High-throughput screening of a small molecule library for promoters and inhibitors of mesenchymal stem cell osteogenic differentiation

2010 ◽  
Vol 108 (1) ◽  
pp. 163-174 ◽  
Author(s):  
Darren M. Brey ◽  
Nuzhat A. Motlekar ◽  
Scott L. Diamond ◽  
Robert L. Mauck ◽  
Jonathon P. Garino ◽  
...  
Keyword(s):  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Osteogenic Differentiation ◽  
Small Molecule ◽  
High Throughput ◽  
High Throughput Screening ◽  
Small Molecule Library

High-Throughput Screening for Modulators of Mesenchymal Stem Cell Chondrogenesis

2008 ◽  
Vol 36 (11) ◽  
pp. 1909-1921 ◽  
Author(s):  
Alice H. Huang ◽  
Nuzhat A. Motlekar ◽  
Ashley Stein ◽  
Scott L. Diamond ◽  
Eileen M. Shore ◽  
...  
Keyword(s):  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
High Throughput ◽  
High Throughput Screening

scholarly journals High-Throughput Fluorescence-Based Activity Assay for Arginase-1

2020 ◽  
Vol 25 (9) ◽  
pp. 1018-1025
Author(s):  
Yvonne Grobben ◽  
Nicole Willemsen-Seegers ◽  
Joost C. M. Uitdehaag ◽  
Jos de Man ◽  
Jan van Groningen ◽  
...  
Keyword(s):  
Small Molecule ◽  
High Throughput ◽  
High Throughput Screening ◽  
Drug Target ◽  
Rank Order ◽  
Activity Assay ◽  
Arginase 1 ◽  
Cell Immunity ◽  
Small Molecule Library ◽  
Kinetics Of

Arginase-1, which converts the amino acid L-arginine into L-ornithine and urea, is a promising new drug target for cancer immunotherapy, as it has a role in the regulation of T-cell immunity in the tumor microenvironment. To enable the discovery of small-molecule Arginase-1 inhibitors by high-throughput screening, we developed a novel homogeneous (mix-and-measure) fluorescence-based activity assay. The assay measures the conversion of L-arginine into L-ornithine by a decrease in fluorescent signal due to quenching of a fluorescent probe, Arginase Gold. This way, inhibition of Arginase-1 results in a gain of signal when compared with the uninhibited enzyme. Side-by-side profiling of reference inhibitors in the fluorescence-based assay and a colorimetric urea formation assay revealed similar potencies and the same potency rank order among the two assay formats. The fluorescence-based assay was successfully automated for high-throughput screening of a small-molecule library in 384-well format with a good Z′-factor and hit confirmation rate. Finally, we show that the assay can be used to study the binding kinetics of inhibitors.

Exploring the mesenchymal stem cell niche using high throughput screening

Biomaterials ◽  
2013 ◽  
Vol 34 (31) ◽  
pp. 7601-7615 ◽  
Author(s):  
Soraya Rasi Ghaemi ◽  
Frances J. Harding ◽  
Bahman Delalat ◽  
Stan Gronthos ◽  
Nicolas H. Voelcker
Keyword(s):  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
High Throughput ◽  
High Throughput Screening ◽  
Stem Cell Niche ◽  
Cell Niche

High-Throughput Screening of Rat Mesenchymal Stem Cell Behavior on Gradient TiO2 Nanotubes

2018 ◽  
Vol 4 (8) ◽  
pp. 2804-2814 ◽  
Author(s):  
Ping Mu ◽  
Yanran Li ◽  
Yanmei Zhang ◽  
Yun Yang ◽  
Ren Hu ◽  
...  
Keyword(s):  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
High Throughput ◽  
High Throughput Screening ◽  
Tio2 Nanotubes ◽  
Cell Behavior

scholarly journals Development of a Novel High-Throughput Screen and Identification of Small-Molecule Inhibitors of the Gα–RGS17 Protein–Protein Interaction Using AlphaScreen

2011 ◽  
Vol 16 (8) ◽  
pp. 869-877 ◽  
Author(s):  
Duncan I. Mackie ◽  
David L. Roman
Keyword(s):  
Small Molecule ◽  
High Throughput ◽  
Protein Interaction ◽  
High Throughput Screening ◽  
Drug Targets ◽  
Screening Method ◽  
Fold Increase ◽  
Rgs Proteins ◽  
High Throughput Screen ◽  
Protein Protein Interaction

In this study, the authors used AlphaScreen technology to develop a high-throughput screening method for interrogating small-molecule libraries for inhibitors of the Gαo–RGS17 interaction. RGS17 is implicated in the growth, proliferation, metastasis, and the migration of prostate and lung cancers. RGS17 is upregulated in lung and prostate tumors up to a 13-fold increase over patient-matched normal tissues. Studies show RGS17 knockdown inhibits colony formation and decreases tumorigenesis in nude mice. The screen in this study uses a measurement of the Gαo–RGS17 protein–protein interaction, with an excellent Z score exceeding 0.73, a signal-to-noise ratio >70, and a screening time of 1100 compounds per hour. The authors screened the NCI Diversity Set II and determined 35 initial hits, of which 16 were confirmed after screening against controls. The 16 compounds exhibited IC50 <10 µM in dose–response experiments. Four exhibited IC50 values <6 µM while inhibiting the Gαo–RGS17 interaction >50% when compared to a biotinylated glutathione-S-transferase control. This report describes the first high-throughput screen for RGS17 inhibitors, as well as a novel paradigm adaptable to many other RGS proteins, which are emerging as attractive drug targets for modulating G-protein-coupled receptor signaling.

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