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@ARTICLE{Kaeuferle:300186,
author = {T. Kaeuferle and M. Zwermann and N. Stoll and P.
Ferrada-Ernst and L. Jablonowski and R. Zeidler and S.
Willier and D. Stenger and A. Yassin and R. Stripecke and T.
Feuchtinger$^*$},
title = {{A}ll-in-one {CRISPR}/{C}as-engineered
glucocorticoid-receptor knock-out {EBV}-gp350-{CAR} knock-in
{T} cells are potent and resistant to dexamethasone.},
journal = {Experimental hematology $\&$ oncology},
volume = {14},
number = {1},
issn = {2162-3619},
address = {London},
publisher = {Biomed Central},
reportid = {DKFZ-2025-00663},
pages = {40},
year = {2025},
abstract = {Epstein-Barr virus (EBV) reactivation in immunocompromised
patients and post-transplantation is associated with
morbidity, mortality and with the onset of a variety of
malignant diseases. Adoptive T-cell therapies have emerged
as promising therapeutic options, but post-transplant
immunosuppression jeopardizes the protective anti-EBV immune
surveillance by adoptively transferred T cells.Using an
all-in-one CRISPR/Cas-mediated approach, we inserted an
anti-EBV (gp350) CAR into the T-cell receptor (TRAC) locus
and simultaneously knocked-out the glucocorticoid receptor
(GR) on a good manufacturing practice (GMP)-compatible
platform.CAR knock-in (CARKI) was confirmed in primary human
T cells on genetic and on protein level with a mean
efficiency of $41\%.$ With $83\%,$ additional GR knock-out
was highly efficient in CARKI cells. On a functional level
CARKIGRKO T cells showed target-specific potency in terms of
cytokine secretion patterns, proliferative capacity and
cytotoxic activity against gp350-expressing target cells.
Further, CARKIGRKO T cells were insensitive to dexamethasone
treatment and maintained T-cell functionality. In contrast,
CARKIGRKO T cells were sensitive to the GR-independent
immunosuppressant cyclosporine A (CsA), thereby providing a
rescue treatment for patients in case of safety issues.The
study lays the proof-of-concept for virus-free all-in-one
GMP-manufacturing of glucocorticoid-resistant CAR T-cell
products. Further, the glucocorticoid-resistant gp350-CAR T
cells can provide a future therapeutic option for high-risk
post-transplant patients with EBV-reactivations or patients
with EBV-associated pathologies requiring steroid
treatment.},
keywords = {Adoptive T cell transfer (Other) / CAR T cells (Other) /
CRISPR/Cas (Other) / Dexamethasone (Other) / EBV (Other) /
Genetic engineering (Other) / Glucocorticoid receptor
(Other) / Steroid treatment (Other)},
cin = {FR01},
ddc = {610},
cid = {I:(DE-He78)FR01-20160331},
pnm = {899 - ohne Topic (POF4-899)},
pid = {G:(DE-HGF)POF4-899},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:40108708},
pmc = {pmc:PMC11921674},
doi = {10.1186/s40164-025-00631-w},
url = {https://inrepo02.dkfz.de/record/300186},
}