scholarly journals In vitro Assays and Imaging Methods for Drug Discovery for Cardiac Fibrosis

2021 ◽  
Vol 12 ◽  
Author(s):  
Giorgia Palano ◽  
Ariana Foinquinos ◽  
Erik Müllers
Keyword(s):  
Image Analysis ◽  
Drug Discovery ◽  
High Throughput Screening ◽  
Large Scale ◽  
Cardiac Fibrosis ◽  
3D Models ◽  
In Vitro Assays ◽  
Stress Injury

As a result of stress, injury, or aging, cardiac fibrosis is characterized by excessive deposition of extracellular matrix (ECM) components resulting in pathological remodeling, tissue stiffening, ventricular dilatation, and cardiac dysfunction that contribute to heart failure (HF) and eventually death. Currently, there are no effective therapies specifically targeting cardiac fibrosis, partially due to limited understanding of the pathological mechanisms and the lack of predictive in vitro models for high-throughput screening of antifibrotic compounds. The use of more relevant cell models, three-dimensional (3D) models, and coculture systems, together with high-content imaging (HCI) and machine learning (ML)-based image analysis, is expected to improve predictivity and throughput of in vitro models for cardiac fibrosis. In this review, we present an overview of available in vitro assays for cardiac fibrosis. We highlight the potential of more physiological 3D cardiac organoids and coculture systems and discuss HCI and automated artificial intelligence (AI)-based image analysis as key methods able to capture the complexity of cardiac fibrosis in vitro. As 3D and coculture models will soon be sufficiently mature for application in large-scale preclinical drug discovery, we expect the combination of more relevant models and high-content analysis to greatly increase translation from in vitro to in vivo models and facilitate the discovery of novel targets and drugs against cardiac fibrosis.

scholarly journals Recent Advances in In-Vitro Assays for Type 2 Diabetes Mellitus: An Overview

2020 ◽  
Vol 16 (1) ◽  
pp. 13-23
Author(s):  
Nazmina Vhora ◽  
Ujjal Naskar ◽  
Aishwarya Hiray ◽  
Abhijeet S. Kate ◽  
Alok Jain
Keyword(s):  
Type 2 Diabetes ◽  
Drug Discovery ◽  
High Throughput ◽  
High Throughput Screening ◽  
Antidiabetic Activity ◽  
Screening Methods ◽  
Selection Of

BACKGROUND: A higher rate of attenuation of molecules in drug discovery has enabled pharmaceutical companies to enhance the efficiency of their hit identification and lead optimization. Selection and development of appropriate in-vitro and in-vivo strategies may improve this process as primary and secondary screening utilize both strategies. In-vivo approaches are too relentless and expensive for assessing hits. Therefore, it has become indispensable to develop and implement suitable in-vitro screening methods to execute the required activities and meet the respective targets. However, the selection of an appropriate in-vitro assay for specific evaluation of cellular activity is no trivial task. It requires thorough investigation of the various parameters involved. AIM: In this review, we aim to discuss in-vitro assays for type 2 diabetes (T2D), which have been utilized extensively by researchers over the last five years, including target-based, non-target based, low-throughput, and high-throughput screening assays. METHODS: The literature search was conducted using databases including Scifinder, PubMed, ScienceDirect, and Google Scholar to find the significant published articles. DISCUSSION and CONCLUSION: The accuracy and relevance of in-vitro assays have a significant impact on the drug discovery process for T2D, especially in assessing the antidiabetic activity of compounds and identifying the site of effect in high-throughput screening. The report reviews the advantages, limitations, quality parameters, and applications of the probed invitro assays, and compares them with one another to enable the selection of the optimal method for any purpose. The information on these assays will accelerate numerous procedures in the drug development process with consistent quality and accuracy.

scholarly journals High Throughput Screening of Pharmacokinetics and Metabolism in Drug Discovery (II) —Investigation on In vitro and In vivo Correlation in Drug Metabolism Screening—

2005 ◽  
Vol 125 (1) ◽  
pp. 131-139 ◽  
Author(s):  
Hiroshi KOMURA ◽  
Kenichi MATSUDA ◽  
Yukie SHIGEMOTO ◽  
Iichiro KAWAHARA ◽  
Rieko ANO ◽  
...  
Keyword(s):  
Drug Discovery ◽  
Drug Metabolism ◽  
High Throughput ◽  
High Throughput Screening

scholarly journals A retrospective analysis to explore the applicability of fish biomarkers and sediment bioassays along contaminated salinity transects

