Single-cell imaging of normal and malignant cell engraftment into optically clear <i>prkdc</i>-null SCID zebrafish

Moore, John C.; Tang, Qin; Yordán, Nora Torres; Moore, Finola; Garcia, Elaine G.; Lobbardi, Riadh; Ramakrishnan, Ashwin; Marvin, Dieuwke L.; Anselmo, Anthony; Sadreyev, Ruslan I.; Langenau, David M.

doi:10.1084/jem.20160378

Cited by 67 publications

(83 citation statements)

References 60 publications

(104 reference statements)

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“…As observed with cell lines (SW480), patient samples (e.g., zPDX#3) also vary in their engraftment efficiency (being as low as <50%). To overcome possible low implantation rates, we may increase the number of injected fish and use immune-compromised strains (43,44) to dampen the possibility of rejection. As a proof of concept experiments, we also treated zPDX with FOLFOX and were able to anticipate relapse/no relapse within 3 m to 6 m after surgery in four out of five patients.…”

Section: Discussionmentioning

confidence: 99%

Single-cell functional and chemosensitive profiling of combinatorial colorectal therapy in zebrafish xenografts

Fior

Póvoa

Mendes

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

263

368

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Cancer is as unique as the person fighting it. With the exception of a few biomarker-driven therapies, patients go through rounds of trial-and-error approaches to find the best treatment. Using patient-derived cell lines, we show that zebrafish larvae xenotransplants constitute a fast and highly sensitive in vivo model for differential therapy response, with resolution to reveal intratumor functional cancer heterogeneity. We screened international colorectal cancer therapeutic guidelines and determined distinct functional tumor behaviors (proliferation, metastasis, and angiogenesis) and differential sensitivities to standard therapy. We observed a general higher sensitivity to FOLFIRI [5-fluorouracil(FU)+irinotecan+folinic acid] than to FOLFOX (5-FU+oxaliplatin+folinic acid), not only between isogenic tumors but also within the same tumor. We directly compared zebrafish xenografts with mouse xenografts and show that relative sensitivities obtained in zebrafish are maintained in the rodent model. Our data also illustrate how mutations can provide proliferation advantages in relation to KRASWT and how chemotherapy can unbalance this advantage, selecting for a minor clone resistant to chemotherapy. Zebrafish xenografts provide remarkable resolution to measure Cetuximab sensitivity. Finally, we demonstrate the feasibility of using primary patient samples to generate zebrafish patient-derived xenografts (zPDX) and provide proof-of-concept experiments that compare response to chemotherapy and biological therapies between patients and zPDX. Altogether, our results suggest that zebrafish larvae xenografts constitute a promising fast assay for precision medicine, bridging the gap between genotype and phenotype in an in vivo setting.

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Section: Discussionmentioning

confidence: 99%

Single-cell functional and chemosensitive profiling of combinatorial colorectal therapy in zebrafish xenografts

Fior

Póvoa

Mendes

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

263

368

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“…The rag2 450fs/+ ;casper and prkdc D3612fs/+ ;casper lines were maintained as heterozygotes. Heterozygotes were crossed, and homozygotes, which were used in experiments, were identified using previously described genotyping protocols [ 28 , 30 ]. Young adult zebrafish used for experiments were between 6 and 10 weeks old and were housed at a density of 15 fish per 3 L tank.…”

Section: Methodsmentioning

confidence: 99%

“…Without these two types of pigment cells, zebrafish are transparent even as adults, eliminating the need for any further manipulation of the animal prior to experimentation. Casper fish have been used to image the clonal heterogeneity [ 27 , 28 ] and neovascularization [ 27 ] of transplanted primary tumors. Casper fish have also been used as a quantitative system to study metastasis using fluorescence as a readout [ 29 ].…”

Section: Introductionmentioning

confidence: 99%

Intravital imaging of metastasis in adult Zebrafish

Benjamin¹,

Hynes²

2017

BMC Cancer

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BackgroundMetastasis is a major clinical problem whose biology is not yet fully understood. This lack of understanding is especially true for the events at the metastatic site, which include arrest, extravasation, and growth into macrometastases. Intravital imaging is a powerful technique that has shown great promise in increasing our understanding of these events. To date, most intravital imaging studies have been performed in mice, which has limited its adoption. Zebrafish are also a common system for the intravital imaging of metastasis. However, as imaging in embryos is technically simpler, relatively few studies have used adult zebrafish to study metastasis and none have followed individual cells at the metastatic site over time. The aim of this study was to demonstrate that adult casper zebrafish offer a convenient model system for performing intravital imaging of the metastatic site over time with single-cell resolution.MethodsZMEL1 zebrafish melanoma cells were injected into 6 to 10-week-old casper fish using an intravenous injection protocol. Because casper fish are transparent even as adults, they could be imaged without surgical intervention. Individual cells were followed over the course of 2 weeks as they arrested, extravasated, and formed macroscopic metastases.ResultsOur injection method reliably delivered cells into circulation and led to the formation of tumors in multiple organs. Cells in the skin and sub-dermal muscle could be imaged at high resolution over 2 weeks using confocal microscopy. Arrest was visualized and determined to be primarily due to size restriction. Following arrest, extravasation was seen to occur between 1 and 6 days post-injection. Once outside of the vasculature, cells were observed migrating as well as forming protrusions.Conclusions Casper fish are a useful model for studying the events at the metastatic site using intravital imaging. The protocols described in this study are relatively simple. Combined with the reasonably low cost of zebrafish, they offer to increase access to intravital imaging.Electronic supplementary materialThe online version of this article (10.1186/s12885-017-3647-0) contains supplementary material, which is available to authorized users.

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“…For some slowly growing primary tumor cells, the short experimental setup proposed for PDXzs might actually be disadvantageous. Here, several immunocompromised zebrafish strains like rag2 E450fs , jak3 P369fs , prkdc D3612fs , and zap70 y442 , which can be combined with optically clear mutant strains like casper, will help to overcome adaptive immune response and imaging problems associated with performing xenograft studies in juvenile zebrafish ( 21 , 113 – 115 ).…”

Section: Toward Pdx In Zebrafishmentioning

confidence: 99%

Quo natas, Danio?—Recent Progress in Modeling Cancer in Zebrafish

et al. 2017

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Over the last decade, zebrafish has proven to be a powerful model in cancer research. Zebrafish form tumors that histologically and genetically resemble human cancers. The live imaging and cost-effective compound screening possible with zebrafish especially complement classic mouse cancer models. Here, we report recent progress in the field, including genetically engineered zebrafish cancer models, xenotransplantation of human cancer cells into zebrafish, promising approaches toward live investigation of the tumor microenvironment, and identification of therapeutic strategies by performing compound screens on zebrafish cancer models. Given the recent advances in genome editing, personalized zebrafish cancer models are now a realistic possibility. In addition, ongoing automation will soon allow high-throughput compound screening using zebrafish cancer models to be part of preclinical precision medicine approaches.

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Single-cell imaging of normal and malignant cell engraftment into optically clear prkdc-null SCID zebrafish

Abstract: Moore and colleagues present new strains of optically clear immune-deficient zebrafish that allow for dynamic imaging of regeneration and tumor progression at single-cell resolution in live animals.

Cited by 67 publications

References 60 publications

Single-cell functional and chemosensitive profiling of combinatorial colorectal therapy in zebrafish xenografts

Single-cell functional and chemosensitive profiling of combinatorial colorectal therapy in zebrafish xenografts

Intravital imaging of metastasis in adult Zebrafish

Quo natas, Danio?—Recent Progress in Modeling Cancer in Zebrafish

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