{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,26]],"date-time":"2026-06-26T15:33:13Z","timestamp":1782487993079,"version":"3.54.5"},"reference-count":27,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,12,18]],"date-time":"2025-12-18T00:00:00Z","timestamp":1766016000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100017596","name":"Natural Science Basic Research Program of Shaanxi Province","doi-asserted-by":"publisher","award":["2023-JC-QN-0923, S2026-JC-YB-C-1708"],"award-info":[{"award-number":["2023-JC-QN-0923, S2026-JC-YB-C-1708"]}],"id":[{"id":"10.13039\/501100017596","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>Microsatellite stable colorectal cancer (MSS-CRC) is referred to as an immune desert-type tumor. Emerging studies have demonstrated that inhibition of the Wnt\/\u03b2-catenin pathway can enhance the anti-tumor immune response. This research focuses on synthesizing the human serum albumin (HSA) -based Wnt inhibitor GHSACA and assessing its internalization pathway and therapeutic effectiveness in conjunction with PD-1 antibody for treating MSS-CRC.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Glucosamine was covalently conjugated to HSA via EDC\/NHS activation to generate GHSA, which was subsequently coupled with carnosic acid (CA) -Wnt pathway inhibitor, to synthesize GHSACA. For\n                      <jats:italic>in vitro<\/jats:italic>\n                      studies, LOX1- and CD44- deficient MC38 and CT26 colon cancer cell lines, validated by RT-qPCR, were used to investigate the cellular uptake mechanism and macropinocytic activity of GHSACA using flow cytometry. The effectiveness of GHSACA in targeting tumors\n                      <jats:italic>in vivo<\/jats:italic>\n                      was assessed in CT26 tumor-bearing BALB\/c mice via fluorescence imaging. Therapeutic efficacy was assessed in MSS-CRC patient-derived xenograft (PDX).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      RT-qPCR confirmed efficient knockdown of LOX1 and CD44 in both MC38 and CT26 cell lines. Cellular uptake assay demonstrated that GHSACA internalization is predominantly mediated by the LOX1-dependent macropinocytosis pathway.\n                      <jats:italic>In vivo<\/jats:italic>\n                      fluorescence imaging revealed sustained accumulation of GHSACA in tumor and rapid clearance from normal organs. In the PDX model, GHSACA monotherapy significantly suppressed tumor growth (TGI = 59.3%). The combination with PD-1 antibody (G&amp;amp;P group) resulted in further enhancement of antitumor efficacy (TGI = 87.9%). TUNEL assays showed the most pronounced induction of tumor cell apoptosis in the G&amp;amp;P group. Immunohistochemical analysis demonstrated that GHSACA suppressed the Wnt\/\u03b2-catenin signaling cascades, alongside with lower Ki67 expression. The G&amp;amp;P combination increased PD-L1 expression and significantly boosted Granzyme B-positive cytotoxic immune responses. Immunofluorescence double staining showed the highest infiltration of CD3\n                      <jats:sup>+<\/jats:sup>\n                      \/CD8\n                      <jats:sup>+<\/jats:sup>\n                      cytotoxic T lymphocytes of the G&amp;amp;P group, with a concurrent decrease in CD4\n                      <jats:sup>+<\/jats:sup>\n                      \/CD25\n                      <jats:sup>+<\/jats:sup>\n                      regulatory T cells. GHSACA was found to have favorable systemic biocompatibility and safety.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>GHSACA achieves efficient targeted cellular internalization via a macropinocytosis pathway regulated by the LOX1 receptor rather than the CD44 receptor, simultaneously inhibiting the Wnt\/\u03b2-catenin signaling while activating anti-tumor immune responses. It provides a highly promising translational therapeutic approach for overcoming immune resistance in MSS-CRC.