High-throughput screening of human induced pluripotent stem cell-derived brain organoids

Durens, Madel; Nestor, Jonathan E.; Williams, Madeline; Herold, Kevin; Niescier, Robert F.; Lunden, Jason W; Phillips, André W.; Lin, Yu Chih; Dykxhoorn, Derek M.; Nestor, Michael W.

doi:10.1016/j.jneumeth.2020.108627

Cited by 67 publications

(61 citation statements)

References 23 publications

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“…Now sensory neurons can be derived from hiPSCs, and they can be recruited to develop functional assay of neuronal excitability to be then employed for the evaluation of the phenotypic effects of a smalltargeted validation compound subset [33]. Lately, hiPSCs have set the foundation for the development of a 3D human stem cell platform to screen cortical organoids with high-throughput screening (HTS) [34]. By assembling the reprogramming technology with the use of high content imaging, it was possible to obtain Serum-Free Embryoid Bodies (SFEBs) and then to determine neurite outgrowth and cellular composition.…”

Section: Hipscs As a Drug Discovery Devicementioning

confidence: 99%

“…By assembling the reprogramming technology with the use of high content imaging, it was possible to obtain Serum-Free Embryoid Bodies (SFEBs) and then to determine neurite outgrowth and cellular composition. In this way, the detection of neurite morphology and multi-electrode array analysis were carried out [34]. This approach turned out to be a valid idea for countering the experimental variability common in 3D cultures; this study makes the conjunction between SFEBs and HTS the baseline for phenotypic drug screening [34].…”

Section: Hipscs As a Drug Discovery Devicementioning

confidence: 99%

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Application of CRISPR/Cas9 to human-induced pluripotent stem cells: from gene editing to drug discovery

et al. 2020

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Human-induced pluripotent stem cells (hiPSCs) and CRISPR/Cas9 gene editing system represent two instruments of basic and translational research, which both allow to acquire deep insight about the molecular bases of many diseases but also to develop pharmacological research. This review is focused to draw up the latest technique of gene editing applied on hiPSCs, exploiting some of the genetic manipulation directed to the discovery of innovative therapeutic strategies. There are many expediencies provided by the use of hiPSCs, which can represent a disease model clinically relevant and predictive, with a great potential if associated to CRISPR/Cas9 technology, a gene editing tool powered by ease and precision never seen before. Here, we describe the possible applications of CRISPR/Cas9 to hiPSCs: from drug development to drug screening and from gene therapy to the induction of the immunological response to specific virus infection, such as HIV and SARS-Cov-2.

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Section: Hipscs As a Drug Discovery Devicementioning

confidence: 99%

Section: Hipscs As a Drug Discovery Devicementioning

confidence: 99%

Application of CRISPR/Cas9 to human-induced pluripotent stem cells: from gene editing to drug discovery

et al. 2020

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“…Regarding the adaptation of the methodology for use in high-throughput screening, Durens et al have described a method for multiplexing MEA recordings and Ca2+ imaging to examine local microcircuits in 3D brain organoids and assess inter-experimental consistency, necessary for drug screening [ 160 ]. Examination of such circuit-based mechanisms in hiPSC-derived systems would be of great interest and importance for the development of new and more efficient drugs.…”

Section: Functional Assays For Pd Studies and Drug Screening In Himentioning

confidence: 99%

Patient-Derived Induced Pluripotent Stem Cell-Based Models in Parkinson’s Disease for Drug Identification

Kouroupi

Antoniou

Prodromidou

et al. 2020

IJMS

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Parkinson’s disease (PD) is a common progressive neurodegenerative disorder characterized by loss of striatal-projecting dopaminergic neurons of the ventral forebrain, resulting in motor and cognitive deficits. Despite extensive efforts in understanding PD pathogenesis, no disease-modifying drugs exist. Recent advances in cell reprogramming technologies have facilitated the generation of patient-derived models for sporadic or familial PD and the identification of early, potentially triggering, pathological phenotypes while they provide amenable systems for drug discovery. Emerging developments highlight the enhanced potential of using more sophisticated cellular systems, including neuronal and glial co-cultures as well as three-dimensional systems that better simulate the human pathophysiology. In combination with high-throughput high-content screening technologies, these approaches open new perspectives for the identification of disease-modifying compounds. In this review, we discuss current advances and the challenges ahead in the use of patient-derived induced pluripotent stem cells for drug discovery in PD. We address new concepts implicating non-neuronal cells in disease pathogenesis and highlight the necessity for functional assays, such as calcium imaging and multi-electrode array recordings, to predict drug efficacy. Finally, we argue that artificial intelligence technologies will be pivotal for analysis of the large and complex data sets obtained, becoming game-changers in the process of drug discovery.

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“…As a consequence, hiPSCs can be obtained from every individual and similarly, it is possible to differentiate personalized cerebral (and other) organoids. COs could be on their way to become an indispensable technology for drug screening experiments [4], biomarker identi cation, and drug treatment response analyses [5] for different elds of research, including neurodevelopment, neurodegeneration, and psychiatric disorders and the list is continuously growing. Despite these amazing features, the cerebral organoid technology is currently facing challenges, e.g.…”

Section: Introductionmentioning

confidence: 99%

Gene Edited Fluorescent Cerebral Organoids to Study Human Brain Function and Disease

Müeller

Bachmann

Villarejo

et al. 2021

Preprint

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Cerebral organoids are a promising model to study human brain function and disease, though the high inter-organoid variability of the mini-brains is still challenging. To overcome this limitation, we introduce the method of labeled mixed organoids generated from two different human induced pluripotent stem cell (hiPSC) lines, which enables the identification of cells from different origin within a single organoid. The method combines a gene editing workflow and subsequent organoid differentiation and offers a unique tool to study gene function in a complex human 3D tissue-like model. Using a CRISPR/Cas9 gene editing approach, different fluorescent proteins were fused to β-actin or lamin B1 in hiPSCs and subsequently used as a marker to identify each cell line. Mixtures of differently edited cells were seeded to induce embryoid body formation and cerebral organoid differentiation. As a consequence, the development of the 3D tissue was detectable by live confocal fluorescence microscopy and immunofluorescence staining in fixed samples. Analysis of mixed organoids allowed the identification and examination of specifically labeled cells in the organoid that belong to each of the two hiPSC donor lines. We demonstrate that a direct comparison of the individual cells is possible by having the edited and the control (or the two differentially labeled) cells within the same organoid, and thus the mixed organoids overcome the inter-organoid inhomogeneity limitations. The approach aims to pave the way for the reliable analysis of human genetic disorders by the use of organoids and to fundamentally understand the molecular mechanisms underlying pathological conditions.

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High-throughput screening of human induced pluripotent stem cell-derived brain organoids

Cited by 67 publications

References 23 publications

Application of CRISPR/Cas9 to human-induced pluripotent stem cells: from gene editing to drug discovery

Application of CRISPR/Cas9 to human-induced pluripotent stem cells: from gene editing to drug discovery

Patient-Derived Induced Pluripotent Stem Cell-Based Models in Parkinson’s Disease for Drug Identification

Gene Edited Fluorescent Cerebral Organoids to Study Human Brain Function and Disease

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