scholarly journals Autobioluminescent Cellular Models for Enhanced Drug Discovery

10.5772/65062 ◽  
2016 ◽  
Author(s):  
Tingting Xu ◽  
Michael Conway ◽  
Ashley Frank ◽  
Amelia Brumbaugh ◽  
Steven Ripp ◽  
...  
Keyword(s):  
Drug Discovery ◽  
Cellular Models

Recommended Guidelines for Developing, Qualifying, and Implementing Complex In Vitro Models (CIVMs) for Drug Discovery

2020 ◽  
Vol 25 (10) ◽  
pp. 1174-1190
Author(s):  
Jason E. Ekert ◽  
Julianna Deakyne ◽  
Philippa Pribul-Allen ◽  
Rebecca Terry ◽  
Christopher Schofield ◽  
...  
Keyword(s):  
Drug Discovery ◽  
Early Stage ◽  
Cell Types ◽  
In Vitro Models ◽  
Success Rates ◽  
Lead Discovery ◽  
Cellular Models ◽  
Discovery Phase ◽  
Early Drug

The pharmaceutical industry is continuing to face high research and development (R&D) costs and low overall success rates of clinical compounds during drug development. There is an increasing demand for development and validation of healthy or disease-relevant and physiological human cellular models that can be implemented in early-stage discovery, thereby shifting attrition of future therapeutics to a point in discovery at which the costs are significantly lower. There needs to be a paradigm shift in the early drug discovery phase (which is lengthy and costly), away from simplistic cellular models that show an inability to effectively and efficiently reproduce healthy or human disease-relevant states to steer target and compound selection for safety, pharmacology, and efficacy questions. This perspective article covers the various stages of early drug discovery from target identification (ID) and validation to the hit/lead discovery phase, lead optimization, and preclinical safety. We outline key aspects that should be considered when developing, qualifying, and implementing complex in vitro models (CIVMs) during these phases, because criteria such as cell types (e.g., cell lines, primary cells, stem cells, and tissue), platform (e.g., spheroids, scaffolds or hydrogels, organoids, microphysiological systems, and bioprinting), throughput, automation, and single and multiplexing endpoints will vary. The article emphasizes the need to adequately qualify these CIVMs such that they are suitable for various applications (e.g., context of use) of drug discovery and translational research. The article ends looking to the future, in which there is an increase in combining computational modeling, artificial intelligence and machine learning (AI/ML), and CIVMs.

The Use of Real-Time Cell Analyzer Technology in Drug Discovery: Defining Optimal Cell Culture Conditions and Assay Reproducibility with Different Adherent Cellular Models

2011 ◽  
Vol 16 (6) ◽  
pp. 575-587 ◽  
Author(s):  
Franck A. Atienzar ◽  
Karen Tilmant ◽  
Helga H. Gerets ◽  
Gaelle Toussaint ◽  
Sebastien Speeckaert ◽  
...  
Keyword(s):  
Quality Control ◽  
Drug Discovery ◽  
Real Time ◽  
Culture Conditions ◽  
Label Free ◽  
Kinetic Measurements ◽  
Cell Index ◽  
Cellular Models ◽  
Index Data ◽  
Cell Analyzer

The use of impedance-based label-free technology applied to drug discovery is nowadays receiving more and more attention. Indeed, such a simple and noninvasive assay that interferes minimally with cell morphology and function allows one to perform kinetic measurements and to obtain information on proliferation, migration, cytotoxicity, and receptor-mediated signaling. The objective of the study was to further assess the usefulness of a real-time cell analyzer (RTCA) platform based on impedance in the context of quality control and data reproducibility. The data indicate that this technology is useful to determine the best coating and cellular density conditions for different adherent cellular models including hepatocytes, cardiomyocytes, fibroblasts, and hybrid neuroblastoma/neuronal cells. Based on 31 independent experiments, the reproducibility of cell index data generated from HepG2 cells exposed to DMSO and to Triton X-100 was satisfactory, with a coefficient of variation close to 10%. Cell index data were also well reproduced when cardiomyocytes and fibroblasts were exposed to 21 compounds three times (correlation >0.91, p < 0.0001). The data also show that a cell index decrease is not always associated with cytotoxicity effects and that there are some confounding factors that can affect the analysis. Finally, another drawback is that the correlation analysis between cellular impedance measurements and classical toxicity endpoints has been performed on a limited number of compounds. Overall, despite some limitations, the RTCA technology appears to be a powerful and reliable tool in drug discovery because of the reasonable throughput, rapid and efficient performance, technical optimization, and cell quality control.

