scholarly journals Drug Efficacy Comparison of 3D Forming and Preforming Sphere Models with a Micropillar and Microwell Chip Platform

2019 ◽  
Vol 24 (4) ◽  
pp. 476-483 ◽  
Author(s):  
Il Doh ◽  
Yong-Jun Kwon ◽  
Bosung Ku ◽  
Dong Woo Lee
Keyword(s):  
High Throughput Screening ◽  
Drug Efficacy ◽  
High Sensitivity ◽  
Chemotherapeutic Agents ◽  
Cell Culture Model ◽  
Drug Candidates ◽  
Tumor Microenvironments ◽  
Culture Model

Hepatocellular carcinoma (HCC), a major histological subtype of liver cancer, is the third most common cause of cancer-related death worldwide. Currently, many curative standard treatments using target-specific chemotherapeutic agents are being developed. However, drug efficacy tests based on the 2D monolayer cell culture model do not effectively screen the best drug candidates because they do not accurately reflect in vivo tumor microenvironments. Thus, to select the best drug candidates or repositioning drugs, we developed new 3D in vitro hepatic tumor models, including 3D forming and preformed sphere models. A micropillar and microwell chip platform was used for the 3D in vitro liver cell-based model for high-throughput screening. We measured the efficacy of 60 drugs and sorted the most efficacious drugs by comparing the drug response of the 2D monolayer model with the 3D forming and preformed sphere models. Among the 60 drugs, 17 drugs (28.3%) showed a significant high efficacy in the 3D preformed sphere model, while 45 drugs (75%) showed an efficacy in the 2D model. We also calculated the IC50 values of the 17 drugs and found that 7 drugs exhibited a high sensitivity in HCC, which was in agreement with previous studies.

scholarly journals Ratiometric bioluminescent sensors towards in vivo imaging of bacterial signaling

2019 ◽  
Author(s):  
A. B Dippel ◽  
W. A. Anderson ◽  
J. H. Park ◽  
F. H. Yildiz ◽  
M.C. Hammond
Keyword(s):  
High Throughput Screening ◽  
High Sensitivity ◽  
Second Messenger ◽  
Live Cells ◽  
Diagnostic Tools ◽  
Signaling Networks ◽  
Mammalian Host ◽  
Host Infection

ABSTRACTSecond messenger signaling networks allow cells to sense and adapt to changing environmental conditions. In bacteria, the nearly ubiquitous second messenger molecule cyclic di-GMP coordinates diverse processes such as motility, biofilm formation, and virulence. In bacterial pathogens, these signaling networks allow the bacteria to survive changing environment conditions that are experienced during infection of a mammalian host. While studies have examined the effects of cyclic di-GMP levels on virulence in these pathogens, it has previously not been possible to visualize cyclic di-GMP levels in real time during the stages of host infection. Towards this goal, we generate the first ratiometric, chemiluminescent biosensor scaffold that selectively responds to c-di-GMP. By engineering the biosensor scaffold, a suite of Venus-YcgR-NLuc (VYN) biosensors is generated that provide extremely high sensitivity (KD < 300 pM) and large BRET signal changes (up to 109%). As a proof-of-concept that VYN biosensors can image cyclic di-GMP during host infection, we show that the VYN biosensors function in the context of a tissue phantom model, with only ∼103-104 biosensor-expressing cells required for the measurement. Furthermore, the stable BRET signal suggests that the sensors could be used for long-term imaging of cyclic di-GMP dynamics during host infection. The VYN sensors developed here can serve as robust in vitro diagnostic tools for high throughput screening, as well as genetically encodable tools for monitoring the dynamics of c-di-GMP in live cells, and lay the groundwork for live cell imaging of c-di-GMP dynamics in bacteria during host infection, and other complex environments.

scholarly journals Extensive transcriptional and chromatin changes underlie astrocyte maturation in vivo and in culture

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Michael Lattke ◽  
Robert Goldstone ◽  
James K. Ellis ◽  
Stefan Boeing ◽  
Jerónimo Jurado-Arjona ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Cell Culture Model ◽  
Functional Maturation ◽  
Culture Model ◽  
Mature Astrocyte ◽  
Transcriptional Changes ◽  
A Cell ◽  
Brain Homeostasis

