17-DMAG dually inhibits Hsp90 and histone lysine demethylases in alveolar rhabdomyosarcoma

Singh, Shivendra V.; Abu‐Zaid, Ahmed; Lin, Wenwei; Low, Jonathan; Abdolvahabi, Alireza; Jin, Hong; Wu, Qiong; Cooke, Bailey; Fang, Jie; Bowling, John J.; Vaithiyalingam, Sivaraja; Currier, Duane; Yun, Mi‐Kyung; Fernando, Dinesh M.; Maier, Julie; Tillman, Heather; Bulsara, Purva; Lu, Zhaohua; Das, Sourav; Shelat, Anang A.; Li, Zhenmei; Young, Brandon; Lee, Richard; Rankovic, Zoran; Murphy, Andrew J.; White, Stephen W.; Davidoff, Andrew M.; Chen, Taosheng; Yang, Jun

doi:10.1016/j.isci.2020.101996

Cited by 10 publications

(24 citation statements)

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“… To identify an optimal assay buffer, three buffers (buffer 1 to 3) are tested. The presence of KCl and MgCl 2 could negatively impact the KDM4B enzymatic activity, whereas the presence of Tris-HCl (pH 8.0) could positively increase KDM4B enzymatic activity (Supplementary Figure 1E and 1F in ( Singh et al., 2021 )). These observations indicate that salt and pH have significant impact on KDM4B activity in TR-FRET assay, and the mechanisms need to be further investigated.…”

Section: Step-by-step Methods Detailsmentioning

confidence: 99%

“… Note: This test is to demonstrate the specificity of Tb-anti-H3K9Me2 antibody to the H3K9Me2-Biotin, but not others. (Supplementary Figure 1A in ( Singh et al., 2021 )) In a 384-well black low volume assay plate, 2 nM Tb-anti-H3K9Me2 and 20 nM AF488-Streptavidin at 20 μL/well are incubated with 1:2 dilutions of the substrate peptide H3K9Me3-Biotin or the resulting product peptide H3K9Me2-Biotin for 30 min. Detection of the TR-FRET signals of each group.…”

Section: Step-by-step Methods Detailsmentioning

confidence: 99%

“… Note: This test is to demonstrate the specificity of Tb-anti-H3K9Me2 antibody to the resulting product peptide H3K9Me2-Biotin at all tested concentrations but not to the substrate peptide H3K9Me3-Biotin. (Supplementary Figure 1B in ( Singh et al., 2021 )) Note: Specific detection of the newly generated H3K9Me2 peptide by the Tb-anti-H3K9Me2 antibody confers the assay with great specificity. To identify an optimal assay buffer, three buffers (buffer 1 to 3) are tested.…”

Section: Step-by-step Methods Detailsmentioning

confidence: 99%

“…We describe this protocol for screen of KDM4B inhibitors, which can be modified to screen inhibitors of other JmjC-domain-containing KDMs. For complete details on the use and execution of this protocol, please refer to Singh et al. (2021) .…”

mentioning

confidence: 99%

“…For complete details on the use and execution of this protocol, please refer to Singh et al. (2021) .…”

mentioning

confidence: 99%

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A protocol for high-throughput screening of histone lysine demethylase 4 inhibitors using TR-FRET assay

Lin

et al. 2021

STAR Protocols

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Summary Identification of diverse chemotypes of selective KDM4 inhibitors is important for exploring and validating the roles of KDM4s in the pathogenesis of human disease and for developing therapies. Here, we report a protocol for high-throughput screening of KDM4 inhibitors using TR-FRET demethylation functional assay. We describe this protocol for screen of KDM4B inhibitors, which can be modified to screen inhibitors of other JmjC-domain-containing KDMs. For complete details on the use and execution of this protocol, please refer to Singh et al. (2021) .

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Section: Step-by-step Methods Detailsmentioning

confidence: 99%

Section: Step-by-step Methods Detailsmentioning

confidence: 99%

Section: Step-by-step Methods Detailsmentioning

confidence: 99%

mentioning

confidence: 99%

“…For complete details on the use and execution of this protocol, please refer to Singh et al. (2021) .…”

mentioning

confidence: 99%

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A protocol for high-throughput screening of histone lysine demethylase 4 inhibitors using TR-FRET assay

Lin

et al. 2021

STAR Protocols

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The Application of High‐Throughput Approaches in Identifying Novel Therapeutic Targets and Agents to Treat Diabetes

