A versatile, automated and high-throughput drug screening platform for zebrafish embryos

Lubin, Alexandra; Otterstrom, Jason; Hoade, Yvette; Bjedov, Ivana; Stead, Eleanor R.; Whelan, Matt; Gestri, Gaia; Paran, Yael; Payne, Elspeth

doi:10.1242/bio.058513

Cited by 22 publications

(18 citation statements)

References 67 publications

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“…These studies utilized robotic automation to carry out chemical exposure and bright field imaging of unstained zebrafish embryos to identify gross morphological defects. A more recent study used a combination of high-content bright field and fluorescent imaging followed by artificial intelligence-automated cell counting analysis to enumerate hematopoietic stem and progenitor cells in the tails of embryos, which comported with manual cell counts (Lubin et al, 2021). However, the analysis software is proprietary, reducing versatility, and accessibility of the assay.…”

Section: Discussionmentioning

confidence: 99%

High‐throughput detection of craniofacial defects in fluorescent zebrafish

Everson

Tseng

Eberhart

2022

Birth Defects Research

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Losses and malformations of cranial neural crest cell (cNCC) derivatives are a hallmark of several common brain and face malformations. Nevertheless, the etiology of these cNCC defects remains unknown for many cases, suggesting a complex basis involving interactions between genetic and/or environmental factors. However, the sheer number of possible factors (thousands of genes and hundreds of thousands of toxicants) has hindered identification of specific interactions. Here, we develop a high‐throughput analysis that will enable faster identification of multifactorial interactions in the genesis of craniofacial defects. Zebrafish embryos expressing a fluorescent marker of cNCCs (fli1:EGFP) were exposed to a pathway inhibitor standard or environmental toxicant, and resulting changes in fluorescence were measured in high‐throughput using a fluorescent microplate reader to approximate cNCC losses. Embryos exposed to the environmental Hedgehog pathway inhibitor piperonyl butoxide (PBO), a Hedgehog pathway inhibitor standard, or alcohol (ethanol) exhibited reduced fli1:EGFP fluorescence at one day post fertilization, which corresponded with craniofacial defects at five days post fertilization. Combining PBO and alcohol in a co‐exposure paradigm synergistically reduced fluorescence, demonstrating a multifactorial interaction. Using pathway reporter transgenics, we show that the plate reader assay is sensitive at detecting alterations in Hedgehog signaling, a critical regulator of craniofacial development. We go on to demonstrate that this technique readily detects defects in other important cell types, namely neurons. Together, these findings demonstrate this novel in vivo platform can predict developmental abnormalities and multifactorial interactions in high‐throughput.

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Section: Discussionmentioning

confidence: 99%

High‐throughput detection of craniofacial defects in fluorescent zebrafish

Everson

Tseng

Eberhart

2022

Birth Defects Research

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“…123 Fortunately, there has been a significant improvement in the development of automated screening technologies in the last few years, which contribute to address these limitations. [124][125][126] Dyballa et al 127 have recently described an exceptionally promising platform for zebrafish cardiovascular screens. This system relies on the Vertebrate Automated Screening Technology, which aspirates embryos from a multi-well plate, loads them head-first in a capillary tube, and rotates the embryo to the correct orientation for imaging.…”

Section: González-rosamentioning

confidence: 99%

Zebrafish Models of Cardiac Disease: From Fortuitous Mutants to Precision Medicine

González‐Rosa

2022

Circulation Research

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Heart disease is the leading cause of death worldwide. Despite decades of research, most heart pathologies have limited treatments, and often the only curative approach is heart transplantation. Thus, there is an urgent need to develop new therapeutic approaches for treating cardiac diseases. Animal models that reproduce the human pathophysiology are essential to uncovering the biology of diseases and discovering therapies. Traditionally, mammals have been used as models of cardiac disease, but the cost of generating and maintaining new models is exorbitant, and the studies have very low throughput. In the last decade, the zebrafish has emerged as a tractable model for cardiac diseases, owing to several characteristics that made this animal popular among developmental biologists. Zebrafish fertilization and development are external; embryos can be obtained in high numbers, are cheap and easy to maintain, and can be manipulated to create new genetic models. Moreover, zebrafish exhibit an exceptional ability to regenerate their heart after injury. This review summarizes 25 years of research using the zebrafish to study the heart, from the classical forward screenings to the contemporary methods to model mutations found in patients with cardiac disease. We discuss the advantages and limitations of this model organism and introduce the experimental approaches exploited in zebrafish, including forward and reverse genetics and chemical screenings. Last, we review the models used to induce cardiac injury and essential ideas derived from studying natural regeneration. Studies using zebrafish have the potential to accelerate the discovery of new strategies to treat cardiac diseases.

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“…Through the new platform, the group investigated the chemosensitivity of 5-fluorouracil, cisplatin, docetaxel, and doxorubicin in gastric cancer patients ( 53 ). Zebrafish cancer models, when combined with large-scale genetic screens and drug discovery platforms, have provided valuable insights into the clonal evolution and heterogeneity of tumors, drug resistance to therapy, invasion, metastasis, hematopoiesis, and transplantation of stem cells ( 54 – 61 ).…”

Section: Alternative Hosts In Gc Patient-derived Modelsmentioning

confidence: 99%

Patient-Derived Xenograft: A More Standard “Avatar” Model in Preclinical Studies of Gastric Cancer

Zeng

et al. 2022

Front. Oncol.

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Despite advances in diagnosis and treatment, gastric cancer remains the third most common cause of cancer-related death in humans. The establishment of relevant animal models of gastric cancer is critical for further research. Due to the complexity of the tumor microenvironment and the genetic heterogeneity of gastric cancer, the commonly used preclinical animal models fail to adequately represent clinically relevant models of gastric cancer. However, patient-derived models are able to replicate as much of the original inter-tumoral and intra-tumoral heterogeneity of gastric cancer as possible, reflecting the cellular interactions of the tumor microenvironment. In addition to implanting patient tissues or primary cells into immunodeficient mouse hosts for culture, the advent of alternative hosts such as humanized mouse hosts, zebrafish hosts, and in vitro culture modalities has also facilitated the advancement of gastric cancer research. This review highlights the current status, characteristics, interfering factors, and applications of patient-derived models that have emerged as more valuable preclinical tools for studying the progression and metastasis of gastric cancer.

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A versatile, automated and high-throughput drug screening platform for zebrafish embryos

Cited by 22 publications

References 67 publications

High‐throughput detection of craniofacial defects in fluorescent zebrafish

High‐throughput detection of craniofacial defects in fluorescent zebrafish

Zebrafish Models of Cardiac Disease: From Fortuitous Mutants to Precision Medicine

Patient-Derived Xenograft: A More Standard “Avatar” Model in Preclinical Studies of Gastric Cancer

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