2009 ◽  
Vol 66 (10) ◽  
pp. 2089-2105 ◽  
Author(s):  
Cor A. Schipper ◽  
Joost Lahr ◽  
Paul J. van den Brink ◽  
Steve G. George ◽  
Peter-Diedrich Hansen ◽  
...  
Keyword(s):  
Retrospective Analysis ◽  
Large Scale ◽  
Biological Effects ◽  
In Vitro Assays ◽  
Multivariate Statistical ◽  
Large Scale Field ◽  
Hepatic Metallothionein ◽  
Fish Biomarkers

Abstract Schipper, C. A., Lahr, J., van den Brink, P. J., George, S. G., Hansen, P-D., da Silva de Assis, H. C., van der Oost, R., Thain, J. E., Livingstone, D., Mitchelmore, C., van Schooten, F-J., Ariese, F., Murk, A. J., Grinwis, G. C. M., Klamer, H., Kater, B. J., Postma, J. F., van der Werf, B., and Vethaak, A. D. 2009. A retrospective analysis to explore the applicability of fish biomarkers and sediment bioassays along contaminated salinity transects. – ICES Journal of Marine Science, 66: 2089–2105. Biological-effects monitoring in estuarine environments is complex as a result of strong gradients and fluctuations in salinity and other environmental conditions, which may influence contaminant bioavailability and the physiology and metabolism of the organisms. To select the most robust and reliable biological-effect methods for monitoring and assessment programmes, a large-scale field study was conducted in two estuarine transects in the Netherlands. The locations ranged from heavily polluted harbour areas (the ports of Rotterdam and Amsterdam) to cleaner coastal and freshwater sites. Assessment methods used included a variety of biomarkers in flounder (Platichthys flesus) and a range of in vitro (sediment extracts) and in vivo bioassays. Multivariate statistical analysis was applied to investigate correlations and relationships between various biological effects and contaminant levels in flounder liver or sediments. Several biological methods seemed to be too much affected by salinity differences for routine use in estuaries. The most discriminative biomarkers in the study were hepatic metallothionein content and biliary 1-OH pyrene in fish. Mechanism-based in vitro assays DR-CALUX and ER-CALUX applied to sediment extracts for screening of potential toxicity were much more responsive than in vivo bioassays with macro-invertebrates using survival as an endpoint.

The Use of On-line and Off-line Chromatographic Extraction Techniques coupled with Mass Spectrometry for Support of In Vivo and In Vitro Assays in Drug Discovery

2001 ◽  
Vol 1 (5) ◽  
pp. 463-471 ◽  
Author(s):  
John A. Masucci ◽  
Gary W. Caldwell ◽  
William J. Jones ◽  
Stephen J. Juzwin ◽  
Patrick J. Sasso ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Drug Discovery ◽  
In Vitro Assays ◽  
Extraction Techniques ◽  
On Line ◽  
Chromatographic Extraction

scholarly journals 1341. Development of a Series of High-Throughput Screens to Identify Leads for Nontuberculous Mycobacteria Drug Design

2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S485-S485
Author(s):  
Sarah McGuffin ◽  
Steven Mullen ◽  
Julie Early ◽  
Tanya Parish
Keyword(s):  
Drug Discovery ◽  
Drug Development ◽  
High Throughput ◽  
Nontuberculous Mycobacteria ◽  
Human Infection ◽  
Attrition Rate ◽  
In Vitro Assays ◽  
Vitro Assay

Abstract Background Nontuberculous mycobacteria (NTM), particularly Mycobacterium avium complex and Mycobacterium abscessus complex, cause significant morbidity and mortality in patients with impaired host immunity or pre-existing structural lung conditions. NTM infections are increasing at an alarming rate worldwide and there is a dearth of progress in regard to the development of efficacious and tolerable drugs to treat such infections. Traditional drug discovery screens do not account for the diverse physiological conditions, microenvironments, and compartments that the bacilli encounter during human infection. In order to help populate the NTM drug pipeline, and explore the disconnect between in vitro activity, in vivo activity, and clinical outcomes, we are developing a high throughput in vitro assay platform that will more closely model the unique infection-relevant conditions encountered by NTM. Methods We are developing and validating a suite of in vitro assays that screen compounds for activity against extracellular planktonic bacteria, extracellular bacteria within biofilms, intracellular bacteria, and nutrient-starved non-replicating bacteria. Results We are using both the smooth and rough morphotypes of M. abscessus and M. avium. We have validated high throughput assays to pharmaceutical standards for replicating and non-replicating M. abscessus. We have also tested a panel of 18 known anti-mycobacterial compounds. Assay development is currently underway to test compounds for activity against NTM in biofilm and inside macrophages as well. Conclusion To enhance hit identification for scaffolds to use as starting points for NTM drug development, focused libraries of compounds that have undergone significant preclinical profiling and/or compounds with known activity against M. tuberculosis (TB) will be screened. Such a “piggyback” approach usurps advances made in TB drug development and leverages them for NTM drug discovery. This will help expedite novel drug development, reduce attrition rate, and offer a shorter route to clinical use as it exploits the prior investment in medicinal chemistry, pharmacology, and toxicology. Disclosures All authors: No reported disclosures.

