{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,3]],"date-time":"2026-11-03T21:39:52Z","timestamp":1793741992089,"version":"4.6.1"},"reference-count":53,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,2,9]],"date-time":"2022-02-09T00:00:00Z","timestamp":1644364800000},"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. Immunol."],"abstract":"<jats:p>CAR T-cell therapies targeting the B-cell maturation antigen eliminate tumors in relapsed\/refractory multiple myeloma patients, however durable remissions remain difficult to attain. Transforming growth factor beta (TGF-\u03b2) is a multifunctional cytokine abundantly expressed in the multiple myeloma bone marrow niche, where it promotes an immunosuppressive tumor microenvironment. We hypothesized that BCMA CAR T-cells armored to resist the suppressive effects of TGF-\u03b2 will provide an advantage in treating multiple myeloma. The armored B2ARM CAR T cells, co-expressing BCMA targeting CAR with TGF-\u03b2 dominant-negative receptor II, were generated by lentiviral transduction of primary human CD4+ and CD8+ T cells. The B2ARM CAR T cells eliminated MM.1S multiple myeloma targets in long-term cytotoxicity assays, even under TGF-\u03b2-high conditions, whereas cytotoxic function of the non-armored B2 CAR -T cells was inhibited by TGF-\u03b2. Concordantly, after long-term exposure to targets in the presence of TGF-\u03b2, the B2ARM CAR T cells were enriched for Granzyme B, CD107a, Ki67 and polyfunctional cells T-cells (double or triple-positive for IFN-\u03b3, IL-2 and\/or TNF-\u03b1), as determined by flow cytometry. In addition, the B2ARM CAR T-cells, but not the conventional B2 CAR T-cells, resisted the TGF-\u03b2-mediated suppression of activation (CD25), exhaustion (PD-1, LAG3), and differentiation to T effectors (CD45RA+ CD45RO-CD62L-). In NSG mice bearing RPMI-8226 tumors overexpressing TGF-\u03b2, the B2ARM CAR mediated 100% tumor rejection and survival, superior infiltration of tumors on day 7 post CAR T treatment (%CD3+CAR+), and greater expression of IFN-\u03b3, TNF-\u03b1, Ki67, Granzyme B, and PD-1, as compared to tumor-infiltrating non-armored B2 CAR T-cells. In NSG RPMI-8226 xenograft model in which tumors were additionally supplemented with TGF-\u03b2 injections on days -1 through 11 of CAR T treatment, the B2ARM CAR T cells rejected tumors faster than the non-armored B2 CARs, and showed greater numbers of CD3+ and CD3+CAR+, central memory (CD45RO+CD62L+) and effector memory (CD45RO+CD62L-) T cells in the peripheral blood 18 days after treatment. In summary, the armored B2ARM CAR T cells mediate superior persistence, proliferation, multi-functionality, effector differentiation and anti-tumor function in pre-clinical models of multiple myeloma, while abrogating TGF-\u03b2-mediated suppression.<\/jats:p>","DOI":"10.3389\/fimmu.2022.832645","type":"journal-article","created":{"date-parts":[[2022,2,8]],"date-time":"2022-02-08T23:31:46Z","timestamp":1644363106000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":60,"title":["Armored BCMA CAR T Cells Eliminate Multiple Myeloma and Are Resistant to the Suppressive Effects of TGF-\u03b2"],"prefix":"10.3389","volume":"13","author":[{"given":"Leah M.","family":"Alabanza","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying","family":"Xiong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bang","family":"Vu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brian","family":"Webster","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Darong","family":"Wu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peirong","family":"Hu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhongyu","family":"Zhu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Boro","family":"Dropulic","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pradyot","family":"Dash","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dina","family":"Schneider","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2022,2,9]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"125","DOI":"10.1186\/s13045-020-00962-7","article-title":"BCMA-Targeted Immunotherapy for Multiple Myeloma","volume":"13","author":"Yu","year":"2020","journal-title":"J Hematol Oncol"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1111\/ejh.13380","article-title":"B Cell Maturation Antigen-Specific Chimeric Antigen Receptor T Cells for Relapsed or Refractory Multiple Myeloma: A Meta-Analysis","volume":"104","author":"Gagelmann","year":"2020","journal-title":"Eur J Haematol"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1080\/10428194.2020.1827253","article-title":"Trends in the Multiple Myeloma Treatment Landscape and Survival: A U.S. Analysis Using 2011-2019 Oncology Clinic Electronic Health Record Data","author":"Braunlin","year":"","journal-title":"Leukemia