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@ARTICLE{Teibo:303211,
author = {J. O. Teibo and R. M. Silveira and V. C. Silvestrini and I.
Archiolli and A. PaulaMasson and B. P. de Morais and D.
Schmidt and M. H. Dos Santos and G. A. Ferreira and C. H.
Thomé and D. Helm$^*$ and R. S. Nirujogi and D. R. Alessi
and V. Picanço-Castro and L. E. B. de Souza and V. M.
Faça},
title = {{P}roteomics analysis reveals early event molecular
effectors of anti-{CD}19 {CAR}-{T} cell therapy in
hematological cancer.},
journal = {Journal of proteomics},
volume = {321},
issn = {1874-3919},
address = {New York, NY [u.a.]},
publisher = {Elsevier},
reportid = {DKFZ-2025-01562},
pages = {105507},
year = {2025},
note = {Volume 321, 30 October 2025, 105507Journal of Proteomics},
abstract = {Chimeric antigen receptor T-cell (CAR-T) therapy is at the
forefront of the field of cell immunotherapy. In this study,
we generated an anti-CD19 CAR-Jurkat T cell line using a
locally produced second-generation anti-CD19 CAR construct,
which allowed us to analyse early proteomic changes that are
crucial for comprehending the signalling pathways and
mechanism of action of this CAR-T cell. SILAC-heavy tagged
RAJI B-cells and anti-CD19 CAR-Jurkat T-cells were
co-cultured for ten minutes. The proteomic profiles were
acquired via DIA methodology on the Orbitrap Astral LC-MS/MS
platform. The proteome was extensively covered, resulting in
about 8800 protein identifications at 1 $\%$ FDR. The
effector CAR-Jurkat cells showed proteomic changes involving
antigen presentation by CD74. The target RAJI B-cells
exhibited more significant alterations. Effector proteins,
namely CD247, CD28, DAP, LCK, p38 MAPK, and CASP3, were
validated, as they have critical roles in antigen
presentation, T-cell activation, and apoptosis.
Pharmacological inhibition of LCK using Dasatinib further
suggested its pivotal role in early CAR-T signalling. This
study led us to identify proteins that function as molecular
effectors of anti-CD19 CAR-T cell therapy during the initial
phases of CAR-T-target cell engagement, advancing our
knowledge of the mechanism and signalling pathways that will
support CAR-T cell development. SIGNIFICANCE: Chimeric
antigen receptor T-cell (CAR-T cell) therapy is
state-of-the-art in cell and immunotherapy. Determining
important players in cellular communication and signalling
mediated by membranes and intracellular proteins require
understanding the connection between tumours and modified
cells. We employed global proteomics in this study to better
grasp the functional protein networks using a
high-sensitivity mass spectrometric platform for protein
identification and quantification. We identified proteins as
molecular effectors of anti-CD19 CAR-T cell treatment during
the early stages of CAR-T-target cell interaction. Our
understanding of the mechanism and signalling pathways will
promote the development of new CAR constructs and improve
the efficacy and ability to overcome the resistance of this
innovative cancer treatment strategy, which will advance the
identification of adjuvant molecules for the regulation of
CAR-T responses.},
keywords = {CAR-T cell (Other) / Early Signalling event (Other) /
Hematological Cancer (Other) / Molecular effectors (Other) /
Proteomics (Other) / SILAC (Other)},
cin = {W120},
ddc = {540},
cid = {I:(DE-He78)W120-20160331},
pnm = {312 - Funktionelle und strukturelle Genomforschung
(POF4-312)},
pid = {G:(DE-HGF)POF4-312},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:40716489},
doi = {10.1016/j.jprot.2025.105507},
url = {https://inrepo02.dkfz.de/record/303211},
}