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fimmu.2025.1737031","type":"journal-article","created":{"date-parts":[[2025,12,18]],"date-time":"2025-12-18T06:45:35Z","timestamp":1766040335000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["LOX1-mediated supramolecular self-assembly nanomedicine for microsatellite-stable colorectal cancer towards reactivating anti-tumor immunity"],"prefix":"10.3389","volume":"16","author":[{"given":"Xin","family":"Wei","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuyan","family":"Guo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianyuan","family":"Lei","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Na","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,12,18]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.3760\/cma.j.cn112152-20240308-00102","article-title":"Comparison analyses of global burden of colorectal cancer","volume":"47","author":"Li","year":"2025","journal-title":"Zhonghua Zhong Liu Za Zhi"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1200\/JCO.19.03296","article-title":"Regorafenib plus nivolumab in patients with advanced gastric or colorectal cancer: an open-label, dose-escalation, and dose-expansion phase ib trial (REGONIVO, EPOC1603)","volume":"38","author":"Fukuoka","year":"2020","journal-title":"J Clin Oncol"},{"key":"B3","doi-asserted-by":"publisher","first-page":"1729","DOI":"10.1016\/j.ccell.2024.08.019","article-title":"Fusobacterium nucleatum facilitates anti-PD-1 therapy in microsatellite stable colorectal cancer","volume":"42","author":"Wang","year":"2024","journal-title":"Cancer Cell"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2025.1587950","article-title":"Interplay between\u00a0tumor cells and immune cells of the colorectal cancer tumor microenvironment: Wnt\/\u03b2-catenin pathway","volume":"16","author":"Janeeh","year":"2025","journal-title":"Front Immunol"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1016\/j.bioactmat.2024.09.022","article-title":"Self-assembled PROTACs enable protein degradation to reprogram the tumor microenvironment for synergistically enhanced colorectal cancer immunotherapy","volume":"43","author":"Lu","year":"2025","journal-title":"Bioact Mater"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.7150\/thno.83377","article-title":"Targeting \u03b2-catenin and PD-L1 simultaneously by a racemic supramolecular peptide for the potent immunotherapy of hepatocellular carcinoma","volume":"13","author":"Zhou","year":"2023","journal-title":"Theranostics"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1038\/s41392-021-00762-6","article-title":"Wnt\/\u03b2-catenin signalling: function, biological mechanisms, and therapeutic opportunities","volume":"7","author":"Liu","year":"2022","journal-title":"Signal Transduct Target Ther"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2023.1274223","article-title":"Modulating \u03b2-catenin\/BCL9 interaction with cell-membrane-camouflaged carnosic acid to inhibit Wnt pathway and enhance tumor immune response","volume":"14","author":"Gao","year":"2023","journal-title":"Front Immunol"},{"key":"B9","doi-asserted-by":"publisher","first-page":"101412","DOI":"10.1016\/j.jpha.2025.101412","article-title":"Unlocking wnt\u2019s weak spot: glycosylated nanoalbumins to reignite immune responses in MSS-CRC","author":"Wei","year":"2025","journal-title":"J Pharm Anal"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1080\/21691401.2021.1884085","article-title":"CD44-targeted pH- responsive micelles for enhanced cellular internalization and intracellular on-demand release of doxorubicin","volume":"49","author":"Yang","year":"2021","journal-title":"Artif Cells Nanomed Biotechnol"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1080\/02648725.2023.2199477","article-title":"A comprehensive analysis of the expression and the prognosis for LOX-1 in multiple cancer types","volume":"40","author":"Wang","year":"2024","journal-title":"Biotechnol Genet Eng Rev"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1016\/j.bmcl.2024.129762","article-title":"The association between oxidized low-density lipoprotein and cancer: An emerging targeted therapeutic approach","volume":"106","author":"Moghadam","year":"2024","journal-title":"Bioorg Med Chem Lett"},{"key":"B13","doi-asserted-by":"publisher","first-page":"5107","DOI":"10.1038\/s41598-021-84492-3","article-title":"Lysyl oxidase engineered lipid nanovesicles for the treatment of triple negative breast cancer","volume":"11","author":"De Vita","year":"2021","journal-title":"Sci Rep"},{"key":"B14","doi-asserted-by":"publisher","first-page":"2456","DOI":"10.1016\/j.immuni.2025.07.013","article-title":"Inhibition of tumor cell macropinocytosis driver DHODH reverses immunosuppression and overcomes anti-PD1 