scholarly journals Mechanism-Driven Read-Across of Chemical Hepatotoxicants Based on Chemical Structures and Biological Data

2020 ◽  
Vol 174 (2) ◽  
pp. 178-188 ◽  
Author(s):  
Linlin Zhao ◽  
Daniel P Russo ◽  
Wenyi Wang ◽  
Lauren M Aleksunes ◽  
Hao Zhu
Keyword(s):  
High Throughput Screening ◽  
Drug Efficacy ◽  
Biological Data ◽  
In Vitro Assays ◽  
In Vitro Toxicity ◽  
Chemical Toxicity ◽  
Chemical Structures ◽  
Modeling Strategy

Abstract Hepatotoxicity is a leading cause of attrition in the drug development process. Traditional preclinical and clinical studies to evaluate hepatotoxicity liabilities are expensive and time consuming. With the advent of critical advancements in high-throughput screening, there has been a rapid accumulation of in vitro toxicity data available to inform the risk assessment of new pharmaceuticals and chemicals. To this end, we curated and merged all available in vivo hepatotoxicity data obtained from the literature and public resources, which yielded a comprehensive database of 4089 compounds that includes hepatotoxicity classifications. After dividing the original database of chemicals into modeling and test sets, PubChem assay data were automatically extracted using an in-house data mining tool and clustered based on relationships between structural fragments and cellular responses in in vitro assays. The resultant PubChem assay clusters were further investigated. During the cross-validation procedure, the biological data obtained from several assay clusters exhibited high predictivity of hepatotoxicity and these assays were selected to evaluate the test set compounds. The read-across results indicated that if a new compound contained specific identified chemical fragments (ie, Molecular Initiating Event) and showed active responses in the relevant selected PubChem assays, there was potential for the chemical to be hepatotoxic in vivo. Furthermore, several mechanisms that might contribute to toxicity were derived from the modeling results including alterations in nuclear receptor signaling and inhibition of DNA repair. This modeling strategy can be further applied to the investigation of other complex chemical toxicity phenomena (eg, developmental and reproductive toxicities) as well as drug efficacy.

scholarly journals Rapid Microbiologic and Pharmacologic Evaluation of Experimental Compounds against Mycobacterium tuberculosis

2006 ◽  
Vol 50 (4) ◽  
pp. 1245-1250 ◽  
Author(s):  
Veronica Gruppo ◽  
Christine M. Johnson ◽  
Karen S. Marietta ◽  
Hataichanok Scherman ◽  
Erin E. Zink ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Drug Discovery ◽  
High Throughput Screening ◽  
Mouse Serum ◽  
Serum Samples ◽  
Microtiter Plate Assay ◽  
Plate Assay ◽  
Drug Concentrations

ABSTRACT The assessment of physiochemical and pharmacological properties at early stages of drug discovery can accelerate the conversion of hits and leads into candidates for further development. A strategy for streamlined evaluation of compounds against Mycobacterium tuberculosis in the early preclinical stage is presented in this report. As a primary assay to rapidly select experimental compounds with sufficient in vitro activity, the growth inhibition microtiter plate assay was devised as an alternative to current methods. This microdilution plate assay is a liquid culture method based on spectrophotometric readings of the bacillary growth. The performance of this method was compared to the performance of two established susceptibility methods using clinical available tuberculosis (TB) drugs. Data generated from all three assays were similar for all of the tested compounds. A second simple bioassay was devised to assess the oral bioavailability of compounds prior to extensive in vivo efficacy testing. The bioassay estimates drug concentrations in collected serum samples by a microdilution MIC plate method using M. tuberculosis. In the same assay, the MIC of the compound is also determined in the presence of 10% mouse serum as an indication of protein binding. The method was validated using different clinically available TB drugs, and results are discussed in this report. With these methodological advances, screening of compounds against tuberculosis in the preclinical phase will be rapid, can be adapted to semi-high-throughput screening, and will add relevant physicochemical and basic pharmacological criteria to the decision process of drug discovery.

scholarly journals Development of a miniaturized 3D organoid culture platform for ultra-high-throughput screening

