ARQiv-HTS, a versatile whole-organism screening platform enabling in vivo drug discovery at high-throughput rates

White, David T.; Eroglu, Arife Unal; Wang, Guohua; Zhang, Liyun; Sengupta, Sumitra; Ding, Ding; Rajpurohit, Surendra Kumar; Walker, Steven L.; Ji, Hongkai; Qian, Jiang; Mumm, Jeff S.

doi:10.1038/nprot.2016.142

Cited by 43 publications

(72 citation statements)

References 49 publications

(66 reference statements)

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“…Previously established power analysis methods using ablated and non-ablated controls (White et al, 2016) determined that a sample size of nine larvae was sufficient to detect a 50% neuroprotective effect. For ease of dispensing, microtiter plate formatting, and to account for larval dispensing errors, we increased the sample size to 16 larvae per condition for the primary screen.…”

Section: Establishing a Large-scale Neuroprotectant Screen Using An Imentioning

confidence: 99%

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Large-scale Phenotypic Drug Screen Identifies Neuroprotectants in Zebrafish and Mouse Models of Retinitis Pigmentosa

Zhang

Chen

et al. 2020

Preprint

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Retinitis pigmentosa (RP) and associated inherited retinal diseases (IRDs) are caused by rod photoreceptor degeneration, necessitating therapeutics promoting rod photoreceptor survival. To address this, we tested compounds for neuroprotective effects in zebrafish and mouse RP models, reasoning drugs effective across species may translate better clinically. We first performed a large-scale phenotypic drug screen using a larval zebrafish model of inducible RP. 2,934 compounds, mostly humanapproved drugs, were tested across six concentrations. Statistically, 113 compounds achieved "hit" status. Secondary tests of 42 high-priority hits confirmed eleven lead compounds. Nine leads were then evaluated in mouse RP models, with six exhibiting neuroprotective effects. An analysis of potential mechanisms of action suggested complementary activities. Paired lead compound assays in zebrafish showed additive neuroprotective effects for the majority. These results highlight the value of crossspecies phenotypic drug discovery and suggest combinatorial drug therapies may provide enhanced therapeutic benefits for patients with RP and IRDs.

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Section: Establishing a Large-scale Neuroprotectant Screen Using An Imentioning

confidence: 99%

“…To expand opportunity on this front, we developed a PDD platform enabling quantitative HTS (qHTS; Inglese et al, 2006) in zebrafish (Walker et al, 2012;G. Wang et al, 2015;White et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

Large-scale Phenotypic Drug Screen Identifies Neuroprotectants in Zebrafish and Mouse Models of Retinitis Pigmentosa

Zhang

Chen

et al. 2020

Preprint

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“…In addition, as with humans and rodents, zebrafish possess a BBB which begins to form from just 3dpf in the larvae (Xie, Farage, Sugimoto, & Anand‐Apte, ), allowing one to study the breakdown of barrier permeability, central to conditions such as MS, or to screen for therapeutic compounds capable of crossing the BBB in vivo . Given the promise of using zebrafish for chemical screening, significant effort is currently being invested in automating various aspects of the screening pipeline from mass embryo production systems, automated handling and positioning of embryos (Chang, Pardo‐Martin, Allalou, Wählby, & Yanik, ; Pardo‐Martin et al, ; Pulak, ), to high‐speed imaging, and automated analyses (Wang et al, ; White et al, ). Advances in these areas will help ensure that zebrafish continue to contribute to identifying and developing novel and disease‐relevant therapeutics.…”

Section: Whole‐organism Chemical Screeningmentioning

confidence: 99%

Drug discovery for remyelination and treatment of MS

Cole

Early

Lyons

2017

Glia

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Glia constitute the majority of the cells in our nervous system, yet there are currently no drugs that target glia for the treatment of disease. Given ongoing discoveries of the many roles of glia in numerous diseases of the nervous system, this is likely to change in years to come. Here we focus on the possibility that targeting the oligodendrocyte lineage to promote regeneration of myelin (remyelination) represents a therapeutic strategy for the treatment of the demyelinating disease multiple sclerosis, MS. We discuss how hypothesis driven studies have identified multiple targets and pathways that can be manipulated to promote remyelination in vivo, and how this work has led to the first ever remyelination clinical trials. We also highlight how recent chemical discovery screens have identified a host of small molecule compounds that promote oligodendrocyte differentiation in vitro. Some of these compounds have also been shown to promote myelin regeneration in vivo, with one already being trialled in humans. Promoting oligodendrocyte differentiation and remyelination represents just one potential strategy for the treatment of MS. The pathology of MS is complex, and its complete amelioration may require targeting multiple biological processes in parallel. Therefore, we present an overview of new technologies and models for phenotypic analyses and screening that can be exploited to study complex cell-cell interactions in in vitro and in vivo systems. Such technological platforms will provide insight into fundamental mechanisms and increase capacities for drug-discovery of relevance to glia and currently intractable disorders of the CNS.

