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Single-cell transcriptomic analysis reveals immune remodeling in bone marrow during aged sepsis

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Aging substantially increases susceptibility to sepsis, yet the underlying mechanisms of immune dysfunction in the elderly remain incompletely understood. Polymicrobial sepsis was induced in young and aged male mice via cecal ligation and puncture (CLP). Bone marrow from four groups (Y-Sham, n = 2; Y-CLP, n = 3; A-Sham, n = 2; A-CLP, n = 3) was analyzed by single-cell RNA sequencing and intercellular communication inferred via CellChat. Age-related immune changes were evaluated, and Transwell assays were used to assess myeloid cell migration. Aged septic mice showed worsened organ damage and systemic inflammation, with reduced adaptive (18.7% vs. 41.3%) and increased innate immunity (58.8% vs. 37.7%, A-CLP vs. Y-CLP). Neutrophil chemotaxis was impaired; monocytes and macrophages adopted a hyperinflammatory yet functionally exhausted phenotype; dendritic cells showed increased antigen presentation with diminished mobility; B cell maturation was disrupted with regression to earlier developmental stages; and T cells shifted toward stress-responsive and regulatory programs. We identified a cluster-specific expansion of HSCs in aged sepsis (39.8% vs. 31.2% in A-CLP vs. Y-CLP) with impaired Lgals9–Cd44-mediated intercellular communication. Myeloid cell migration was impaired in A-CLP but partially restored by Lgals9–Cd44 activation. This study presents a comprehensive single-cell map of bone marrow immune dysfunction in aged sepsis and identifies impaired HSC–myeloid communication as a critical mechanism driving immune failure. Therapeutically targeting the Lgals9–Cd44 axis may restore immune coordination in elderly sepsis, although its clinical feasibility and safety remain to be validated.

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Data availability

In the meantime, the data are available from the corresponding author upon reasonable request.

Abbreviations

LPS :

Lipopolysaccharide

scRNA-seq :

Single-cell RNA sequencing

CLP :

Cecal ligation and puncture

PCA :

Principal Component Analysis

UMAP :

Uniform Manifold Approximation and Projection

SNN :

Shared nearest neighbor

DEGs :

Differentially expressed genes

GO :

Gene Ontology

IL-6 :

Interleukin-6

TNF-α :

Tumor Necrosis Factor-alpha

ELISA :

Enzyme-Linked Immunosorbent Assay

H&E :

Hematoxylin and Eosin

BMDNs :

Bone Marrow–Derived Neutrophils

BMDMs :

Bone Marrow–Derived Macrophages

DCs :

Dendritic cells

pDCs :

Plasmacytoid dendritic cells

HSCs :

Hematopoietic stem cells

ROS :

Reactive oxygen species

Treg :

Regulatory T cell

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Acknowledgements

We sincerely thank the staff of Hypers Biotechnologies Genomic Center for their technical support in single-cell RNA sequencing and data processing. We also acknowledge the support from colleagues at the Department of Critical Care Medicine and the Zhejiang Key Laboratory of Geriatrics for their valuable assistance during the study.

Funding

This work was supported by grants from Zhejiang Provincial Natural Science Foundation of China (LY21H150002).

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Author notes

  1. Yijia Lin and Zhiying Zhou contributed equally to this work.

Authors and Affiliations

  1. Affiliated Zhejiang Hospital, Zhejiang University School of Medicine, No. 1229 Gudun Road, Hangzhou, 310030, Zhejiang, China

    Yijia Lin & Zhiying Zhou

  2. Zhejiang University School of Medicine, No. 866 Yuhangtang Road, Hangzhou, 310058, Zhejiang, China

    Yijia Lin & Zhiying Zhou

  3. The Second School of Clinical Medicine, Zhejiang Chinese Medicine University, No. 548 Binwen Road, Hangzhou, 310053, Zhejiang, China

    Mingkun Yang

  4. Zhejiang Key Laboratory of Geriatrics and Geriatrics Institute of Zhejiang Province, Zhejiang Hospital, No. 1229 Gudun Road, Hangzhou, 310030, Zhejiang, China

    Yingzheng Weng

  5. Zhejiang Key Laboratory of Geriatrics and Geriatrics Institute of Zhejiang Province, Zhejiang Hospital, No. 1229 Gudun Road, Hangzhou, 310030, Zhejiang, China

    Zhouxin Yang

  6. Affiliated Zhejiang Hospital, Zhejiang University School of Medicine, No. 1229 Gudun Road, Hangzhou, 310030, Zhejiang, China

    Jing Yan

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Contributions

YJL designed the study, performed the experiments, and analyzed the data. ZYZ assisted with data interpretation, statistical analysis, and manuscript drafting. MKY and YZW contributed to animal handling, sample preparation, and data collection. JY supervised the study, provided critical care expertise, and revised the manuscript. ZXY provided conceptual guidance, contributed to study design, and oversaw the bioinformatics analysis. All authors read and approved the final manuscript.

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Correspondence to Zhouxin Yang or Jing Yan.

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Animal studies were approved by the the Zhejiang Experimental Animal Center (SCXK 2024–0002).

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Lin, Y., Zhou, Z., Yang, M. et al. Single-cell transcriptomic analysis reveals immune remodeling in bone marrow during aged sepsis. GeroScience (2025). https://doi.org/10.1007/s11357-025-01966-2

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