Lim

2022

Advanced Biology

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An early example of HTS in drug development came from the identification of new antibiotics using 96-well microtiter plates, whereby fermentations of an established actinomycetes library were incubated with a panel of microorganisms, and after optimization, a screening capacity of 10 000 fermentation broths per week was achieved. [1] In the following decades, the rapid development of molecular biology techniques and advances in automated equipment have changed how new drugs are identified, and a variety of HTS-identified drugs have been approved to treat different diseases. [2] For a comprehensive list of HTS-identified drugs, please refer to the excellent review by Macarron et al. [2] High-throughput applications can also be combined with other unbiased "Omics" technologies used to study genomics, epigenomics, transcriptomics, proteomics, and metabolomics. [3] Diabetes is an incurable disease characterized by the inability of the body to maintain normoglycemia due to decreased sensitivity of various tissues to insulin and insufficient amounts of circulating insulin, a hormone produced by pancreatic β-cells. [4] Diabetes is associated with chronic complications, like macrovascular diseases (e.g., cardiovascular diseases) and microvascular diseases (e.g., damages in nerve, kidney, eye, and foot). [5,6] More than 537 million adults are currently living with diabetes, and this number is expected to increase to 693 million by 2045. In 2021, 6.7 million deaths worldwide were attributed to diabetes, [7] which made it the ninth leading cause of death. [8] Globally, the prevalence of diabetes also brings a heavy burden to healthcare systems, with an estimated expenditure of over 966 billion U.S. dollars per year. [7] Diabetes is generally categorized into two groups: Type I diabetes mellitus (T1DM) and Type II diabetes mellitus (T2DM), but there are still some cases that have different etiologies, such as gestational diabetes mellitus and monogenic diabetes. [9] Since the discovery of insulin in 1921, [10] significant progress has been made in diabetes pharmacotherapy and therapeutic technology; [11][12][13] however, since a permanent cure is unavailable, the discovery and development of anti-diabetic drugs is still at the forefront of diabetes research. With the rapid advancement in high-throughput omics technologies, combined with accumulated genome-wide association study (GWAS) data, a better understanding of diabetes pathophysiology may one day lead to the discovery of novel therapeutic targets. [14,15] This review will During the past decades, unprecedented progress in technologies has revolutionized traditional research methodologies. Among these, advances in highthroughput drug screening approaches have permitted the rapid identification of potential therapeutic agents from drug libraries that contain thousands or millions of molecules. Moreover, high-throughput-based therapeutic target discovery strategies can comprehensively interrogate relationships between biomolecules (e.g., gene, RNA, and protein) and diseas...

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Targeting N‐Methyl‐lysine Histone Demethylase KDM4 in Cancer: Natural Products Inhibitors as a Driving Force for Epigenetic Drug Discovery

Cursaro,

Milioni,

Eslami

et al. 2024

ChemMedChem

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KDM4A‐F enzymes are a subfamily of histone demethylases containing the Jumonji C domain (JmjC) using Fe(II) and 2‐oxoglutarate for their catalytic function. KDM4 enzymes are overexpressed or deregulated in cancer, thus leading to alteration in chromatin structure and transcriptional defects. As KDM4 enzymes have been associated with malignancy, they may represent novel targets for developing innovative therapeutic tools to treat different solid and blood tumors, of note, KDM4A is the isozyme most frequently associated with aggressive phenotypes of these tumors. To this aim, industrial and academic medicinal chemistry efforts have identified different KDM4 inhibitors. In most cases, these are pan‐KDM4 inhibitors, with low selectivity against other Jumonji family members. The pharmacophoric features of the inhibitors frequently include a chelating group capable of coordinating the catalytic iron within the active site of the KDM4 enzyme. Nonetheless, non‐chelating compounds have also demonstrated promising inhibitory activity, suggesting potential flexibility in the drug design. Several natural products, containing monovalent or bivalent chelators, have been identified as KDM4 inhibitors, albeit with a micromolar inhibition potency. This highlights the potential for leveraging them as templates for the design and synthesis of new derivatives, exploiting nature’s chemical diversity to pursue more potent and selective KDM4 inhibitors.

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17-DMAG dually inhibits Hsp90 and histone lysine demethylases in alveolar rhabdomyosarcoma

Cited by 10 publications

References 81 publications

A protocol for high-throughput screening of histone lysine demethylase 4 inhibitors using TR-FRET assay

A protocol for high-throughput screening of histone lysine demethylase 4 inhibitors using TR-FRET assay

The Application of High‐Throughput Approaches in Identifying Novel Therapeutic Targets and Agents to Treat Diabetes

Targeting N‐Methyl‐lysine Histone Demethylase KDM4 in Cancer: Natural Products Inhibitors as a Driving Force for Epigenetic Drug Discovery

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