Lymphoma"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2020.01128","article-title":"Overcoming Chimeric Antigen Receptor (CAR) Modified T-Cell Therapy Limitations in Multiple Myeloma","volume":"11","author":"Garcia-Guerrero","year":"2020","journal-title":"Front Immunol"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1200\/JCO.2018.77.8084","article-title":"T Cells Genetically Modified to Express an Anti-B-Cell Maturation Antigen Chimeric Antigen Receptor Cause Remissions of Poor-Prognosis Relapsed Multiple Myeloma","volume":"36","author":"Brudno","year":"2018","journal-title":"J Clin Oncol"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1172\/JCI126397","article-title":"B Cell Maturation Antigen-Specific CAR T Cells Are Clinically Active in Multiple Myeloma","volume":"129","author":"Cohen","year":"2019","journal-title":"J Clin Invest"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2016-04-711903","article-title":"T Cells Expressing an Anti-B-Cell Maturation Antigen Chimeric Antigen Receptor Cause Remissions of Multiple Myeloma","volume":"128","author":"Ali","year":"2016","journal-title":"Blood"},{"key":"B8","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1186\/s13045-018-0681-6","article-title":"A Phase 1, Open-Label Study of LCAR-B38M, a Chimeric Antigen Receptor T Cell Therapy Directed Against B Cell Maturation Antigen, in Patients With Relapsed or Refractory Multiple Myeloma","volume":"11","author":"Zhao","year":"2018","journal-title":"J Hematol Oncol"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1817226","article-title":"Anti-BCMA CAR T-Cell Therapy Bb2121 in Relapsed or Refractory Multiple Myeloma","volume":"380","author":"Raje","year":"2019","journal-title":"New Engl J Med"},{"key":"B10","doi-asserted-by":"publisher","first-page":"157496","DOI":"10.1155\/2012\/157496","article-title":"Bone Marrow Microenvironment in Multiple Myeloma Progression","volume":"2012","author":"Manier","year":"2012","journal-title":"J Biomed Biotechnol"},{"key":"B11","doi-asserted-by":"publisher","first-page":"21","DOI":"10.1007\/s00109-015-1345-4","article-title":"Dissecting the Multiple Myeloma-Bone Microenvironment Reveals New Therapeutic Opportunities","volume":"94","author":"Shay","year":"2016","journal-title":"J Mol Med"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1196\/annals.1346.035","article-title":"Bone Destruction in Multiple Myeloma","volume":"1068","author":"Matsumoto","year":"2006","journal-title":"Ann New York Acad Sci"},{"key":"B13","doi-asserted-by":"publisher","first-page":"3099","DOI":"10.3390\/cancers12113099","article-title":"Transforming Growth Factor-Beta: A Multifunctional Regulator of Cancer Immunity","volume":"12","author":"Xue","year":"2020","journal-title":"Cancers"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2005-10-4169","article-title":"Role of Transforming Growth Factor-Beta in Hematologic Malignancies","volume":"107","author":"Dong","year":"2006","journal-title":"Blood"},{"key":"B15","doi-asserted-by":"publisher","first-page":"e9870","DOI":"10.1371\/journal.pone.0009870","article-title":"Tgf-Beta Inhibition Restores Terminal Osteoblast Differentiation to Suppress Myeloma Growth","volume":"5","author":"Takeuchi","year":"2010","journal-title":"PloS One"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V87.5.1928.1928","article-title":"Transforming Growth Factor-Beta1: Differential Effects on Multiple Myeloma Versus Normal B Cells","volume":"87","author":"Urashima","year":"1996","journal-title":"Blood"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1016\/j.bone.2010.05.036","article-title":"TGF-Beta-Related Mechanisms of Bone Destruction in Multiple Myeloma","volume":"48","author":"Matsumoto","year":"2011","journal-title":"Bone"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1016\/0006-291X(89)92795-2","article-title":"Activation of the Bone-Derived Latent TGF Beta Complex by Isolated Osteoclasts","volume":"158","author":"Oreffo","year":"1989","journal-title":"Biochem Biophys Res Commun"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1002\/jbmr.5650090402","article-title":"Osteoclast Synthesis and Secretion and Activation of Latent Transforming Growth Factor Beta","volume":"9","author":"Oursler","year":"1994","journal-title":"J Bone Mineral Res"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1038\/sj.emboj.7600067","article-title":"Endogenous TGF-Beta Signaling Suppresses Maturation of Osteoblastic Mesenchymal Cells","volume":"23","author":"Maeda","year":"2004","journal-title":"EMBO J"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1093\/emboj\/20.9.2254","article-title":"TGF-Beta-Induced Repression of CBFA1 by Smad3 Decreases Cbfa1 and Osteocalcin Expression and Inhibits Osteoblast