Quantitative phenotyping-based in vivo chemical screening in a zebrafish model of leukemia stem cell xenotransplantation.
Zebrafish-based chemical screening has recently emerged as a rapid and efficient method to identify important compounds that modulate specific biological processes and to test the therapeutic efficacy in disease models, including cancer. In leukemia, the ablation of leukemia stem cells (LSCs) is necessary to permanently eradicate the leukemia cell population. However, because of the very small number of LSCs in leukemia cell populations, their use in xenotransplantation studies (in vivo) and the difficulties in functionally and pathophysiologically replicating clinical conditions in cell culture experiments (in vitro), the progress of drug discovery for LSC inhibitors has been painfully slow. In this study, we developed a novel phenotype-based in vivo screening method using LSCs xenotransplanted into zebrafish. Aldehyde dehydrogenase-positive (ALDH+) cells were purified from chronic myelogenous leukemia K562 cells tagged with a fluorescent protein (Kusabira-orange) and then implanted in young zebrafish at 48 hours post-fertilization. Twenty-four hours after transplantation, the animals were treated with one of eight different therapeutic agents (imatinib, dasatinib, parthenolide, TDZD-8, arsenic trioxide, niclosamide, salinomycin, and thioridazine). Cancer cell proliferation, and cell migration were determined by high-content imaging. Of the eight compounds that were tested, all except imatinib and dasatinib selectively inhibited ALDH+ cell proliferation in zebrafish. In addition, these anti-LSC agents suppressed tumor cell migration in LSC-xenotransplants. Our approach offers a simple, rapid, and reliable in vivo screening system that facilitates the phenotype-driven discovery of drugs effective in suppressing LSCs.
Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics, Mie University Graduate School of Medicine, Edobashi, Tsu, Mie, Japan.
Beibei
B
Zhang
Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics, Mie University Graduate School of Medicine, Edobashi, Tsu, Mie, Japan ; Mie University Medical Zebrafish Research Center, Edobashi, Tsu, Mie, Japan ; Department of Bioinformatics, Mie University Life Science Research Center, Edobashi, Tsu, Mie, Japan ; Department of Omics Medicine, Mie University Industrial Technology Innovation, Edobashi, Tsu, Mie, Japan ; Department of Systems Pharmacology, Mie University Graduate School of Medicine, Edobashi, Tsu, Mie, Japan.
Yasuhito
Y
Shimada
Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics, Mie University Graduate School of Medicine, Edobashi, Tsu, Mie, Japan.
Junya
J
Kuroyanagi
Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics, Mie University Graduate School of Medicine, Edobashi, Tsu, Mie, Japan ; Department of Systems Pharmacology, Mie University Graduate School of Medicine, Edobashi, Tsu, Mie, Japan.
Noriko
N
Umemoto
Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics, Mie University Graduate School of Medicine, Edobashi, Tsu, Mie, Japan ; Mie University Medical Zebrafish Research Center, Edobashi, Tsu, Mie, Japan ; Department of Bioinformatics, Mie University Life Science Research Center, Edobashi, Tsu, Mie, Japan ; Department of Omics Medicine, Mie University Industrial Technology Innovation, Edobashi, Tsu, Mie, Japan ; Department of Systems Pharmacology, Mie University Graduate School of Medicine, Edobashi, Tsu, Mie, Japan.
Yuhei
Y
Nishimura
Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharmacoinformatics, Mie University Graduate School of Medicine, Edobashi, Tsu, Mie, Japan ; Mie University Medical Zebrafish Research Center, Edobashi, Tsu, Mie, Japan ; Department of Bioinformatics, Mie University Life Science Research Center, Edobashi, Tsu, Mie, Japan ; Department of Omics Medicine, Mie University Industrial Technology Innovation, Edobashi, Tsu, Mie, Japan ; Department of Systems Pharmacology, Mie University Graduate School of Medicine, Edobashi, Tsu, Mie, Japan.
Toshio
T
Tanaka
D000444:Aldehyde Dehydrogenase / Q000378:metabolism; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D001483:Base Sequence; D017931:DNA Primers; D006801:Humans; D020014:K562 Cells; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000628:therapy*; D014411:Neoplastic Stem Cells / Q000187:drug effects / Q000637:transplantation*; D010641:Phenotype; D016133:Polymerase Chain Reaction; D033581:Stem Cell Transplantation; D014183:Transplantation, Heterologous; D015027:Zebrafish
D000970:Antineoplastic Agents; D017931:DNA Primers; D000444:Aldehyde Dehydrogenase