TY  - JOUR
AU  - Sartorius, Sophia
AU  - Bramè, Luca
AU  - Proba, Jutta
AU  - Gauert, Anton
AU  - Olk, Nadine
AU  - Lazaro, Juan
AU  - Eggert, Angelika
AU  - Eckert, Cornelia
AU  - Hagemann, Anja H
TI  - Rapid in vivo Drug Response Prediction Using Leukemia Cell Grafts in Zebrafish Embryos.
JO  - JoVE journal
VL  - 23
IS  - 219
SN  - 1940-087X
CY  - Cambridge, MA
PB  - JoVE
M1  - DKFZ-2025-01187
SP  - 67451
PY  - 2025
N1  - ISSN 1940-087X
AB  - 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.
KW  - Zebrafish: embryology
KW  - Animals
KW  - Humans
KW  - Antineoplastic Agents: pharmacology
KW  - Flow Cytometry: methods
KW  - Precursor B-Cell Lymphoblastic Leukemia-Lymphoma: drug therapy
KW  - Precursor B-Cell Lymphoblastic Leukemia-Lymphoma: pathology
KW  - Antineoplastic Agents (NLM Chemicals)
LB  - PUB:(DE-HGF)16
C6  - pmid:40489456
DO  - DOI:10.3791/67451
UR  - https://inrepo02.dkfz.de/record/301917
ER  -