scholarly journals Functional patient-derived cellular models for neuropsychiatric drug discovery

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Santiago G. Lago ◽  
Jakub Tomasik ◽  
Sabine Bahn
Keyword(s):  
Drug Discovery ◽  
Mononuclear Cells ◽  
Induced Pluripotent Stem Cell ◽  
Functional Responses ◽  
Peripheral Blood Mononuclear ◽  
Disease Heterogeneity ◽  
Drug Candidates ◽  
Cellular Models ◽  
Novel Strategy ◽  
Novel Drug

AbstractMental health disorders are a leading cause of disability worldwide. Challenges such as disease heterogeneity, incomplete characterization of the targets of existing drugs and a limited understanding of functional interactions of complex genetic risk loci and environmental factors have compromised the identification of novel drug candidates. There is a pressing clinical need for drugs with new mechanisms of action which address the lack of efficacy and debilitating side effects of current medications. Here we discuss a novel strategy for neuropsychiatric drug discovery which aims to address these limitations by identifying disease-related functional responses (‘functional cellular endophenotypes’) in a variety of patient-derived cells, such as induced pluripotent stem cell (iPSC)-derived neurons and organoids or peripheral blood mononuclear cells (PBMCs). Disease-specific alterations in cellular responses can subsequently yield novel drug screening targets and drug candidates. We discuss the potential of this approach in the context of recent advances in patient-derived cellular models, high-content single-cell screening of cellular networks and changes in the diagnostic framework of neuropsychiatric disorders.

Operationalizing the Use of Biofabricated Tissue Models as Preclinical Screening Platforms for Drug Discovery and Development

2021 ◽  
pp. 247255522110309
Author(s):  
Olive Jung ◽  
Min Jae Song ◽  
Marc Ferrer
Keyword(s):  
Drug Discovery ◽  
Functional Characterization ◽  
Safety Testing ◽  
Drug Discovery And Development ◽  
Cellular Models ◽  
Clinical Biomarkers ◽  
Wide Range ◽  
Development Pipeline ◽  
Physiological Complexity

A wide range of complex in vitro models (CIVMs) are being developed for scientific research and preclinical drug efficacy and safety testing. The hope is that these CIVMs will mimic human physiology and pathology and predict clinical responses more accurately than the current cellular models. The integration of these CIVMs into the drug discovery and development pipeline requires rigorous scientific validation, including cellular, morphological, and functional characterization; benchmarking of clinical biomarkers; and operationalization as robust and reproducible screening platforms. It will be critical to establish the degree of physiological complexity that is needed in each CIVM to accurately reproduce native-like homeostasis and disease phenotypes, as well as clinical pharmacological responses. Choosing which CIVM to use at each stage of the drug discovery and development pipeline will be driven by a fit-for-purpose approach, based on the specific disease pathomechanism to model and screening throughput needed. Among the different CIVMs, biofabricated tissue equivalents are emerging as robust and versatile cellular assay platforms. Biofabrication technologies, including bioprinting approaches with hydrogels and biomaterials, have enabled the production of tissues with a range of physiological complexity and controlled spatial arrangements in multiwell plate platforms, which make them amenable for medium-throughput screening. However, operationalization of such 3D biofabricated models using existing automation screening platforms comes with a unique set of challenges. These challenges will be discussed in this perspective, including examples and thoughts coming from a laboratory dedicated to designing and developing assays for automated screening.

scholarly journals Genetically encoded ratiometric biosensor for probing lysosomal pH in mammalian cells and C. elegans

