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However, alternate cytotoxic cell death mechanisms have been reported, potentially contributing to the overall anti-tumor activity. Although previous studies have highlighted a mechanism of caspase-independent cell death, this mechanism has yet to be well-characterized, thereby warranting further investigation to comprehensively understand the anti-tumor mechanism of CD47 blockade to facilitate optimal drug partner selection for combination therapy.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>The fully humanized anti-CD47 monoclonal antibodies, SRF231, magrolimab, as well as a mouse monoclonal anti-CD47 antibody, B6H12, were used. Multiple cell death mechanisms were evaluated including apoptosis, autophagy and necroptosis by using customized Hoechst\/Annexin V, the precision medicine technique BH3 profiling, as well as standard experimental techniques \u2013 flow cytometry, siRNA and CRISPR Cas9 genetic manipulation, Western blotting, and immunohistochemistry. These techniques were used on a comprehensive range of lymphoid malignant models including diffuse large B-cell lymphoma, Burkitt lymphoma, and T-acute lymphoblastic leukemia cell lines, patient primary chronic lymphocytic leukemia cells, as well as lymphoid cell-line derived and patient-derived xenograft mice, to elucidate the mechanism of cell death by CD47 blockade and to identify the optimal drug partners for treatment combination.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>We demonstrate that the anti-CD47 antibodies SRF231, magrolimab, and B6H12 eliminated tumor cells from various in vitro and in vivo lymphoid malignant models via the activation of the RIPK1\/MLKL\/necroptotic pathway. Moreover, the BH3 profiling technique distinguished two different lymphoid malignant models that respond differently to the BCL-2 inhibitor venetoclax when combined with SRF231; one highlighting the effective yet distinct mechanisms of SRF231-induced necroptosis and venetoclax-induced apoptosis in models that were specifically and\/or highly dependent on BCL-2 for survival, while the other implicating venetoclax as a counterproductive partner with SRF231 in models that were not dependent on BCL-2 for survival or were not responsive to venetoclax treatment.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>Collectively, this study unravels a novel, non-canonical cell death mechanism of targeting CD47 by activating necroptosis, and provides evidence and rationale for further evaluation of a therapeutic strategy of combining CD47 blockade with and without apoptotic inducers for suitable patients with lymphoid malignancies.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s13045-025-01774-3","type":"journal-article","created":{"date-parts":[[2026,1,3]],"date-time":"2026-01-03T06:45:36Z","timestamp":1767422736000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["CD47 blockade-driven necroptosis complements BCL-2 inhibition-driven apoptosis in lymphoid malignancies"],"prefix":"10.1186","volume":"19","author":[{"given":"Stephen J. 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Animal studies were approved by the Dana-Farber internal and Surface Oncology external Institutional Animal Care and Use Committees.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"R. Valentin is a current employee at Genmab. J. L. Guerriero is a consultant for Glaxo-Smith Kline (GSK), Codagenix, Verseau Therapeutics, Kymera, Kowa, Duke Street Bio., and Array BioPharma and receives sponsored research support from Duke Street Bio, GSK, Array BioPharma, Merck and Eli Lilly. T. Z. Lehmberg is a current employee at Casma Therapeutics. M. O. Peluso, A. Adam, L. Zhang, C. M. Armet, M. Rausch, B. H. Lee, V. J. Palombella, P. M. Holland, A. M. Paterson are former employees of Surface Oncology, which has been acquired by Coherus BioSciences. C. Herbaux has received honoraria and travel funding from AbbVie. C. U. Niemann has received consultancy fees and\/or travel grants from Janssen, Abbvie, Novartis, Roche, Sunesis, Gilead, AstraZeneca, and CSL Behring, research support from Abbvie, AstraZeneca, Janssen and Novo Nordisk Foundation (NNF16OC0019302), outside this project. M. S. Davids is on Consultancy\/Advisory Boards at AbbVie, Adaptive, Ascentage, Astra-Zeneca, Beigene, Eli Lilly, Galapagos, Genentech, Genmab, Janssen, MEI Pharma, Merck, Nuvalent, Schr\u04e7dinger, Takeda, and has received research funding from Ascentage, MEI Pharma, Novartis, and Surface Oncology. C. J. Wu has received research funding from Pharmacyclics. All other authors declare no competing interests.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"11"}}