AbstractAstrocytes have essential functions in brain homeostasis that are established late in differentiation, but the mechanisms underlying the functional maturation of astrocytes are not well understood. Here we identify extensive transcriptional changes that occur during murine astrocyte maturation in vivo that are accompanied by chromatin remodelling at enhancer elements. Investigating astrocyte maturation in a cell culture model revealed that in vitro-differentiated astrocytes lack expression of many mature astrocyte-specific genes, including genes for the transcription factors Rorb, Dbx2, Lhx2 and Fezf2. Forced expression of these factors in vitro induces distinct sets of mature astrocyte-specific transcripts. Culturing astrocytes in a three-dimensional matrix containing FGF2 induces expression of Rorb, Dbx2 and Lhx2 and improves astrocyte maturity based on transcriptional and chromatin profiles. Therefore, extrinsic signals orchestrate the expression of multiple intrinsic regulators, which in turn induce in a modular manner the transcriptional and chromatin changes underlying astrocyte maturation.

scholarly journals Dual Effects of NV-CoV-2 Biomimetic Polymer: An Antiviral regimen against COVID-19

2021 ◽  
Author(s):  
Ashok Chakraborty ◽  
Anil Diwan ◽  
Vijetha Chiniga ◽  
Vinod Arora ◽  
Preetam Holkar ◽  
...  
Keyword(s):  
Cell Culture ◽  
Model Experiment ◽  
Antiviral Drug ◽  
Cell Culture Model ◽  
Culture Model ◽  
Platform Technology ◽  
Biomimetic Polymer ◽  
The Stability

Remdesivir (RDV) is the only antiviral drug so far approved for COVID-19 therapy by the FDA. However its efficacy is limited in vivo due to its low stability in presence of plasma. This paper compared the stability of RDV encapsulated with our platform technology based polymer NV-387 (NV-CoV-2), in presence of plasma in vitro and in vivo . Furthermore, a non-clinical pharmacology studies of NV-CoV-2 (Polymer) and NV-CoV-2-R (Polymer encapsulated Remdesivir ) in both NL-63 infected and uninfected rats were done. In an in vitro cell culture model experiment, antiviral activity of NV-CoV-2 and NV-CoV-2-R are also compared with RDV.

scholarly journals Extensive transcriptional and chromatin changes underlie astrocyte maturation in vivo and in culture

2020 ◽  
Author(s):  
Michael Lattke ◽  
Robert Goldstone ◽  
Francois Guillemot
Keyword(s):  
Three Dimensional ◽  
Cell Culture Model ◽  
Culture Model ◽  
Mature Astrocyte ◽  
Transcriptional Changes ◽  
A Cell ◽  
Late Stages ◽  
Brain Homeostasis

SummaryAstrocytes have diverse functions in brain homeostasis. Many of these functions are acquired during late stages of differentiation when astrocytes become fully mature. The mechanisms underlying astrocyte maturation are not well understood. Here we identified extensive transcriptional changes that occur during astrocyte maturation and are accompanied by chromatin remodelling at enhancer elements. Investigating astrocyte maturation in a cell culture model revealed that in vitro-differentiated astrocytes lacked expression of many mature astrocyte-specific genes, including genes for the transcription factors Rorb, Dbx2, Lhx2 and Fezf2. Forced expression of these factors in vitro induced distinct sets of mature astrocytes-specific transcripts. Culturing astrocytes with FGF2 in a three-dimensional gel induced expression of Rorb, Dbx2 and Lhx2 and improved their maturity based on transcriptional and chromatin profiles. Therefore extrinsic signals orchestrate the expression of multiple intrinsic regulators, which in turn induce in a modular manner the transcriptional and chromatin changes underlying astrocyte maturation.

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 Discovery of Novel Antigiardiasis Drug Candidates

2014 ◽  
Vol 58 (12) ◽  
pp. 7303-7311 ◽  
Author(s):  
Liudmila Kulakova ◽  
Andrey Galkin ◽  
Catherine Z. Chen ◽  
Noel Southall ◽  
Juan J. Marugan ◽  
...  
Keyword(s):  
High Throughput Screening ◽  
Standard Care ◽  
Parasitic Infection ◽  
The United States ◽  
The European Union ◽  
Current Standard ◽  
Content Type ◽  
Drug Candidates