2020 ◽  
Author(s):  
Marcus Y. Chin ◽  
Anand R. Patwardhan ◽  
Kean-Hooi Ang ◽  
Austin L. Wang ◽  
Carolina Alquezar ◽  
...  
Keyword(s):  
Drug Discovery ◽  
Mammalian Cells ◽  
Fluorescent Protein ◽  
Multicellular Organism ◽  
Bafilomycin A1 ◽  
Atpase Inhibitor ◽  
High Content Imaging ◽  
Lysosomal Ph ◽  
C Elegans ◽  
Cellular Models

ABSTRACTLysosomes are important sites for macromolecular degradation, defined by a profoundly acidic lumenal pH of ~4.5. To better understand lysosomal pH, we designed a novel, genetically encoded, fluorescent protein-based pH biosensor called FIRE-pHLy (Fluorescence Indicator REporting pH in Lysosomes). This sensor was targeted to lysosomes with LAMP1 and reported lumenal pH between 3.0 and 6.0 with monomeric teal fluorescent protein 1 (mTFP1), a bright cyan FP variant with a pKa of 4.3. Ratiometric quantification was enabled with cytosolically oriented mCherry using high-content imaging. We expressed FIRE-pHLy in several cellular models as well as the multicellular organism C. elegans and quantified its alkalinizing response to bafilomycin A1, a specific V-ATPase inhibitor. In summary, we have engineered FIRE-pHLy, a specific, robust and versatile lysosomal pH biosensor that has broad applications for investigating pH dynamics in aging and lysosome-related diseases, as well as in lysosome-based drug discovery.

scholarly journals Alzheimer’s Risk Gene TREM2 Determines Functional Properties of New Type of Human iPSC-Derived Microglia

2021 ◽  
Vol 11 ◽  
Author(s):  
Marvin Reich ◽  
Iñaki Paris ◽  
Martin Ebeling ◽  
Nadine Dahm ◽  
Christophe Schweitzer ◽  
...  
Keyword(s):  
Drug Discovery ◽  
Large Scale ◽  
Expression Profiles ◽  
Well Being ◽  
Cytosolic Calcium ◽  
Regulatory Function ◽  
Mrna Levels ◽  
Cellular Models ◽  
Risk Gene ◽  
Human Ipsc

Microglia are key in the homeostatic well-being of the brain and microglial dysfunction has been implicated in neurodegenerative disorders such as Alzheimer’s disease (AD). Due to the many limitations to study microglia in situ or isolated for large scale drug discovery applications, there is a high need to develop robust and scalable human cellular models of microglia with reliable translatability to the disease. Here, we describe the generation of microglia-like cells from human induced pluripotent stem cells (iPSC) with distinct phenotypes for mechanistic studies in AD. We started out from an established differentiation protocol to generate primitive macrophage precursors mimicking the yolk sac ontogeny of microglia. Subsequently, we tested 36 differentiation conditions for the cells in monoculture where we exposed them to various combinations of media, morphogens, and extracellular matrices. The optimized protocol generated robustly ramified cells expressing key microglial markers. Bulk mRNA sequencing expression profiles revealed that compared to cells obtained in co-culture with neurons, microglia-like cells derived from a monoculture condition upregulate mRNA levels for Triggering Receptor Expressed On Myeloid Cells 2 (TREM2), which is reminiscent to the previously described disease-associated microglia. TREM2 is a risk gene for AD and an important regulator of microglia. The regulatory function of TREM2 in these cells was confirmed by comparing wild type with isogenic TREM2 knock-out iPSC microglia. The TREM2-deficient cells presented with stronger increase in free cytosolic calcium upon stimulation with ATP and ADP, as well as stronger migration towards complement C5a, compared to TREM2 expressing cells. The functional differences were associated with gene expression modulation of key regulators of microglia. In conclusion, we have established and validated a work stream to generate functional human iPSC-derived microglia-like cells by applying a directed and neuronal co-culture independent differentiation towards functional phenotypes in the context of AD. These cells can now be applied to study AD-related disease settings and to perform compound screening and testing for drug discovery.

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