRH and GAD2 as neuropathological markers and reveals altered immune microenvironment in Alzheimer’s disease

  • Pan Liu1 na1,
  • ChengLong Huang1 na1,
  • Lin Lu2,
  • Yi Lin Pang3 &
  • …
  • ZhaoYang Huang1 

Scientific Reports , Article number:  (2026) Cite this article

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We are providing an unedited version of this manuscript to give early access to its findings. Before final publication, the manuscript will undergo further editing. Please note there may be errors present which affect the content, and all legal disclaimers apply.

Subjects

  • Biomarkers
  • Computational biology and bioinformatics
  • Immunology
  • Neurology
  • Neuroscience

Abstract

Alzheimer’s disease (AD) is a progressive neurodegenerative disorder characterized by complex neuroimmune interactions. Identifying reliable neuropathological markers and understanding immune cell infiltration in the brain are essential for improving our understanding of AD pathology. We integrated four temporal cortex gene expression datasets from the GEO database (GSE36980, GSE37263, GSE118553, GSE122063). Differentially expressed genes (DEGs) were identified using RobustRankAggreg (RRA) and batch correction. Functional enrichment was analyzed via GO and KEGG, and hub genes were identified through protein–protein interaction networks and comparative intersection analysis. Diagnostic performance was evaluated using ROC curves, and immune cell infiltration was profiled with CIBERSORT, with significant immune subsets identified via Wilcoxon tests and LASSO regression. Analysis revealed 98 robust DEGs, prominently enriched in pathways related to synaptic transmission and neuroactive ligand-receptor interactions. Two hub genes, CRH and GAD2, were identified and validated as being significantly downregulated in AD. ROC analysis affirmed their high discriminatory value (AUC ≥ 0.7), with a combined model demonstrating good performance. Immune infiltration profiling in the AD temporal cortex uncovered significant alterations in six immune cell populations: M2 macrophages, activated dendritic cells, and resting mast cells were increased, while plasma cells, regulatory T cells (Tregs), and activated NK cells were decreased. However, no significant correlation was found between the expression of CRH/GAD2 and these immune cell alterations. CRH and GAD2 are potential neuropathological markers for AD. The distinct immune infiltration patterns observed highlight the involvement of both innate and adaptive immunity in AD pathogenesis, offering new insights for understanding AD pathology and informing future therapeutic strategies. The lack of direct correlation suggests that neuronal gene dysregulation and immune alterations may represent parallel or independently regulated pathological dimensions in AD.

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Data availability

The gene expression datasets used in this study are publicly available from the GEO database (accession numbers: GSE36980, GSE37263, GSE118553, GSE122063, and GSE132903). The code supporting the findings is available at: https://github.com/caoyezi/analysis-code. Processed data are available from the corresponding author upon reasonable request.

Abbreviations

AD:

Alzheimer’s disease

RRA:

RobustRankAggreg

Tregs:

Regulatory T cells

NK:

Natural killer

GO:

Gene ontology

KEGG:

Kyoto encyclopedia of genes and genomes

ROC:

Receiver operating characteristic

RMA:

Robust multiarray average

PPI:

Protein–protein interaction

AUC:

The area under the ROC curve

Ct:

The threshold cycle

SEM:

Standard error of the mean

CSF:

Cerebrospinal fluid

PET:

Positron emission tomography

CRH:

Corticotropin-releasing hormone

DLPFC:

Dorsolateral prefrontal cortex

BWS:

Bu-Wang San

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Acknowledgements

We extend our profound gratitude to the patients, their families, and caregivers for their invaluable participation and trust in this study.

Funding

This work was supported by the National Natural Science Foundation of China (Grant 82302026).

Author information

Author notes

  1. Pan Liu and ChengLong Huang contributed equally to this work.

Authors and Affiliations

  1. Department of Clinical Laboratory, Renmin Hospital of Wuhan University, Wuhan, 430060, Hubei, China

    Pan Liu, ChengLong Huang & ZhaoYang Huang

  2. Department of Orthopedics, Renmin Hospital of Wuhan University, Wuhan, China

    Lin Lu

  3. College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha, 410128, Hunan, China

    Yi Lin Pang

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Contributions

Pan Liu and Chenglong Huang contributed equally as co-first authors. Their contributions included: conceptualization and study design, data collection, software utilization and formal analysis, methodology implementation and validation, and writing—original draft preparation. Lin Lu contributions included: data collection and software utilization. Zhaoyang Huang and Yilin Pang contributed equally as co-corresponding authors. Their contributions included: supervision and project administration, funding acquisition, resource provision, writing—review and editing, and final approval of the version to be published. All authors have read and approved the final manuscript and take full responsibility for the integrity and accuracy of all aspects of the work.

Corresponding authors

Correspondence to Yi Lin Pang or ZhaoYang Huang.

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Competing interests

The authors declare no competing interests.

Ethics approval and consent to participate

The Institutional Review Board of the Institutional Ethics Committee at Renmin Hospital of Wuhan University, granted approval for this study (approval No.2025K-K290). The research was conducted in compliance with the principles outlined in the Declaration of Helsinki. Informed consent for participation in this study was obtained from the next of kin or legal guardians of the postmortem tissue donors.

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Liu, P., Huang, C., Lu, L. et al. Integrated transcriptomic analysis of the temporal cortex identifies CRH and GAD2 as neuropathological markers and reveals altered immune microenvironment in Alzheimer’s disease. Sci Rep (2026). https://doi.org/10.1038/s41598-026-40762-6

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  • Received: 23 December 2025

  • Accepted: 16 February 2026

  • Published: 25 February 2026

  • DOI: https://doi.org/10.1038/s41598-026-40762-6

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Keywords

  • Alzheimer disease
  • Diagnostic biomarkers
  • Immune cell infiltration
  • Bioinformatics
  • CRH
  • GAD2
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