{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T06:07:41Z","timestamp":1789106861554,"version":"build-2803163510"},"reference-count":40,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T00:00:00Z","timestamp":1789084800000},"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. Cell Dev. Biol."],"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Hepatocellular carcinoma (HCC) carries dismal prognosis, and the tumor immune microenvironment (TIME) critically determines the efficacy of SBRT-based radio-immunotherapy; yet its single-cell architecture remains undefined.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      We analyzed scRNA-seq data (GEO: GSE149614; single patient HCC07, stage IIIB; normal liver, primary tumor, PVTT) using a standardized pipeline including dimensionality reduction, clustering, immune signature scoring, and checkpoint profiling (FDR &amp;lt;0.05).\n                      <jats:italic>In vitro<\/jats:italic>\n                      , HepG2 and Huh-7 cells (ATCC) were treated with M2-conditioned medium for RT-qPCR\/ELISA validation of immunosuppressive mediators, and irradiated (Cs-137; 0\u201312\u00a0Gy) to assess radiation-induced immunogenic cell death modulation.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>scRNA-seq identified six major cell types across 25 clusters: myeloid cells expanded from 22.4% (normal) to 46.5% (tumor), while T\/NK cells declined from 66.5% to 20.0%. SPP1, CXCL8, APOE, TGF-\u03b21, IL-10, and VEGFA were upregulated in tumor\/PVTT, with multi-checkpoint T cell exhaustion (PDCD1, LAG3, TIGIT, CTLA4) dominating tumor-infiltrating lymphocytes. RT-qPCR\/ELISA confirmed M2-conditioned medium upregulated SPP1 (\u223c3.4-fold) and CXCL8 (\u223c4.1-fold; p &amp;lt; 0.001) in Hepatocellular carcinoma cells, and attenuated radiation-induced calreticulin exposure by \u223c40% (p &amp;lt; 0.05).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>The HCC TIME is profoundly immunosuppressive, driven by myeloid dominance and multi-checkpoint T cell exhaustion. The immunosuppressive microenvironment attenuates SBRT-induced ICD, providing direct mechanistic rationale for combining macrophage reprogramming, dual checkpoint blockade, and anti-angiogenic strategies with SBRT.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fcell.2026.1928725","type":"journal-article","created":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T05:29:44Z","timestamp":1789104584000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Single-cell transcriptomic characterization of the immunosuppressive tumor immune microenvironment in hepatocellular carcinoma: implications for SBRT-based radio-immunotherapy"],"prefix":"10.3389","volume":"14","author":[{"given":"Xiaofei","family":"Zhang","sequence":"first","affiliation":[{"name":"The Second Affiliated Hospital, Guangxi University of Chinese Medicine","place":["Nanning, Guangxi, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaohan","family":"Ma","sequence":"additional","affiliation":[{"name":"Graduate School, Guangxi University of Chinese Medicine","place":["Nanning, Guangxi, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sheng","family":"Chen","sequence":"additional","affiliation":[{"name":"Graduate School, Guangxi University of Chinese Medicine","place":["Nanning, Guangxi, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuanjun","family":"Mo","sequence":"additional","affiliation":[{"name":"The Second Affiliated Hospital, Guangxi University of Chinese Medicine","place":["Nanning, Guangxi, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuqun","family":"Lu","sequence":"additional","affiliation":[{"name":"The Second Affiliated Hospital, Guangxi University of Chinese Medicine","place":["Nanning, Guangxi, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Youke","family":"Xie","sequence":"additional","affiliation":[{"name":"The Second Affiliated Hospital, Guangxi University of Chinese Medicine","place":["Nanning, Guangxi, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xudong","family":"Liu","sequence":"additional","affiliation":[{"name":"The Second Affiliated Hospital, Guangxi University of Chinese Medicine","place":["Nanning, Guangxi, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Encun","family":"Hou","sequence":"additional","affiliation":[{"name":"The Second Affiliated Hospital, Guangxi University of Chinese Medicine","place":["Nanning, Guangxi, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,9,11]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"EVIDoa2100070","DOI":"10.1056\/EVIDoa2100070","article-title":"Tremelimumab plus durvalumab in unresectable hepatocellular carcinoma","volume":"1","author":"Abou-Alfa","year":"2022","journal-title":"NEJM Evid."},{"key":"B2","doi-asserted-by":"publisher","first-page":"409","DOI":"10.1038\/nrc3958","article-title":"The tumour microenvironment after radiotherapy: mechanisms of resistance and recurrence","volume":"15","author":"Barker","year":"2015","journal-title":"Nat. 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