2020 ◽  
Vol 12 (8) ◽  
pp. 630-643 ◽  
Author(s):  
Yuhong Du ◽  
Xingnan Li ◽  
Qiankun Niu ◽  
Xiulei Mo ◽  
Min Qui ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Drug Discovery ◽  
High Throughput ◽  
High Throughput Screening ◽  
Large Scale ◽  
Genetically Engineered ◽  
High Density ◽  
Organoid Culture ◽  
3D Architecture

Abstract The recent advent of robust methods to grow human tissues as 3D organoids allows us to recapitulate the 3D architecture of tumors in an in vitro setting and offers a new orthogonal approach for drug discovery. However, organoid culturing with extracellular matrix to support 3D architecture has been challenging for high-throughput screening (HTS)-based drug discovery due to technical difficulties. Using genetically engineered human colon organoids as a model system, here we report our effort to miniaturize such 3D organoid culture with extracellular matrix support in high-density plates to enable HTS. We first established organoid culturing in a 384-well plate format and validated its application in a cell viability HTS assay by screening a 2036-compound library. We further miniaturized the 3D organoid culturing in a 1536-well ultra-HTS format and demonstrated its robust performance for large-scale primary compound screening. Our miniaturized organoid culturing method may be adapted to other types of organoids. By leveraging the power of 3D organoid culture in a high-density plate format, we provide a physiologically relevant screening platform to model tumors to accelerate organoid-based research and drug discovery.

scholarly journals 3D Cell-Based Assays for Drug Screens: Challenges in Imaging, Image Analysis, and High-Content Analysis

2019 ◽  
Vol 24 (6) ◽  
pp. 615-627 ◽  
Author(s):  
Tijmen H. Booij ◽  
Leo S. Price ◽  
Erik H. J. Danen
Keyword(s):  
Content Analysis ◽  
Drug Discovery ◽  
High Throughput Screening ◽  
3D Cell Culture ◽  
Automated Analysis ◽  
In Vivo Models ◽  
Current Switch ◽  
High Content Analysis

The introduction of more relevant cell models in early preclinical drug discovery, combined with high-content imaging and automated analysis, is expected to increase the quality of compounds progressing to preclinical stages in the drug development pipeline. In this review we discuss the current switch to more relevant 3D cell culture models and associated challenges for high-throughput screening and high-content analysis. We propose that overcoming these challenges will enable front-loading the drug discovery pipeline with better biology, extracting the most from that biology, and, in general, improving translation between in vitro and in vivo models. This is expected to reduce the proportion of compounds that fail in vivo testing due to a lack of efficacy or to toxicity.

scholarly journals Predictability of Peripheral Lymphocyte Reduction of Novel S1P1 Agonists by In Vitro GPCR Signaling Profile

2013 ◽  
Vol 18 (9) ◽  
pp. 997-1007 ◽  
Author(s):  
Han Xu ◽  
Michele McElvain ◽  
Mike Fiorino ◽  
Brad Henkle ◽  
Lisa Sherman ◽  
...  
Keyword(s):  
Drug Discovery ◽  
Adenylyl Cyclase ◽  
Peripheral Lymphocyte ◽  
Receptor Internalization ◽  
In Vitro Assays ◽  
Vitro Assay ◽  
In Vivo Efficacy ◽  
Erk Phosphorylation

Surrogate readouts of G-protein–coupled receptor signaling pathways using highly engineered systems are often employed in the drug discovery process. However, accumulating data have demonstrated the importance of selecting relevant biological activity rather than technically facile assays to support high-throughout screening and subsequent structure-activity relationship studies. Here we report a case study using sphingosine-1-phosphate receptor 1 (S1P1) as the model system to compare compound activity in six different in vitro assays with their ability to predict in vivo efficacy. S1P1 has long been validated as a therapeutic target for autoimmune diseases. In this article, in vivo and in vitro studies on 19 S1P1 agonists are reported. In vitro activities of these S1P1 agonists, together with S1P and FTY720p, on Ca2+ mobilization, adenylyl cyclase inhibition, extracellular signal-related kinase (ERK) phosphorylation, β-arrestin recruitment, and receptor internalization, were determined. The in vitro potency of these compounds was correlated with their ability to induce peripheral lymphocyte reduction. The results revealed that inhibition of adenylyl cyclase and induction of β-arrestin recruitment and receptor internalization are good indicators to predict in vivo efficacy, whereas induction of Ca2+ mobilization through Gqi/5 coupling and ERK phosphorylation is irrelevant. This study demonstrated the importance of identifying an appropriate in vitro assay to predict in vivo activity based on the biological relevance in the drug discovery setting.

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