High-throughput screening to identify inhibitors of lysine demethylases

Gale, Molly; Yan, Qin

doi:10.2217/epi.14.63

Cited by 19 publications

(31 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We performed a screen using the full-length KDM5A protein, an approach which enables identification of new inhibitor chemotypes that may not be discovered by screening against truncated KDM5A or by structure-guided design. The screen was performed using the AlphaScreen platform, which is cost-effective, highly sensitive, and requires only small amounts of enzyme (for review of demethylase assays see [32]).…”

Section: Discussionmentioning

confidence: 99%

Screen-identified selective inhibitor of lysine demethylase 5A blocks cancer cell growth and drug resistance

et al. 2016

Self Cite

View full text Add to dashboard Cite

Lysine demethylase 5A (KDM5A/RBP2/JARID1A) is a histone lysine demethylase that is overexpressed in several human cancers including lung, gastric, breast and liver cancers. It plays key roles in important cancer processes including tumorigenesis, metastasis, and drug tolerance, making it a potential cancer therapeutic target. Chemical tools to analyze KDM5A demethylase activity are extremely limited as available inhibitors are not specific for KDM5A. Here, we characterized KDM5A using a homogeneous luminescence-based assay and conducted a screen of about 9,000 small molecules for inhibitors. From this screen, we identified several 3-thio-1,2,4-triazole compounds that inhibited KDM5A with low μM in vitro IC50 values. Importantly, these compounds showed great specificity and did not inhibit its close homologue KDM5B (PLU1/JARID1B) or the related H3K27 demethylases KDM6A (UTX) and KDM6B (JMJD3). One compound, named YUKA1, was able to increase H3K4me3 levels in human cells and selectively inhibit the proliferation of cancer cells whose growth depends on KDM5A. As KDM5A was shown to mediate drug tolerance, we investigated the ability of YUKA1 to prevent drug tolerance in EGFR-mutant lung cancer cells treated with gefitinib and HER2+ breast cancer cells treated with trastuzumab. Remarkably, this compound hindered the emergence of drug-tolerant cells, highlighting the critical role of KDM5A demethylase activity in drug resistance. The small molecules presented here are excellent tool compounds for further study of KDM5A's demethylase activity and its contributions to cancer.

show abstract

Section: Discussionmentioning

confidence: 99%

Screen-identified selective inhibitor of lysine demethylase 5A blocks cancer cell growth and drug resistance

et al. 2016

Self Cite

View full text Add to dashboard Cite

show abstract

“…Broad range HDACi, including TSA and SAHA, have already been used as potential pharmaceutical targets in arthritis, colitis and SLE exhibiting promising results (Glauben et al, ; Lin et al, ; Mishra, Reilly, Brown, Ruiz, & Gilkeson, ). In the past few years, various KDM inhibitors have been identified (Gale and Yan, ; Kruidenier et al, ; Thinnes et al, ). Several of these selectively target the H3K27me3‐demethylating KDM6 enzymes and attenuate the inflammatory response in activated macrophages, encouraging the design and use of KDM inhibitors in inflammatory disease treatment (Kruidenier et al, ).…”

Section: Contribution Of Histone Modifying Enzymes In Inflammatory DImentioning

confidence: 99%

Histone methylation and acetylation in macrophages as a mechanism for regulation of inflammatory responses

Daskalaki

Tsatsanis

Kampranis

2018

Journal Cellular Physiology

100

View full text Add to dashboard Cite

Macrophages respond to noxious stimuli and contribute to inflammatory responses by eliminating pathogens or damaged tissue and maintaining homeostasis. Response to activation signals and maintenance of homeostasis require tight regulation of genes involved in macrophage activation and inactivation processes, as well as genes involved in determining their polarization state. Recent evidence has revealed that such regulation occurs through histone modifications that render inflammatory or polarizing gene promoters accessible to transcriptional complexes. Thus, inflammatory and anti-inflammatory genes are regulated by histone acetylation and methylation, determining their activation state. Herein, we review the current knowledge on the role of histone modifying enzymes (acetyltransferases, deacetylases, methyltransferases, and demethylases) in determining the responsiveness and M1 or M2 polarization of macrophages. The contribution of these enzymes in the development of inflammatory diseases is also presented.

show abstract

“…An advantage of this assay is the use of the biotinylated peptide, which allows for washout of all other assay components before treatment with antibody. A related assay which uses the europium antibody is called LANCE Ultra where FRET occurs between the donor europium-labeled primary antibody to the site of interest and the acceptor ULight (Gale&Yan, 2015). Here, the peptide is N-terminally bound by primary antibody and FRET occurs between the europium and ULight at the C-terminal end.…”

Section: Lsd1 Assaysmentioning

confidence: 99%

“…Here, the peptide is N-terminally bound by primary antibody and FRET occurs between the europium and ULight at the C-terminal end. An Alpha Screen that is analogous to LANCE Ultra has also been reported (Gale&Yan, 2015). Library screening can be pursued with the DEL-FIA assay since no fluorescence or quenching of the compounds is likely to occur.…”

Section: Lsd1 Assaysmentioning

confidence: 99%

“…SAMDI has been used in a high-throughput screen to identify LSD1 inhibitors. Another mass spectrometry-based assay to analyze demethylation is RapidFire that uses tagged peptides, which are then pooled (Gale & Yan, 2015). Mass spectrometry-based methods are sensitive, can be made label free, and require very little amounts of enzyme and substrate.…”

Section: Lsd1 Assaysmentioning

confidence: 99%

See 1 more Smart Citation

LSD1 Histone Demethylase Assays and Inhibition

Hayward¹,

Cole²

2016

Methods in Enzymology

View full text Add to dashboard Cite

The lysine-specific demethylase (LSD1) is a flavin-dependent amine oxidase that selectively removes one or two methyl groups from histone H3 at the Lys4 position. Along with histone deacetylases 1 and 2, LSD1 is involved in epigenetically silencing gene expression. LSD1 has been implicated as a potential therapeutic target in cancer and other diseases. In this chapter, we discuss several approaches to measure LSD1 demethylase activity and their relative strengths and limitations for inhibitor discovery and mechanistic characterization. In addition, we review the principal established chemical functional groups derived from monoamine oxidase inhibitors that have been investigated in the context of LSD1 as demethylase inhibitors. Finally, we highlight a few examples of recently developed LSD1 mechanism-based inactivators and their biomedical applications.

show abstract

High-throughput screening to identify inhibitors of lysine demethylases

Cited by 19 publications

References 47 publications

Screen-identified selective inhibitor of lysine demethylase 5A blocks cancer cell growth and drug resistance

Screen-identified selective inhibitor of lysine demethylase 5A blocks cancer cell growth and drug resistance

Histone methylation and acetylation in macrophages as a mechanism for regulation of inflammatory responses

LSD1 Histone Demethylase Assays and Inhibition

Contact Info

Product

Resources

About