{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,12]],"date-time":"2026-09-12T06:10:20Z","timestamp":1789193420016,"version":"build-2803163510"},"reference-count":94,"publisher":"BMJ","issue":"11","license":[{"start":{"date-parts":[[2025,11,19]],"date-time":"2025-11-19T00:00:00Z","timestamp":1763510400000},"content-version":"unspecified","delay-in-days":18,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"name":"CAS Lamvac (Guangzhou) Biomedical Technology Co., Ltd","award":["N\/A."],"award-info":[{"award-number":["N\/A."]}]}],"content-domain":{"domain":["bmj.com"],"crossmark-restriction":true},"short-container-title":["J Immunother Cancer"],"accepted":{"date-parts":[[2025,10,26]]},"published-print":{"date-parts":[[2025,11]]},"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      While CD19-targeting chimeric antigen receptor T-cell (CAR-T) therapies have shown efficacy in B-cell hematological malignancies, treatment of antigen-negative or relapsed tumors remains challenging. The recombinant malaria protein VAR2CSA (rVAR2) binds specifically to placental-like chondroitin sulfate A (\n                      <jats:italic>pl<\/jats:italic>\n                      -CSA), which is expressed on many cancer types, enabling its use in various tumor-targeting platforms. However, rVAR2-based CAR-T strategies are limited by poor protein thermostability, short serum half-life, and safety concerns related to irreversible CAR activation. To address these limitations, we developed a navigable CAR-T cell system that uses an rVAR2-based navigator to direct CAR-T cells to\n                      <jats:italic>pl<\/jats:italic>\n                      -CSA\n                      <jats:sup>+<\/jats:sup>\n                      tumors.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      We overexpressed and purified rVAR2 and three catenation proteins in\n                      <jats:italic>Escherichia coli<\/jats:italic>\n                      , then produced and epitope-mapped the anti-rVAR2 monoclonal antibody 5H4VAR2omab following murine immunization. Using its single-chain fragment variable, we constructed a third-generation CAR-T with CD28 and 4-1BB co-stimulatory domains. An interlocking rVAR2 catenane, AXVB, was identified via orthogonal screening and employed as a navigator for directing CAR-T cells to\n                      <jats:italic>pl<\/jats:italic>\n                      -CSA\n                      <jats:sup>+<\/jats:sup>\n                      tumors. The antitumor efficacy of this navigable CAR-T cell system was evaluated in vitro and in hematologic cancer cell line-derived xenograft models.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      We developed an orthogonal CAR-T cell system that employs a navigator protein (rVAR2 or AXVB) to target\n                      <jats:italic>pl<\/jats:italic>\n                      -CSA\n                      <jats:sup>+<\/jats:sup>\n                      tumors. Through a reversible switch mechanism, this system enables indirect tumor recognition by CAR-T cells, thereby improving the safety profile of conventional CAR-T therapies. Compared with the rVAR2 monomer, the AXVB multimer exhibited enhanced thermostability and target affinity. Both in vitro and in vivo, AXVB-(navigation)-CAR T cells potently eliminated CD19-positive and CD19-negative\n                      <jats:italic>pl<\/jats:italic>\n                      -CSA\n                      <jats:sup>+<\/jats:sup>\n                      tumor cells and significantly prolonged survival in tumor-bearing mice.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>Protein catenation improved the thermostability and functional activity of rVAR2. In a murine model of hematologic malignancy, CAR-T cells utilizing the rVAR2 catenane (AXVB) as a targeting module exhibited superior antitumor activity.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1136\/jitc-2025-012618","type":"journal-article","created":{"date-parts":[[2025,11,19]],"date-time":"2025-11-19T17:40:19Z","timestamp":1763574019000},"page":"e012618","update-policy":"https:\/\/doi.org\/10.1136\/crossmarkpolicy","source":"Crossref","is-referenced-by-count":2,"title":["Protein catenation potentiates antitumor activity of malaria VAR2CSA navigation CAR-T cells in a mouse model of hematological malignancies"],"prefix":"10.1136","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9561-1899","authenticated-orcid":false,"given":"Wen","family":"Hu","sequence":"first","affiliation":[{"name":"CAS Lamvac (Guangzhou) Biomedical Technology Co., Ltd, Guangzhou, China"},{"name":"Guangzhou Lamgene Biomedical Technology Co., Ltd, Guangzhou, 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