ABSTRACTGiardiasis is a severe intestinal parasitic disease caused byGiardia lamblia, which inflicts many people in poor regions and is the most common parasitic infection in the United States. Current standard care drugs are associated with undesirable side effects, treatment failures, and an increasing incidence of drug resistance. As follow-up to a high-throughput screening of an approved drug library, which identified compounds lethal toG. lambliatrophozoites, we have determined the minimum lethal concentrations of 28 drugs and advanced 10 of them toin vivostudies in mice. The results were compared to treatment with the standard care drug, metronidazole, in order to identify drugs with equal or better anti-Giardiaactivities. Three drugs, fumagillin, carbadox, and tioxidazole, were identified. These compounds were also potent against metronidazole-resistant humanG. lambliaisolates (assemblages A and B), as determined inin vitroassays. Of these three compounds, fumagillin is currently an orphan drug used within the European Union to treat microsporidiosis in immunocompromised individuals, whereas carbadox and tioxidazole are used in veterinary medicine. A dose-dependent study of fumagillin in a giardiasis mouse model revealed that the effective dose of fumagillin was ∼100-fold lower than the metronidazole dose. Therefore, fumagillin may be advanced to further studies as an alternative treatment for giardiasis when metronidazole fails.

scholarly journals Activities of roxithromycin against Mycobacterium avium infections in human macrophages and C57BL/6 mice.

1995 ◽  
Vol 39 (4) ◽  
pp. 878-881 ◽  
Author(s):  
L Struillou ◽  
Y Cohen ◽  
N Lounis ◽  
G Bertrand ◽  
J Grosset ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Human Monocyte ◽  
Cell Culture Model ◽  
Human Macrophages ◽  
End Of Treatment ◽  
Culture Model ◽  
Intracellular Activities ◽  
Established Infection

The activity of roxithromycin against three clinical isolates of Mycobacterium avium was compared with that of clarithromycin both in a model of infection of human monocyte-derived macrophages and in a model of established infection of C57BL/6 mice. In the cell culture model, roxithromycin and clarithromycin were bactericidal for strains MO-1 and N-92159 and bacteriostatic for strain N-93043. For the three strains, the differences between the intracellular activities of roxithromycin and clarithromycin were not singificant after 7 days of treatment. Mice were infected with the MO-1 strain. Drugs were given by gavage at a dosage of 200 mg/kg of body weight 6 days per week for 16 weeks starting 5 weeks after infection. At the end of treatment, clarithromycin was more effective than roxithromycin in lungs; roxithromycin was as effective as clarithromycin in spleens. Thus, the activity of roxithromycin was comparable to that of clarithromycin both in vitro and in vivo.

scholarly journals Unbiased transcriptomic analysis of chondrocyte differentiation in a high-density cell culture model

2021 ◽  
Author(s):  
Claudia Kruger ◽  
Aimee Limpach ◽  
Claudia Kappen
Keyword(s):  
Cell Culture ◽  
High Density ◽  
Cell Culture Model ◽  
Cartilage Differentiation ◽  
Culture Model ◽  
High Density Cell ◽  
Vertebrate Skeleton ◽  
High Density Cell Culture

ABSTRACTIn the developing vertebrate skeleton, cartilage is an important precursor to the formation of bones. Cartilage is produced by chondrocytes, which derive from embryonic mesoderm and undergo a stereotypical program of differentiation and maturation. Here we modeled this process in vitro, using primary fetal mouse rib chondrocytes in a high-density cell culture model of cartilage differentiation, and performed genome-wide gene expression profiling over the course of culture. The overarching goal of this study was to characterize the molecular pathways involved in cartilage differentiation and maturation. Our results also enable a comprehensive appraisal of distinctions between common in vitro models for cartilage differentiation, and of differences in their molecular resemblance to cartilage formation in vivo.

scholarly journals Phytochemicals and Potential Therapeutic Targets on Toxoplasma gondii Parasite

2020 ◽  
Vol 20 (9) ◽  
pp. 739-753
Author(s):  
Sharif Alhassan Abdullahi ◽  
Ngah Zasmy Unyah ◽  
Noshariza Nordin ◽  
Rusliza Basir ◽  
Wana Mohammed Nasir ◽  
...  
Keyword(s):  
Drug Target ◽  
Relevant Literature ◽  
Chemotherapeutic Agents ◽  
Potential Drug ◽  
Natural Sources ◽  
Drug Candidates ◽  
Target Sites ◽  
Phytochemical Compounds

Identification of drug target in protozoan T. gondii is an important step in the development of chemotherapeutic agents. Likewise, exploring phytochemical compounds effective against the parasite can lead to the development of new drug agent that can be useful for prophylaxis and treatment of toxoplasmosis. In this review, we searched for the relevant literature on the herbs that were tested against T. gondii either in vitro or in vivo, as well as different phytochemicals and their potential activities on T. gondii. Potential activities of major phytochemicals, such as alkaloid, flavonoid, terpenoids and tannins on various target sites on T. gondii as well as other related parasites was discussed. It is believed that the phytochemicals from natural sources are potential drug candidates for the treatment of toxoplasmosis with little or no toxicity to humans.

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