دورية أكاديمية
أعرض في EDS

CAR-T cell expansion platforms yield distinct T cell differentiation states.

التفاصيل البيبلوغرافية
العنوان: CAR-T cell expansion platforms yield distinct T cell differentiation states.
المؤلفون: Song HW; Department of Transfusion Medicine, Center for Cellular Engineering, National Institutes of Health, Bethesda, MD, USA., Prochazkova M; Department of Transfusion Medicine, Center for Cellular Engineering, National Institutes of Health, Bethesda, MD, USA., Shao L; Department of Transfusion Medicine, Center for Cellular Engineering, National Institutes of Health, Bethesda, MD, USA., Traynor R; Department of Transfusion Medicine, Center for Cellular Engineering, National Institutes of Health, Bethesda, MD, USA., Underwood S; Department of Transfusion Medicine, Center for Cellular Engineering, National Institutes of Health, Bethesda, MD, USA., Black M; Department of Transfusion Medicine, Center for Cellular Engineering, National Institutes of Health, Bethesda, MD, USA., Fellowes V; Center for Immuno-Oncology, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA., Shi R; Department of Transfusion Medicine, Center for Cellular Engineering, National Institutes of Health, Bethesda, MD, USA., Pouzolles M; Pediatric Oncology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA., Chou HC; Genomics Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA., Cheuk AT; Genomics Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA., Taylor N; Pediatric Oncology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA., Jin P; Department of Transfusion Medicine, Center for Cellular Engineering, National Institutes of Health, Bethesda, MD, USA., Somerville RP; Department of Transfusion Medicine, Center for Cellular Engineering, National Institutes of Health, Bethesda, MD, USA., Stroncek DF; Department of Transfusion Medicine, Center for Cellular Engineering, National Institutes of Health, Bethesda, MD, USA., Khan J; Genomics Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA., Highfill SL; Department of Transfusion Medicine, Center for Cellular Engineering, National Institutes of Health, Bethesda, MD, USA. Electronic address: steven.highfill@nih.gov.
المصدر: Cytotherapy [Cytotherapy] 2024 Jul; Vol. 26 (7), pp. 757-768. Date of Electronic Publication: 2024 Mar 12.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: Elsevier Country of Publication: England NLM ID: 100895309 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1477-2566 (Electronic) Linking ISSN: 14653249 NLM ISO Abbreviation: Cytotherapy Subsets: MEDLINE
أسماء مطبوعة: Publication: 2013- : London : Elsevier
Original Publication: Oxford, England : ISIS Medical Media, c1999-
مواضيع طبية MeSH: Cell Differentiation* , Immunotherapy, Adoptive*/methods , Receptors, Chimeric Antigen*/genetics , Receptors, Chimeric Antigen*/metabolism, Humans ; Cell Proliferation ; T-Lymphocytes/metabolism ; T-Lymphocytes/cytology ; Bioreactors ; Cell Culture Techniques/methods ; CD8-Positive T-Lymphocytes/immunology
مستخلص: With investigators looking to expand engineered T cell therapies such as CAR-T to new tumor targets and patient populations, a variety of cell manufacturing platforms have been developed to scale manufacturing capacity using closed and/or automated systems. Such platforms are particularly useful for solid tumor targets, which typically require higher CAR-T cell doses. Although T cell phenotype and function are key attributes that often correlate with therapeutic efficacy, how manufacturing platforms influence the final CAR-T cell product is currently unknown. We compared 4 commonly used T cell manufacturing platforms (CliniMACS Prodigy, Xuri W25 rocking platform, G-Rex gas-permeable bioreactor, static bag culture) using identical media, stimulation, culture length, and donor starting material. Selected CD4 + CD8 + cells were transduced with lentiviral vector incorporating a CAR targeting FGFR4, a promising target for pediatric sarcoma. We observed significant differences in overall expansion over the 14-day culture; bag cultures had the highest capacity for expansion while the Prodigy had the lowest (481-fold versus 84-fold, respectively). Strikingly, we also observed considerable differences in the phenotype of the final product, with the Prodigy significantly enriched for CCR7 + CD45RA + naïve/stem central memory (T n/scm )-like cells at 46% compared to bag and G-Rex with 16% and 13%, respectively. Gene expression analysis also showed that Prodigy CAR-Ts are more naïve, less cytotoxic and less exhausted than bag, G-Rex, and Xuri CAR-Ts, and pointed to differences in cell metabolism that were confirmed via metabolic assays. We hypothesized that dissolved oxygen level, which decreased substantially during the final 3 days of the Prodigy culture, may contribute to the observed differences in T cell phenotype. By culturing bag and G-Rex cultures in 1% O 2 from day 5 onward, we could generate >60% T n/scm -like cells, with longer time in hypoxia correlating with a higher percentage of T n/scm -like cells. Intriguingly, our results suggest that oxygenation is responsible, at least in part, for observed differences in T cell phenotype among bioreactors and suggest hypoxic culture as a potential strategy prevent T cell differentiation during expansion. Ultimately, our study demonstrates that selection of bioreactor system may have profound effects not only on the capacity for expansion, but also on the differentiation state of the resulting CAR-T cells.
Competing Interests: Declaration of Competing Interest The authors declare no competing interests. The views expressed by the authors do not necessarily represent the views of the NIH, Department of Health and Human Services, or the US Federal Government.
(Published by Elsevier Inc.)
فهرسة مساهمة: Keywords: CAR-T cells; cell expansion; cell manufacturing; chimeric antigen receptor; hypoxia
المشرفين على المادة: 0 (Receptors, Chimeric Antigen)
تواريخ الأحداث: Date Created: 20240416 Date Completed: 20240628 Latest Revision: 20240628
رمز التحديث: 20240629
DOI: 10.1016/j.jcyt.2024.03.003
PMID: 38625071
قاعدة البيانات: MEDLINE
الوصف
تدمد:1477-2566
DOI:10.1016/j.jcyt.2024.03.003