Large-Scale Phenotype-Based Antiepileptic Drug Screening in a Zebrafish Model of Dravet Syndrome

In the last decade, different research groups in the academic setting have developed induced pluripotent stem cell-based protocols to generate three-dimensional, multicellular, neural organoids. Their use to model brain biology, early neural development, and human diseases has provided new insights into the pathophysiology of neuropsychiatric and neurological disorders, including microcephaly, autism, Parkinson’s disease, and Alzheimer’s disease. However, the adoption of organoid technology for large-scale drug screening in the industry has been hampered by challenges with reproducibility, scalability, and translatability to human disease. Potential technical solutions to expand their use in drug discovery pipelines include Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) to create isogenic models, single-cell RNA sequencing to characterize the model at a cellular level, and machine learning to analyze complex data sets. In addition, high-content imaging, automated liquid handling, and standardized assays represent other valuable tools toward this goal. Though several open issues still hamper the full implementation of the organoid technology outside academia, rapid progress in this field will help to prompt its translation toward large-scale drug screening for neurological disorders.

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Preliminary Results Regarding Sleep in a Zebrafish Model of Autism Spectrum Disorder

Brain Sciences ◽

10.3390/brainsci11050556 ◽

2021 ◽

Vol 11 (5) ◽

pp. 556

Author(s):

Madalina Andreea Robea ◽

Alin Ciobica ◽

Alexandrina-Stefania Curpan ◽

Gabriel Plavan ◽

Stefan Strungaru ◽

...

Keyword(s):

Autism Spectrum Disorder ◽

Valproic Acid ◽

Social Behavior ◽

Antiepileptic Drug ◽

Alternative Model ◽

Neurological Diseases ◽

Autism Spectrum ◽

Spectrum Disorder ◽

Zebrafish Model ◽

Developmental Defects

Autism spectrum disorder (ASD) is one of the most salient developmental neurological diseases and remarkable similarities have been found between humans and model animals of ASD. A common method of inducing ASD in zebrafish is by administrating valproic acid (VPA), which is an antiepileptic drug that is strongly linked with developmental defects in children. In the present study we replicated and extended the findings of VPA on social behavior in zebrafish by adding several sleep observations. Juvenile zebrafish manifested hyperactivity and an increase in ASD-like social behaviors but, interestingly, only exhibited minimal alterations in sleep. Our study confirmed that VPA can generate specific ASD symptoms, indicating that the zebrafish is an alternative model in this field of research.

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TMOD-29. STANDARDIZED GENERATION OF TUMOR-ORGANOIDS AS NOVEL DRUG SCREENING MODEL IN MENINGIOMA

Neuro-Oncology ◽

10.1093/neuonc/noab196.890 ◽

2021 ◽

Vol 23 (Supplement_6) ◽

pp. vi221-vi222

Author(s):

Gerhard Jungwirth ◽

Tao Yu ◽

Cao Junguo ◽

Catharina Lotsch ◽

Andreas Unterberg ◽

...

Keyword(s):

Drug Screening ◽

Cancer Biology ◽

Drug Testing ◽

Large Scale ◽

Ex Vivo ◽

Cell Types ◽

Cellular Heterogeneity ◽

Drug Responses ◽

Novel Drug ◽

Tumor Organoids

Abstract Tumor-organoids (TOs) are novel, complex three-dimensional ex vivo tissue cultures that under optimal conditions accurately reflect genotype and phenotype of the original tissue with preserved cellular heterogeneity and morphology. They may serve as a new and exciting model for studying cancer biology and directing personalized therapies. The aim of our study was to establish TOs from meningioma (MGM) and to test their usability for large-scale drug screenings. We were capable of forming several hundred TO equal in size by controlled reaggregation of freshly prepared single cell suspension of MGM tissue samples. In total, standardized TOs from 60 patients were formed, including eight grade II and three grade III MGMs. TOs reaggregated within 3 days resulting in a reducted diameter by 50%. Thereafter, TO size remained stable throughout a 14 days observation period. TOs consisted of largely viable cells, whereas dead cells were predominantly found outside of the organoid. H&E stainings confirmed the successful establishment of dense tissue-like structures. Next, we assessed the suitability and reliability of TOs for a robust large-scale drug testing by employing nine highly potent compounds, derived from a drug screening performed on several MGM cell lines. First, we tested if drug responses depend on TO size. Interestingly, drug responses to these drugs remained identical independent of their sizes. Based on a sufficient representation of low abundance cell types such as T-cells and macrophages an overall number of 25.000 cells/TO was selected for further experiments revealing FDA-approved HDAC inhibitors as highly effective drugs in most of the TOs with a mean z-AUC score of -1.33. Taken together, we developed a protocol to generate standardized TO from MGM containing low abundant cell types of the tumor microenvironment in a representative manner. Robust and reliable drug responses suggest patient-derived TOs as a novel drug testing model in meningioma research.

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Testing Tuberculosis Drug Efficacy in a Zebrafish High-Throughput Translational Medicine Screen

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.03588-14 ◽

2014 ◽

Vol 59 (2) ◽

pp. 753-762 ◽

Cited By ~ 36

Author(s):

Anita Ordas ◽

Robert-Jan Raterink ◽

Fraser Cunningham ◽

Hans J. Jansen ◽

Malgorzata I. Wiweger ◽

...