Differentiation","volume":"20","author":"Alliston","year":"2001","journal-title":"EMBO J"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2009-10-246660","article-title":"Elevated IL-17 Produced by TH17 Cells Promotes Myeloma Cell Growth and Inhibits Immune Function in Multiple Myeloma","volume":"115","author":"Prabhala","year":"2010","journal-title":"Blood"},{"key":"B23","doi-asserted-by":"publisher","first-page":"392","DOI":"10.1016\/j.cell.2008.07.025","article-title":"TGF-Beta: A Master of All T Cell Trades","volume":"134","author":"Li","year":"2008","journal-title":"Cell"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.173.5.3093","article-title":"p21Cip1 and p27Kip1 Act in Synergy to Alter the Sensitivity of Naive T Cells to TGF-Beta-Mediated G1 Arrest Through Modulation of IL-2 Responsiveness","volume":"173","author":"Wolfraim","year":"2004","journal-title":"J Immunol"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1084\/jem.189.2.231","article-title":"Transforming Growth Factor Beta1 Inhibits Fas Ligand Expression and Subsequent Activation-Induced Cell Death in T Cells via Downregulation of C-Myc","volume":"189","author":"Genestier","year":"1999","journal-title":"J Exp Med"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.172.7.4275","article-title":"Smad3 is Essential for TGF-Beta 1 to Suppress IL-2 Production and TCR-Induced Proliferation, But Not IL-2-Induced Proliferation","volume":"172","author":"McKarns","year":"2004","journal-title":"J Immunol"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1002\/jlb.66.6.981","article-title":"Transforming Growth Factor Beta From Multiple Myeloma Cells Inhibits Proliferation and IL-2 Responsiveness in T Lymphocytes","volume":"66","author":"Cook","year":"1999","journal-title":"J Leukocyte Biol"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1016\/j.ccr.2005.10.012","article-title":"TGF-Beta Directly Targets Cytotoxic T Cell Functions During Tumor Evasion of Immune Surveillance","volume":"8","author":"Thomas","year":"2005","journal-title":"Cancer Cell"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20012076","article-title":"Mechanism of Transforming Growth Factor Beta-Induced Inhibition of T Helper Type 1 Differentiation","volume":"195","author":"Gorelik","year":"2002","journal-title":"J Exp Med"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.174.10.5950","article-title":"TGF-Beta 1 Uses Distinct Mechanisms to Inhibit IFN-Gamma Expression in CD4+ T Cells at Priming and at Recall: Differential Involvement of Stat4 and T-Bet","volume":"174","author":"Lin","year":"2005","journal-title":"J Immunol"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.165.9.4773","article-title":"Cutting Edge: TGF-Beta Inhibits Th Type 2 Development Through Inhibition of GATA-3 Expression","volume":"165","author":"Gorelik","year":"2000","journal-title":"J Immunol"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1038\/nature04753","article-title":"Reciprocal Developmental Pathways for the Generation of Pathogenic Effector TH17 and Regulatory T Cells","volume":"441","author":"Bettelli","year":"2006","journal-title":"Nature"},{"key":"B33","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1093\/jmcb\/mjr033","article-title":"TGF-Beta: The Sword, the Wand, and the Shield of FOXP3(+) Regulatory T Cells","volume":"4","author":"Tran","year":"2012","journal-title":"J Mol Cell Biol"},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.1002\/hon.2206","article-title":"Upregulation of CD200 is Associated With Regulatory T Cell Expansion and Disease Progression in Multiple Myeloma","volume":"35","author":"Aref","year":"2017","journal-title":"Hematol Oncol"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1159\/000203602","article-title":"Overproduction of IL-7, IL-10 and TGF-Beta 1 in Multiple Myeloma","volume":"98","author":"Kroning","year":"1997","journal-title":"Acta Haematol"},{"key":"B36","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1159\/000203962","article-title":"Increased Intraplatelet and Urinary Transforming Growth Factor-Beta in Patients With Multiple Myeloma","volume":"94","author":"Jiang","year":"1995","journal-title":"Acta Haematol"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1177\/030089160409000404","article-title":"Effects of Interferon-Alpha-2a on Th3 Cytokine Response in Multiple Myeloma Patients","volume":"90","author":"Sonmez","year":"2004","journal-title":"Tumori"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2249.1990.tb05134.x","article-title":"Defective Interleukin-2 Induction of Lymphokine-Activated Killer (LAK) Activity in Peripheral Blood T Lymphocytes of Patients With Monoclonal Gammopathies","volume":"79","author":"Massaia","year":"1990","journal-title":"Clin Exp