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“…Transgenic rho:YFP-NTR larvae were treated with Mtz ± Dex, and regeneration kinetics were analyzed using the ARQiv system, as previously described (20,52). In the preablation Dex regimen, 5-dpf larvae were exposed to Dex (2.5 μM in 0.1% DMSO) for 24 h before Mtz-induced ablation at 6 dpf.…”

Section: Methodsmentioning

confidence: 99%

“…The kinetics of rod cell regeneration was assessed using fluorescent plate reader-based quantification (i.e., automated reporter quantification in vivo, ARQiv) of YFP signal levels in individual larvae (20,51,52). Using this method, we had previously shown full regeneration of rod cells as early as day 5-6 of recovery (20).…”

Section: Microglia/macrophages Are Necessary For Normal Rod Photorecementioning

confidence: 99%

Immunomodulation-accelerated neuronal regeneration following selective rod photoreceptor cell ablation in the zebrafish retina

White¹,

Sengupta²,

Saxena³

et al. 2017

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

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159

218

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Müller glia (MG) function as inducible retinal stem cells in zebrafish, completely repairing the eye after damage. The innate immune system has recently been shown to promote tissue regeneration in which classic wound-healing responses predominate. However, regulatory roles for leukocytes during cellular regeneration-i.e., selective cell-loss paradigms akin to degenerative disease-are less well defined. To investigate possible roles innate immune cells play during retinal cell regeneration, we used intravital microscopy to visualize neutrophil, macrophage, and retinal microglia responses to induced rod photoreceptor apoptosis. Neutrophils displayed no reactivity to rod cell loss. Peripheral macrophage cells responded to rod cell loss, as evidenced by morphological transitions and increased migration, but did not enter the retina. Retinal microglia displayed multiple hallmarks of immune cell activation: increased migration, translocation to the photoreceptor cell layer, proliferation, and phagocytosis of dying cells. To test function during rod cell regeneration, we coablated microglia and rod cells or applied immune suppression and quantified the kinetics of (i) rod cell clearance, (ii) MG/progenitor cell proliferation, and (iii) rod cell replacement. Coablation and immune suppressants applied before cell loss caused delays in MG/progenitor proliferation rates and slowed the rate of rod cell replacement. Conversely, immune suppressants applied after cell loss had been initiated led to accelerated photoreceptor regeneration kinetics, possibly by promoting rapid resolution of an acute immune response. Our findings suggest that microglia control MG responsiveness to photoreceptor loss and support the development of immune-targeted therapeutic strategies for reversing cell loss associated with degenerative retinal conditions. retina | microglia | glucocorticoid | regeneration | macrophage N eurodegenerative diseases are caused by the loss of discrete neuronal cell types. The goal of regenerative medicine is to reverse this process. One strategy for doing so involves harnessing the regenerative potential of endogenous neural stem cells. Unfortunately, although adult neural stem cells exist in the mammalian CNS, innate potentials for neuronal repair remain dormant in the absence of exogenous stimulation. Conversely, many other species are endowed with remarkable capacities for neuronal regeneration. For instance, in the zebrafish eye, retinal Müller glia (MG) cells can dedifferentiate to a stem cell-like state and give rise to progenitors that replace lost neurons and restore visual function (1, 2). Intriguingly, human MG cells are able to give rise to new neurons in culture (3) that can restore visual function when transplanted into mammalian disease models (4), suggesting that human MG retain a stem cell-like capacity for mediating retinal repair. Mechanisms controlling the regenerative potential of MG are therefore of great interest. We and others study zebrafish to learn more about how retinal regeneration is cont...

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ARQiv-HTS, a versatile whole-organism screening platform enabling in vivo drug discovery at high-throughput rates

Cited by 43 publications

References 49 publications

Large-scale Phenotypic Drug Screen Identifies Neuroprotectants in Zebrafish and Mouse Models of Retinitis Pigmentosa

Large-scale Phenotypic Drug Screen Identifies Neuroprotectants in Zebrafish and Mouse Models of Retinitis Pigmentosa

Drug discovery for remyelination and treatment of MS

Immunomodulation-accelerated neuronal regeneration following selective rod photoreceptor cell ablation in the zebrafish retina

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