{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,5]],"date-time":"2026-06-05T12:05:07Z","timestamp":1780661107063,"version":"3.54.1"},"reference-count":21,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,5,22]],"date-time":"2026-05-22T00:00:00Z","timestamp":1779408000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Intervertebral disc degeneration (IDD) is increasingly viewed as an immune-perturbed and metabolically stressed niche rather than a purely mechanical or aging-related disorder. Mesenchymal stromal cells (MSCs) and their extracellular vesicles (EVs) may shape local immune signaling, yet MSC EV\u2013associated transcriptional programs and their immune-network context remain poorly defined at single-cell resolution.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      We analyzed single-cell RNA sequencing (scRNA-seq) data from GSE230809 (8 IDD and 3 control nucleus pulposus samples) with standard preprocessing and batch correction. We derived a transcriptome-inferred MSC EV\u2013associated program score by integrating complementary gene-set scoring strategies, stratified MSCs into EV-score\u2013high versus \u2013low states, and prioritized candidate regulators using a consensus LASSO\/SVM-RFE workflow. CellChat was used to infer ligand\u2013receptor communication, while pathway-level immunometabolic remodeling was assessed with GSVA\/GSEA and immune-signature correlation analyses. Selected genes were examined by RT\u2013qPCR in an\n                      <jats:italic>in vitro<\/jats:italic>\n                      inflammatory NP-cell model, and docking was performed to explore tractable compound\u2013target hypotheses.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>We resolved seven major cell populations and observed a marked shift toward higher EV-program scores in MSCs from IDD tissues. EV-score\u2013high MSCs showed increased incoming signaling and interaction potential in CellChat networks, consistent with an \u201cimmune cue\u2013responsive\u201d state. Machine-learning prioritization converged on five hub genes (AP2S1, CSTB, GSTP1, RPL28, and TSG101). The hub-gene program aligned with immune\u2013metabolic axes including IL6\/JAK\u2013STAT3 and interferon-related signaling, together with oxidative phosphorylation and ROS stress signatures, and was systematically associated with immune mediators (notably chemokines and checkpoint-related genes). AP2S1 and CSTB displayed dynamic expression along MSC pseudotime trajectories and were supported by RT\u2013qPCR under inflammatory stimulation. A hub-gene model distinguished IDD from control samples within the discovery cohort (AUC\u00a0=\u00a00.836). Docking suggested a plausible interaction between CSTB and ripasudil (\u22125.98 kcal\/mol).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>This study maps an MSC EV\u2013associated program at single-cell resolution and places it within an immune\u2013communication and immunometabolic framework in IDD. AP2S1 and CSTB are candidate nodes linking EV-program states to immune signaling and metabolic stress, providing candidate biomarkers and tractable intervention hypotheses.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fimmu.2026.1820174","type":"journal-article","created":{"date-parts":[[2026,5,22]],"date-time":"2026-05-22T05:01:43Z","timestamp":1779426103000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Single-cell extracellular vesicle-program scoring maps immunometabolic rewiring and immune crosstalk of mesenchymal stromal cells in intervertebral disc degeneration, prioritizing AP2S1 and CSTB"],"prefix":"10.3389","volume":"17","author":[{"given":"Fengyu","family":"Ma","sequence":"first","affiliation":[{"name":"Department of Orthopedics, The Second Affiliated Hospital of Soochow University","place":["Suzhou, Jiangsu, China"]},{"name":"Department of Spinal Surgery, People\u2019s Hospital of Rizhao","place":["Rizhao, Shandong, China"]},{"name":"Medical 3D Printing Center, Orthopedic Institute, Department of Orthopedic Surgery, The First Affiliated Hospital, School of Basic Medical Sciences, Interdisciplinary Innovation Center for Nanomedicine, MOE Key Laboratory of Geriatric Diseases and Immunology, Suzhou Medical College, Soochow University","place":["Suzhou, Jiangsu, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kelv","family":"Shen","sequence":"additional","affiliation":[{"name":"Department of Orthopedics, The Second Affiliated Hospital of Soochow University","place":["Suzhou, Jiangsu, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ziqiang","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Orthopedics, The Second Affiliated Hospital of Soochow University","place":["Suzhou, Jiangsu, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hanshi","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of Orthopedics, The Second Affiliated Hospital of Soochow University","place":["Suzhou, Jiangsu, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mingzhe","family":"Yu","sequence":"additional","affiliation":[{"name":"Department of Orthopedics, The Second Affiliated Hospital of Soochow University","place":["Suzhou, Jiangsu, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhendong","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Orthopedics, The Second Affiliated Hospital of Soochow University","place":["Suzhou, Jiangsu, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fengxuan","family":"Han","sequence":"additional","affiliation":[{"name":"Medical 3D Printing Center, Orthopedic Institute, Department of Orthopedic Surgery, The First 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