Targeted therapy of human leukemia xenografts in immunodeficient zebrafish

Abstract

Personalized medicine holds tremendous promise for improving safety and efficacy of drug therapies by optimizing treatment regimens. Rapidly developed patient-derived xenografts (pdx) could be a helpful tool for analyzing the effect of drugs against an individual’s tumor by growing the tumor in an immunodeficient animal. Severe combined immunodeficiency (SCID) mice enable efficient in vivo expansion of vital tumor cells and generation of personalized xenografts. However, they are not amenable to large-scale rapid screening, which is critical in identifying new compounds from large compound libraries. The development of a zebrafish model suitable for pdx could facilitate large-scale screening of drugs targeted against specific malignancies. Here, we describe a novel strategy for establishing a zebrafish model for drug testing in leukemia xenografts. We used chronic myelogenous leukemia and acute myeloid leukemia for xenotransplantation into SCID zebrafish to evaluate drug screening protocols. We showed the in vivo efficacy of the ABL inhibitor imatinib, MEK inhibitor U0126, cytarabine, azacitidine and arsenic trioxide. We performed corresponding in vitro studies, demonstrating that combination of MEK- and FLT3-inhibitors exhibit an enhanced effect in vitro. We further evaluated the feasibility of zebrafish for transplantation of primary human hematopoietic cells that can survive at 15 day-post-fertilization. Our results provide critical insights to guide development of high-throughput platforms for evaluating leukemia.

Bioluminescent Zebrafish Transplantation Model for Drug Discovery

“…Grafted embryos and larvae could be treated by simply soaking them in chemical compounds dissolved in water. Cell proliferation is most often measured until 3–4 dpi by the quantification of fluorescently labeled cancer cells, in vivo ( Corkery et al, 2011 ; Ghotra et al, 2012 ; Tulotta et al, 2016 ; Hason and Bartunek, 2019 ; Tulotta et al, 2019 ; Patton et al, 2021b ) or ex vivo ( Somasagara et al, 2021 ). Our screening workflow enables rapid characterization of tumor-inhibitor responsiveness within a time frame of 7 days.…”

“…Targeting of cancer-related pathways in zebrafish embryos assisted in the discovery of compounds such as leflunomide ( White et al, 2011 ), Lenaldekar ( Ridges et al, 2012 ), perphenazine ( Gutierrez et al, 2014 ), and clotrimazole ( Precazzini et al, 2020 ) as potential treatment options for cancer mono- and combined therapy of human leukemia and melanoma. Even though there are many established zebrafish xenograft models, up until now, there were only a few larger inhibitor screens done in zebrafish allograft or xenograft models ( Tulotta et al, 2016 ; Wertman et al, 2016 ; Fior et al, 2017 ; Fazio et al, 2020 ; Almstedt et al, 2021 ; Somasagara et al, 2021 ). This is likely due to the limited options of workflow automation in transplantation studies.…”

Bioluminescent zebrafish transplantation model for drug discovery

“…Grafted embryos and larvae could be treated by simply soaking them in chemical compounds dissolved in water. Cell proliferation is most often measured until 3-4 dpi by the quantification of fluorescently labeled cancer cells, in vivo (Corkery et al, 2011; Ghotra et al, 2012; Hason and Bartunek, 2019; Patton et al, 2021b; Tulotta et al, 2016; Tulotta et al, 2019) or ex vivo (Somasagara et al, 2021). Our screening workflow enables rapid characterization of tumor-inhibitor responsiveness within a time frame of seven days.…”

“…Targeting of cancer-related pathways in zebrafish embryos assisted in the discovery of compounds such as leflunomide (White et al, 2011), Lenaldekar (Ridges et al, 2012), perphenazine (Gutierrez et al, 2014), and clotrimazole (Precazzini et al, 2020) as potential treatment options for cancer mono- and combined therapy of human leukemia and melanoma. Even though there are many established zebrafish xenograft models, up until now, there were only a few larger inhibitor screens done in zebrafish allograft or xenograft models (Almstedt et al, 2021; Fazio et al, 2020; Fior et al, 2017; Somasagara et al, 2021; Tulotta et al, 2016; Wertman et al, 2016). This is likely due to the limited options of workflow automation in transplantation studies.…”

One Host-Multiple Applications: Zebrafish (Danio rerio) as Promising Model for Studying Human Cancers and Pathogenic Diseases

“…The results of histology, RNA sequencing, and single-cell transcriptional profiling showed a decrease in mature T and B cells in prkdc -deficient ZF [ 122 ]. This model has been successfully used to engraft human leukemia cells and drug tests [ 123 ]. Another model based on prkdc −/−, il2rga −/− zebrafish has been used as PDX for melanoma, breast cancer embryonal rhabdomyosarcoma (ERMS), and glioblastoma.…”