Transplantation of Zebrafish Pediatric Brain Tumors into Immune-competent Hosts for Long-term Study of Tumor Cell Behavior and Drug Response

Casey, Mattie J.; Modzelewska, Katarzyna; Anderson, Daniela; Goodman, James R.; Boer, Elena F.; Jiménez, Laura; Grossman, Douglas; Stewart, Rodney A.

doi:10.3791/55712-v

Cited by 2 publications

(3 citation statements)

References 23 publications

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“…Cell suspensions were maintained at 28°C during the transplantation procedure. 2dpf CG1 strain syngeneic zebrafish larvae were injected with 1-2nL of cell suspension into the hindbrain ventricle, as previously described (Casey et al, 2017). Cells were injected alone or in combination with 1% total volume Clodrosomes (Encapsula Nano Sciences, CLD-8901), vehicle control liposomes, or 0.05ng/nL human CSF-1 recombinant protein (Cell Signaling Technology, 8929SC).…”

Section: Methodsmentioning

confidence: 99%

A syngeneic spontaneous zebrafish model oftp53-deficient, EGFR^vIII, and PI3KCA^H1047R-driven glioblastoma reveals inhibitory roles for inflammation during tumor initiation and relapsein vivo

Weiss,

D’Amata,

Pearson

et al. 2023

Preprint

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To build a patient-relevant in vivo model of human glioblastoma, we expressed common oncogenic variants including activated human EGFR viii and PI3KCA H1047R under the control of the radial glial-specific promoter her4.1 in syngeneic tp53 loss-of-function mutant zebrafish. Robust tumor formation was observed prior to 45 days of life, with a gene expression signature similar to human glioblastoma of the mesenchymal subtype, along with a strong inflammatory component. Within early stage tumor lesions, and in an intact and endogenous tumor microenvironment, we visualized infiltration of phagocytic cells, as well as internalization of tumor cells by mpeg1.1:GFP+ microglia/macrophages, suggesting negative regulatory pressure by pro-inflammatory cell types on tumor growth at early stages of glioblastoma initiation in vivo. Furthermore, CRISPR/Cas9-mediated gene targeting of master inflammatory transcription factors irf7 and irf8 led to increased tumor formation in the primary context, while suppression of microglial/macrophage activity led to enhanced tumor cell engraftment following transplantation into otherwise immune competent zebrafish hosts. Altogether, we developed a genetically-relevant model of aggressive human glioblastoma and harnessed the unique advantages of zebrafish including live imaging, high-throughput genetic and chemical manipulations to highlight important tumor suppressive roles for the innate immune system on glioblastoma initiation, with important future significance for therapeutic discovery and optimizations.

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Section: Methodsmentioning

confidence: 99%

A syngeneic spontaneous zebrafish model oftp53-deficient, EGFR^vIII, and PI3KCA^H1047R-driven glioblastoma reveals inhibitory roles for inflammation during tumor initiation and relapsein vivo

Weiss,

D’Amata,

Pearson

et al. 2023

Preprint

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“…The optical clarity of the zebrafish ( Danio rerio ) model makes it uniquely suited for the study of TNLMD as it enables microinjection of cancer cells into LMD-associated anatomical structures while allowing simultaneous fluorescent monitoring of cell proliferation and migration [ 12 , 13 ]. A major advantage of zebrafish xenograft models over rodent models is the availability of pigment-deficient strains, e.g., casper ( roy -/-; nacre -/-), that allow observation of transplanted cancer cells, and the absence until 4–6 weeks after fertilization of an adaptive immune system that could reject foreign implanted cells [ 11 , 14 , 15 ]. The primary conduit for the spread of LMD is the CSF found in the interconnected subarachnoid space and the ventricular system associated with the brain and spinal cord [ 1 , 2 ].…”

Section: Introductionmentioning

confidence: 99%

“…Here, we adapted a zebrafish embryo intraventricular microinjection protocol [ 12 , 15 ] to establish the first zebrafish xenograft embryo model of TNLMD. This model uses the triple-negative breast cancer cell line MDA-MB-231/Luc-RFP, a transgenic reporter breast cancer cell line that allows the detection of cells via red fluorescence expression, which has been used to study cancer xenograft proliferation and migration in zebrafish models [ 20 ].…”

Section: Introductionmentioning

confidence: 99%

Development of a Triple-Negative Breast Cancer Leptomeningeal Disease Model in Zebrafish

Gopal

Monroe

Marudamuthu

et al. 2023

Cells

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Leptomeningeal disease occurs when cancer cells migrate into the ventricles of the brain and spinal cord and then colonize the meninges of the central nervous system. The triple-negative subtype of breast cancer often progresses toward leptomeningeal disease and has a poor prognosis because of limited treatment options. This is due, in part, to a lack of animal models with which to study leptomeningeal disease. Here, we developed a translucent zebrafish casper (roy-/-; nacre-/-) xenograft model of leptomeningeal disease in which fluorescent labeled MDA-MB-231 human triple-negative breast cancer cells are microinjected into the ventricles of zebrafish embryos and then tracked and measured using fluorescent microscopy and multimodal plate reader technology. We then used these techniques to measure tumor area, cell proliferation, and cell death in samples treated with the breast cancer drug doxorubicin and a vehicle control. We monitored MDA-MB-231 cell localization and tumor area, and showed that samples treated with doxorubicin exhibited decreased tumor area and proliferation and increased apoptosis compared to control samples.

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Transplantation of Zebrafish Pediatric Brain Tumors into Immune-competent Hosts for Long-term Study of Tumor Cell Behavior and Drug Response

Cited by 2 publications

References 23 publications

A syngeneic spontaneous zebrafish model oftp53-deficient, EGFR^vIII, and PI3KCA^H1047R-driven glioblastoma reveals inhibitory roles for inflammation during tumor initiation and relapsein vivo

A syngeneic spontaneous zebrafish model oftp53-deficient, EGFR^vIII, and PI3KCA^H1047R-driven glioblastoma reveals inhibitory roles for inflammation during tumor initiation and relapsein vivo

Development of a Triple-Negative Breast Cancer Leptomeningeal Disease Model in Zebrafish

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Transplantation of Zebrafish Pediatric Brain Tumors into Immune-competent Hosts for Long-term Study of Tumor Cell Behavior and Drug Response

Cited by 2 publications

References 23 publications

A syngeneic spontaneous zebrafish model oftp53-deficient, EGFRvIII, and PI3KCAH1047R-driven glioblastoma reveals inhibitory roles for inflammation during tumor initiation and relapsein vivo

A syngeneic spontaneous zebrafish model oftp53-deficient, EGFRvIII, and PI3KCAH1047R-driven glioblastoma reveals inhibitory roles for inflammation during tumor initiation and relapsein vivo

Development of a Triple-Negative Breast Cancer Leptomeningeal Disease Model in Zebrafish

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A syngeneic spontaneous zebrafish model oftp53-deficient, EGFR^vIII, and PI3KCA^H1047R-driven glioblastoma reveals inhibitory roles for inflammation during tumor initiation and relapsein vivo

A syngeneic spontaneous zebrafish model oftp53-deficient, EGFR^vIII, and PI3KCA^H1047R-driven glioblastoma reveals inhibitory roles for inflammation during tumor initiation and relapsein vivo