Keyword(s):

High Throughput ◽

Drug Screening ◽

Drug Efficacy ◽

Drug Uptake ◽

Spectrometric Analysis ◽

Major Advance ◽

Zebrafish Model ◽

Content Type ◽

Screening Platform

ABSTRACTThe translational value of zebrafish high-throughput screens can be improved when more knowledge is available on uptake characteristics of potential drugs. We investigated reference antibiotics and 15 preclinical compounds in a translational zebrafish-rodent screening system for tuberculosis. As a major advance, we have developed a new tool for testing drug uptake in the zebrafish model. This is important, because despite the many applications of assessing drug efficacy in zebrafish research, the current methods for measuring uptake using mass spectrometry do not take into account the possible adherence of drugs to the larval surface. Our approach combines nanoliter sampling from the yolk using a microneedle, followed by mass spectrometric analysis. To date, no single physicochemical property has been identified to accurately predict compound uptake; our method offers a great possibility to monitor how any novel compound behaves within the system. We have correlated the uptake data with high-throughput drug-screening data fromMycobacterium marinum-infected zebrafish larvae. As a result, we present an improved zebrafish larva drug-screening platform which offers new insights into drug efficacy and identifies potential false negatives and drugs that are effective in zebrafish and rodents. We demonstrate that this improved zebrafish drug-screening platform can complement conventional models ofin vivoMycobacterium tuberculosis-infected rodent assays. The detailed comparison of two vertebrate systems, fish and rodent, may give more predictive value for efficacy of drugs in humans.

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Large-Scale Production of Human iPSC-Derived Macrophages for Drug Screening

International Journal of Molecular Sciences ◽

10.3390/ijms21134808 ◽

2020 ◽

Vol 21 (13) ◽

pp. 4808 ◽

Cited By ~ 3

Author(s):

Simon Gutbier ◽

Florian Wanke ◽

Nadine Dahm ◽

Anna Rümmelin ◽

Silke Zimmermann ◽

...

Keyword(s):

Drug Screening ◽

Large Scale ◽

Neoplastic Cell ◽

Leukemic Cells ◽

High Yield ◽

Scale Production ◽

Large Scale Production ◽

Compound Screening ◽

Health And Disease ◽

Induced Pluripotent

Tissue-resident macrophages are key players in inflammatory processes, and their activation and functionality are crucial in health and disease. Numerous diseases are associated with alterations in homeostasis or dysregulation of the innate immune system, including allergic reactions, autoimmune diseases, and cancer. Macrophages are a prime target for drug discovery due to their major regulatory role in health and disease. Currently, the main sources of macrophages used for therapeutic compound screening are primary cells isolated from blood or tissue or immortalized or neoplastic cell lines (e.g., THP-1). Here, we describe an improved method to employ induced pluripotent stem cells (iPSCs) for the high-yield, large-scale production of cells resembling tissue-resident macrophages. For this, iPSC-derived macrophage-like cells are thoroughly characterized to confirm their cell identity and thus their suitability for drug screening purposes. These iPSC-derived macrophages show strong cellular identity with primary macrophages and recapitulate key functional characteristics, including cytokine release, phagocytosis, and chemotaxis. Furthermore, we demonstrate that genetic modifications can be readily introduced at the macrophage-like progenitor stage in order to interrogate drug target-relevant pathways. In summary, this novel method overcomes previous shortcomings with primary and leukemic cells and facilitates large-scale production of genetically modified iPSC-derived macrophages for drug screening applications.

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A vascularized and perfused organ-on-a-chip platform for large-scale drug screening applications

Lab on a Chip ◽

10.1039/c6lc01422d ◽

2017 ◽

Vol 17 (3) ◽

pp. 511-520 ◽

Cited By ~ 109

Author(s):

Duc T. T. Phan ◽

Xiaolin Wang ◽

Brianna M. Craver ◽

Agua Sobrino ◽

Da Zhao ◽

...

Keyword(s):

Drug Screening ◽

Large Scale ◽

Drug Efficacy ◽

Toxicity Screening ◽

Organ On A Chip

A vascularized, perfused organ-on-a-chip platform suitable for large-scale drug efficacy/toxicity screening.

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From Omics to Drug Metabolism and High Content Screen of Natural Product in Zebrafish: A New Model for Discovery of Neuroactive Compound

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2012/605303 ◽

2012 ◽

Vol 2012 ◽

pp. 1-20 ◽

Cited By ~ 21

Author(s):

Ming Wai Hung ◽

Zai Jun Zhang ◽

Shang Li ◽

Benson Lei ◽

Shuai Yuan ◽

...

Keyword(s):

Drug Metabolism ◽

Drug Screening ◽

Environmental Science ◽

Pharmacological Action ◽

Zebrafish Model ◽

Lead Compounds ◽

Efficacy Parameters

The zebrafish (Danio rerio) has recently become a common model in the fields of genetics, environmental science, toxicology, and especially drug screening. Zebrafish has emerged as a biomedically relevant model forin vivohigh content drug screening and the simultaneous determination of multiple efficacy parameters, including behaviour, selectivity, and toxicity in the content of the whole organism. A zebrafish behavioural assay has been demonstrated as a novel, rapid, and high-throughput approach to the discovery of neuroactive, psychoactive, and memory-modulating compounds. Recent studies found a functional similarity of drug metabolism systems in zebrafish and mammals, providing a clue with why some compounds are active in zebrafishin vivobut notin vitro, as well as providing grounds for the rationales supporting the use of a zebrafish screen to identify prodrugs. Here, we discuss the advantages of the zebrafish model for evaluating drug metabolism and the mode of pharmacological action with the emerging omics approaches. Why this model is suitable for identifying lead compounds from natural products for therapy of disorders with multifactorial etiopathogenesis and imbalance of angiogenesis, such as Parkinson's disease, epilepsy, cardiotoxicity, cerebral hemorrhage, dyslipidemia, and hyperlipidemia, is addressed.

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