{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T11:09:06Z","timestamp":1787656146083,"version":"build-2736575974"},"reference-count":39,"publisher":"BMJ","issue":"12","license":[{"start":{"date-parts":[[2025,12,30]],"date-time":"2025-12-30T00:00:00Z","timestamp":1767052800000},"content-version":"unspecified","delay-in-days":29,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"name":"Intramural Research Program, Center of Cancer Research, National Cancer Institute and NIH Clinical Center","award":["N\/A"],"award-info":[{"award-number":["N\/A"]}]},{"name":"Harold Amos Physician-Scientist Development Program Award","award":["N\/A"],"award-info":[{"award-number":["N\/A"]}]},{"name":"Alex\u2019s Lemonade Stand Center of Excellence","award":["N\/A"],"award-info":[{"award-number":["N\/A"]}]},{"name":"California Institute of Regenerative Medicine, UCSF Alpha Clinic Physician Scientist Research Fellowship Award","award":["N\/A"],"award-info":[{"award-number":["N\/A"]}]},{"name":"UCSF Medical Scientist Training Program","award":["T32GM141323"],"award-info":[{"award-number":["T32GM141323"]}]},{"name":"Contrato R\u00edo Hortega by the Instituto de Salud Carlos III","award":["CM24\/00094"],"award-info":[{"award-number":["CM24\/00094"]}]},{"name":"TERAV+ by RICORS\/TERAV","award":["RD24\/0014\/0003"],"award-info":[{"award-number":["RD24\/0014\/0003"]}]},{"name":"Baszucki Lymphoma Therapeutics Initiative","award":["N\/A"],"award-info":[{"award-number":["N\/A"]}]},{"name":"Dept. of Defense Congressionally Directed Medical Research Program Award","award":["W81XWH2210807"],"award-info":[{"award-number":["W81XWH2210807"]}]},{"name":"American Society of Hematology Research Training Award for Fellows","award":["N\/A"],"award-info":[{"award-number":["N\/A"]}]},{"name":"UCSF Helen Diller Family Comprehensive Cancer Center Preclinical Therapeutic Core Facility","award":["P30CA082103"],"award-info":[{"award-number":["P30CA082103"]}]},{"name":"Chan Zuckerberg Biohub Physician-Scientist Fellowship Program","award":["N\/A"],"award-info":[{"award-number":["N\/A"]}]},{"name":"National Institute of Health","award":["ZIA BC 011823"],"award-info":[{"award-number":["ZIA BC 011823"]}]},{"name":"UCSF Living Therapeutics Initiative","award":["N\/A"],"award-info":[{"award-number":["N\/A"]}]},{"name":"American Society of Transplantation and Cellular Therapy New Investigator Award","award":["N\/A"],"award-info":[{"award-number":["N\/A"]}]},{"name":"Multiple Myeloma Research Foundation Translational Accelerator Award","award":["N\/A"],"award-info":[{"award-number":["N\/A"]}]},{"name":"NIH","award":["T32AI007529"],"award-info":[{"award-number":["T32AI007529"]}]}],"content-domain":{"domain":["bmj.com"],"crossmark-restriction":true},"short-container-title":["J Immunother Cancer"],"accepted":{"date-parts":[[2025,11,25]]},"published-print":{"date-parts":[[2025,12]]},"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Chimeric antigen receptor (CAR) T-cell therapies are highly efficacious for several different hematologic cancers. However, for most CAR T targets it is observed that low surface antigen density on tumors can significantly reduce therapeutic efficacy. In this study, we explore this dynamic in the context of CD72, a surface antigen we recently found as a promising target for refractory B-cell cancers, but for which CD72 low antigen density can lead to therapeutic resistance in preclinical models.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Primary samples were accessed via institutional review board-approved protocols. Affinity-matured and humanized nanobody clones were previously described in Temple\n                      <jats:italic>et al.<\/jats:italic>\n                      (2023). CAR T cells were generated via lentiviral transduction. In vitro cytotoxicity assays were performed using luciferase-labeled cell lines. In vivo studies were performed using cell line-derived or patient-derived xenografts implanted in NOD\n                      <jats:italic>scid<\/jats:italic>\n                      gamma mice.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>We first confirmed ubiquitous CD72 expression across a range of primary B-cell non-Hodgkin lymphomas. We further found that after resistance to CD19-directed therapies, across both B-cell acute lymphoblastic leukemia (B-ALL) models and primary tumor samples, surface CD72 expression was largely preserved while CD22 expression was significantly diminished. Affinity maturation of a nanobody targeting CD72, when incorporated into CAR T cells, led to more effective elimination in vitro of isogenic models of CD72 low-expressing tumors. These results suggested that nanobody-based CAR T cells (nanoCARs) may exhibit a similar relationship between binder affinity, antigen expression, and efficacy as previously demonstrated only for single chain variable fragment-based CAR T cells. Surprisingly, however, this significantly improved in vitro efficacy only translated to modest in vivo survival benefit. As a parallel strategy to enhance CAR T function, we found that the small molecule bryostatin could also significantly increase CD72 surface antigen density on B-cell malignancy models. Structural modeling and biochemical analysis identified critical residues improving CD72 antigen recognition of our lead affinity-matured nanobody.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>Together, these findings support affinity-matured CD72 nanoCARs as a potential immunotherapy product for CD19-refractory B-cell cancers. Our results also suggest that for B-ALL in particular, CD72 may be a preferable second-line immunotherapy target over CD22.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1136\/jitc-2025-012013","type":"journal-article","created":{"date-parts":[[2025,12,30]],"date-time":"2025-12-30T19:00:19Z","timestamp":1767121219000},"page":"e012013","update-policy":"https:\/\/doi.org\/10.1136\/crossmarkpolicy","source":"Crossref","is-referenced-by-count":2,"title":["Affinity-matured CD72-targeting nanobody CAR T cells enhance elimination of antigen-low B-cell malignancies"],"prefix":"10.1136","volume":"13","clinical-trial-number":[{"clinical-trial-number":"nct03448393","registry":"10.18810\/clinical-trials-gov"},{"clinical-trial-number":"nct03827343","registry":"10.18810\/clinical-trials-gov"}],"author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3296-3261","authenticated-orcid":false,"given":"Adila","family":"Izgutdina","sequence":"first","affiliation":[{"name":"Department of Laboratory 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