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@ARTICLE{Sartorius:301917,
author = {S. Sartorius and L. Bramè$^*$ and J. Proba and A. Gauert
and N. Olk and J. Lazaro and A. Eggert and C. Eckert and A.
H. Hagemann$^*$},
title = {{R}apid in vivo {D}rug {R}esponse {P}rediction {U}sing
{L}eukemia {C}ell {G}rafts in {Z}ebrafish {E}mbryos.},
journal = {JoVE journal},
volume = {23},
number = {219},
issn = {1940-087X},
address = {Cambridge, MA},
publisher = {JoVE},
reportid = {DKFZ-2025-01187},
pages = {67451},
year = {2025},
note = {ISSN 1940-087X},
abstract = {Zebrafish xenotransplantation is a pivotal technique for
investigating human cancer pathogenesis and predicting
individual drug responses. This document introduces a
streamlined protocol (ZefiX) for expanding primary B-cell
precursor acute lymphoblastic leukemia (BCP-ALL) patient
samples or immortalized cell lines in transiently
immunosuppressed zebrafish embryos, utilizing flow cytometry
for high-resolution single-cell analysis of treatment
responses. Compared to solid tumor engraftments, leukemia
cells profit significantly from a morpholino antisense
oligonucleotide-based suppression of macrophage and
neutrophil differentiating factors during the assay. Flow
cytometry analysis of dissociated graft cells enables
precise evaluation of cell count, proliferation rate, and
vitality after treatment on a per-cell basis. This approach
has been validated using targeted therapeutics such as
venetoclax and dasatinib, with treatment outcomes compared
to clinical records of related patient samples and
traditional 2D culture controls. Notably, the protocol is
completed within 7 days, aligning with clinical
decision-making timelines. The methodology is adaptable for
testing selected drugs in various cancer types, including
solid tumors, thereby supporting personalized therapeutic
strategies. However, limitations on the number of drugs that
can be assessed, likely due to pharmacokinetic constraints
in zebrafish embryos, should be considered.},
keywords = {Zebrafish: embryology / Animals / Humans / Antineoplastic
Agents: pharmacology / Flow Cytometry: methods / Precursor
B-Cell Lymphoblastic Leukemia-Lymphoma: drug therapy /
Precursor B-Cell Lymphoblastic Leukemia-Lymphoma: pathology
/ Antineoplastic Agents (NLM Chemicals)},
cin = {BE01},
ddc = {570},
cid = {I:(DE-He78)BE01-20160331},
pnm = {899 - ohne Topic (POF4-899)},
pid = {G:(DE-HGF)POF4-899},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:40489456},
doi = {10.3791/67451},
url = {https://inrepo02.dkfz.de/record/301917},
}