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Updated: May 22, 2025
Studies of Chaperone-Cochaperone Interactions using Homogenous Bead-Based Assay
Published on: July 21, 2021
Identification of novel small molecule chaperone activators for neurodegenerative disease treatment
Anita K Ho1, Fiona Jeganathan2, Magda Bictash2
1York Biomedical Research Institute, Department of Biology, University of York, Wentworth Way, York YO10 5DD, UK.
Abstract:
A pathological hallmark of neurodegenerative disease is the accumulation of aberrant protein aggregates which contribute to the cytotoxicity and are therefore a target for therapy development. One key mechanism to manage cellular protein homeostasis is heat shock proteins (HSPs), protein chaperones which are known to target aberrant protein accumulation. Activation of HSPs target aberrant TDP-43, tau and amyloid to rescue neurodegenerative disease. As an attempt to target HSP activation for neurodegeneration therapy, we here develop a drug screening assay to identify compounds that will activate the master regulator of HSPs, the transcription factor heat shock factor 1 (HSF1). As HSF1 is bound by HSP90 which prevents its activation, we developed a NanoBRET assay, which allows us to monitor and quantify the HSF1-HSP90 interaction in living cells to screen for compounds disrupting this interaction and thereby releasing HSF1 for activation. After the optimisation and validation of the assay, a two thousand compound library was screened which produced 10 hits including two known HSP90 inhibitors. Follow-up functional study showed that one of the hits oxyphenbutazone (OPB) significantly reduces the accumulation of insoluble TDP-43 in a cell model, eliciting no signs of stress or toxicity. Overall, this study demonstrates a viable strategy for new drug discovery in targeting aberrant proteins and identifies potential candidates for translation into neurodegenerative disease treatment.
Insights
Researchers developed a drug screening assay to activate heat shock factor 1 (HSF1), a key regulator of cellular protein homeostasis, to combat neurodegenerative diseases. The study identified oxyphenbutazone as a promising compound that reduces toxic protein aggregates without causing cellular stress.
Area of Science:
- Neuroscience
- Molecular Biology
- Drug Discovery
Background:
- Aberrant protein aggregates are pathological hallmarks of neurodegenerative diseases, contributing to cytotoxicity.
- Heat shock proteins (HSPs) are crucial for maintaining cellular protein homeostasis and can target toxic protein accumulations.
- Activating HSPs offers a therapeutic strategy to mitigate neurodegenerative conditions by clearing aberrant proteins like TDP-43, tau, and amyloid.
Purpose of the Study:
- To develop a drug screening assay for identifying compounds that activate heat shock factor 1 (HSF1), the master regulator of HSPs.
- To screen for compounds that disrupt the HSF1-HSP90 interaction, thereby releasing HSF1 for activation.
- To identify novel therapeutic candidates for neurodegenerative diseases by targeting aberrant protein aggregation.
Main Methods:
- Development and validation of a NanoBRET assay to quantify the HSF1-HSP90 interaction in living cells.
- Screening of a two thousand compound library using the developed assay.
- Functional studies of hit compounds in a cellular model of neurodegeneration.
Main Results:
- The NanoBRET assay successfully monitored and quantified the HSF1-HSP90 interaction.
- Screening identified 10 hit compounds, including two known HSP90 inhibitors.
- Oxyphenbutazone (OPB), a novel hit, significantly reduced insoluble TDP-43 accumulation in a cell model without inducing toxicity.
Conclusions:
- The study presents a viable drug discovery strategy targeting aberrant protein aggregation in neurodegenerative diseases.
- The developed assay is effective for identifying compounds that activate HSF1.
- Oxyphenbutazone shows therapeutic potential for treating neurodegenerative conditions characterized by TDP-43 aggregation.
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