Immunol"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V85.12.3679.bloodjournal85123679","article-title":"Dysregulated Fas and Bcl-2 Expression Leading to Enhanced Apoptosis in T Cells of Multiple Myeloma Patients","volume":"85","author":"Massaia","year":"1995","journal-title":"Blood"},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.7326\/0003-4819-41-6-1152","article-title":"Recurrent Pneumonia in Multiple Myeloma and Some Observations on Immunologic Response","volume":"41","author":"Zinneman","year":"1954","journal-title":"Ann Internal Med"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V98.10.2992","article-title":"Dendritic Cells From Patients With Myeloma Are Numerically Normal But Functionally Defective as They Fail to Up-Regulate CD80 (B7-1) Expression After Hucd40lt Stimulation Because of Inhibition by Transforming Growth Factor-Beta1 and Interleukin-10","volume":"98","author":"Brown","year":"2001","journal-title":"Blood"},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.18632\/oncotarget.17538","article-title":"MM-BMSCs Induce Naive CD4+ T Lymphocytes Dysfunction Through Fibroblast Activation Protein Alpha","volume":"8","author":"Wu","year":"2017","journal-title":"Oncotarget"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.1128\/mcb.13.12.7239-7247.1993","article-title":"Signaling Activity of Transforming Growth Factor Beta Type II Receptors Lacking Specific Domains in the Cytoplasmic Region","volume":"13","author":"Wieser","year":"1993","journal-title":"Mol Cell Biol"},{"key":"B44","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymthe.2021.05.006","article-title":"Self-Driving Armored CAR-T Cells Overcome a Suppressive Milieu and Eradicate CD19+ Raji Lymphoma in Preclinical Models","volume":"29","author":"Webster","year":"2021","journal-title":"Mol Ther"},{"key":"B45","doi-asserted-by":"publisher","first-page":"99","DOI":"10.1146\/annurev.immunol.24.021605.090737","article-title":"Transforming Growth Factor-Beta Regulation of Immune Responses","volume":"24","author":"Li","year":"2006","journal-title":"Annu Rev Immunol"},{"key":"B46","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.a.21015","article-title":"SPICE: Exploration and Analysis of Post-Cytometric Complex Multivariate Datasets","volume":"2","author":"Roederer","year":"2011","journal-title":"Cytometry"},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.3389\/fonc.2020.01117","article-title":"Arming Anti-EGFRvIII CAR-T With TGFbeta Trap Improves Antitumor Efficacy in Glioma Mouse Models","volume":"10","author":"Li","year":"2020","journal-title":"Front Oncol"},{"key":"B48","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymthe.2018.05.003","article-title":"Dominant-Negative TGF-Beta Receptor Enhances PSMA-Targeted Human CAR T Cell Proliferation And Augments Prostate Cancer Eradication","volume":"26","author":"Kloss","year":"2018","journal-title":"Mol Ther"},{"key":"B49","doi-asserted-by":"publisher","DOI":"10.1200\/JCO.2017.74.3179","article-title":"Tumor-Specific T-Cells Engineered to Overcome Tumor Immune Evasion Induce Clinical Responses in Patients With Relapsed Hodgkin Lymphoma","volume":"36","author":"Bollard","year":"2018","journal-title":"J Clin Oncol"},{"key":"B50","doi-asserted-by":"publisher","first-page":"765","DOI":"10.1053\/j.gastro.2014.07.018","article-title":"Cell Death and Cell Death Responses in Liver Disease: Mechanisms and Clinical Relevance","volume":"147","author":"Luedde","year":"2014","journal-title":"Gastroenterology"},{"key":"B51","doi-asserted-by":"publisher","DOI":"10.2217\/hep.15.27","article-title":"Apoptosis in Liver Carcinogenesis and Chemotherapy","volume":"2","author":"Moreno-Caceres","year":"2015","journal-title":"Hepatic Oncol"},{"key":"B52","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2018-01-828343","article-title":"Preinfusion Polyfunctional Anti-CD19 Chimeric Antigen Receptor T Cells Are Associated With Clinical Outcomes in NHL","volume":"132","author":"Rossi","year":"2018","journal-title":"Blood"},{"key":"B53","doi-asserted-by":"publisher","DOI":"10.1002\/jbmr.3859","article-title":"TGFbeta Inhibition Stimulates Collagen Maturation to Enhance Bone Repair and Fracture Resistance in a Murine Myeloma Model","volume":"34","author":"Green","year":"2019","journal-title":"J Bone Mineral Res"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2022.832645\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,2,8]],"date-time":"2022-02-08T23:31:52Z","timestamp":1644363112000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2022.832645\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,2,9]]},"references-count":53,"alternative-id":["10.3389\/fimmu.2022.832645"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2022.832645","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,2,9]]},"article-number":"832645"}}