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Stenger, D.* ; Stief, T.A.* ; Kaeuferle, T.* ; Willier, S.* ; Rataj, F.* ; Schober, K.* ; Vick, B. ; Lotfi, R.* ; Wagner, B.* ; Gruenewald, T.G.P.* ; Kobold, S.* ; Busch, D.H.* ; Jeremias, I. ; Blaeschke, F.* ; Feuchtinger, T.*

Endogenous TCR promotes in vivo persistence of CD19-CAR-T cells compared to a CRISPR/Cas9-mediated TCR knockout CAR.

Blood 136, 1407-1418 (2020)
: Publ. Version/Full Text online available 10/2021
Anti-CD19 chimeric antigen receptor (CAR) T cells showed significant antileukemic activity in B-precursor acute lymphoblastic leukemia (ALL). Allogeneic, HLA-mismatched off-the-shelf third-party donors may offer ideal fitness of the effector cells, but carry the risk of graft-versus-host disease. Knockout (KO) of the endogenous T-cell receptor (TCR) in CD19-CAR-T cells may be a promising solution. Here, we induced a CRISPR/Cas9-mediated KO of the TCR chain in combination with a second-generation retroviral CAR transduction including a 4-1BB costimulatory domain in primary T cells. This tandem engineering led to a highly functional population of TCR-KO-CAR-T cells with strong activation (CD25, interferon gamma), proliferation, and specific killing upon CD19 target recognition. TCR-KO-CART cells had a balanced phenotype of central memory and effector memory T cells. KO of the endogenous TCR in T cells strongly ablated alloreactivity in comparison with TCR-expressing T cells. In a patient-derived xenograft model of childhood ALL TCR-KO-CAR-T cells clearly controlled CD19(+) leukemia burden and improved survival in vivo. However, coexpression of endogenous TCR plus CAR led to superior persistence of T cells and significantly prolonged leukemia control in vivo, confirmed by a second in vivo model using the leukemia cell line NALM6. These results point toward an essential role of the endogenous TCR for longevity of the response at the price of alloreactivity. In conclusion, anti-CD19 CAR T cells with a CRISPR/Cas9-mediated TCR-KO are promising candidates for nonmatched thirdparty adoptive T-cell transfer with high antileukemic functionality in the absence of alloreactivity, but long-term persistence in vivo is better in the presence of the endogenous TCR.
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Publication type Article: Journal article
Document type Scientific Article
Keywords T-cells; Antitumor Immunity; Children; Leukemia; Immunotherapy; Expression; Receptor; Subset; Il-15
ISSN (print) / ISBN 0006-4971
e-ISSN 1528-0020
Journal Blood
Quellenangaben Volume: 136, Issue: 12, Pages: 1407-1418 Article Number: , Supplement: ,
Publisher American Society of Hematology
Publishing Place 2021 L St Nw, Suite 900, Washington, Dc 20036 Usa
Reviewing status Peer reviewed
Institute(s) Research Unit Apoptosis in Hematopoietic Stem Cells (AHS)
Grants Gert and Susanna Mayer foundation
Bettina Brau Stiftung
Gesellschaft fur Kinderkrebsforschung
Dr. Sepp und Hanne Sturm Gedaechtnisstiftung
Gertrud und Hugo Adler Stiftung
Gottfried Kieser-Stiftung
Else-Kroner-Fresenius Stiftung
German Cancer Research Center/German Cancer Consortium
European Research Council
ERC Consolidator Grant
Mildred Scheel Professorship from German Cancer Aid
German Cancer Aid
Kinderkrebshilfe Ebersberg e.V.