resistance","volume":"58","author":"Wang","year":"2025","journal-title":"Immunity"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1021\/acsnano.5c08486","article-title":"Mito-specific \u201cTrojan horse\u201d Nanotherapy: synergistic antitumor immunotherapy via dual modulation mitochondrial metabolism and glycolysis","volume":"19","author":"Yang","year":"2025","journal-title":"ACS Nano"},{"key":"B16","doi-asserted-by":"publisher","first-page":"e2405758","DOI":"10.1002\/advs.202405758","article-title":"Targeting the NOTCH2\/ADAM10\/TCF7L2 axis-mediated transcriptional regulation of wnt pathway suppresses tumor growth and enhances chemosensitivity in colorectal cancer","volume":"12","author":"Xiang","year":"2025","journal-title":"Adv Sci (Weinh)"},{"key":"B17","doi-asserted-by":"publisher","first-page":"15","DOI":"10.1007\/s10238-023-01274-z","article-title":"WNT\/\u03b2-catenin regulatory roles on PD-(L)1 and immunotherapy responses","volume":"24","author":"Mortezaee","year":"2024","journal-title":"Clin Exp Med"},{"key":"B18","doi-asserted-by":"publisher","first-page":"101","DOI":"10.1186\/s13045-017-0471-6","article-title":"Wnt\/beta-catenin pathway: modulating anticancer immune response","volume":"10","author":"Pai","year":"2017","journal-title":"J Hematol Oncol"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-025-02597-w","article-title":"Turn cold tumors hot by reprogramming the tumor\u00a0microenvironment","volume":"43","author":"Yuan","year":"2025","journal-title":"Nat Biotechnol"},{"key":"B20","doi-asserted-by":"publisher","first-page":"eaau5240","DOI":"10.1126\/sciadv.aau5240","article-title":"Pharmacological inhibition of \u03b2-catenin\/BCL9 interaction overcomes resistance to immune checkpoint blockades by modulating T(reg) cells","volume":"5","author":"Feng","year":"2019","journal-title":"Sci Adv"},{"key":"B21","doi-asserted-by":"publisher","first-page":"1378","DOI":"10.1038\/s41591-025-03698-4","article-title":"PD-L1 and the dawn of modern cancer immunotherapy","volume":"31","author":"Chen","year":"2025","journal-title":"Nat Med"},{"key":"B22","doi-asserted-by":"publisher","first-page":"122","DOI":"10.1038\/s41467-023-44391-9","article-title":"FOXP3(+) regulatory T cell perturbation mediated by the IFN\u03b3-STAT1-IFITM3 feedback loop is essential for anti-tumor immunity","volume":"15","author":"Liu","year":"2024","journal-title":"Nat Commun"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2025.116302","article-title":"T cell-intrinsic cGAS-CTCF regulation maintains regulatory T cell development and function","volume":"44","author":"Li","year":"2025","journal-title":"Cell Rep"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2025.1502094","article-title":"Identification of Dinaciclib and Ganetespib as anti-inflammatory drugs using a novel HTP screening assay that targets IFN\u03b3-dependent PD-L1","volume":"16","author":"Hagbi-Levi","year":"2025","journal-title":"Front Immunol"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1038\/s41416-022-01960-x","article-title":"Dynamic host immunity and PD-L1\/PD-1 blockade efficacy: developments after \u201cIFN-\u03b3 from lymphocytes induces PD-L1 expression and promotes progression of ovarian cancer","volume":"128","author":"Abiko","year":"2023","journal-title":"Br J Can"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1038\/s41588-025-02077-6","article-title":"Alterations in PD-L1 succinylation shape anti-tumor immune responses in melanoma","volume":"57","author":"Liang","year":"2025","journal-title":"Nat Genet"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1007\/s00262-018-2283-0","article-title":"Enhanced anti-tumor efficacy of checkpoint inhibitors in combination with the histone deacetylase inhibitor Belinostat in a murine hepatocellular carcinoma model","volume":"68","author":"Llopiz","year":"2019","journal-title":"Cancer Immunol Immunother"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2025.1737031\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,18]],"date-time":"2025-12-18T06:45:38Z","timestamp":1766040338000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2025.1737031\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,18]]},"references-count":27,"alternative-id":["10.3389\/fimmu.2025.1737031"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2025.1737031","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,12,18]]},"article-number":"1737031"}}