Abstract
Neurons are susceptible to different cellular stresses and this vulnerability has been implicated in the pathogenesis of Huntington’s disease (HD). Accumulating evidence suggest that acute or chronic stress, depending on its duration and severity, can cause irreversible cellular damages to HD neurons, which contributes to neurodegeneration. In contrast, how normal and HD neurons respond during the resolution of a cellular stress remain less explored. In this study, we challenged normal and HD cells with a low-level acute ER stress and examined the molecular and cellular responses after stress removal. Using both striatal cell lines and primary neurons, we first showed the temporal activation of p-eIF2α-ATF4-GADD34 pathway in response to the acute ER stress and during recovery between normal and HD cells. HD cells were more vulnerable to cell death during stress recovery and were associated with increased number of apoptotic/necrotic cells and decreased cell proliferation. This is also supported by the Gene Ontology analysis from the RNA-seq data which indicated that “apoptosis-related Biological Processes” were more enriched in HD cells during stress recovery. We further showed that HD cells were defective in restoring global protein synthesis during stress recovery and promoting protein synthesis by an integrated stress response inhibitor, ISRIB, could attenuate cell death in HD cells. Together, these data suggest that normal and HD cells undergo distinct mechanisms of transcriptional reprogramming, leading to different cell fate decisions during the stress recovery.
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The authors declare that the data supporting the findings of this study are available within the article and its supplementary information files.
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Funding
This work was supported by the National Institutes of Health (NS111202 to J.W.) and Florida Department of Health (9AZ06 to J.W.). J.W. was partially supported by the National Institutes of Health (EB025819).
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HX performed immunostaining, Western blot and proliferation assay. JB and HX prepared the RNA samples for RNA-seq. EC and JW performed the RNA-seq data analysis. JW and MLL planned the experiment, analyzed the data and drafted the first manuscript. All authors read and approved the final manuscript.
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Mice were handled in accordance with the animal protocol approved by the Institutional Animal Care and Use Committees (IACUC) at Florida Atlantic University. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.
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10571_2021_1137_MOESM1_ESM.tif
Supplementary file1 Supplementary Fig. S1. A. Quantitative densitometry analysis of total eIF2α levels normalized to actin during stress (N=3-7 samples per group). B. 100 nM Tg treatment for 30min did not lead to ATF6 cleavage as analyzed by Western blot. C. Quantitative RT-PCR analysis showed that 100 nM Tg treatment did not induce XBP1 mRNA splicing. N=5-6 samples per group. One-way ANOVA with Tukey post hoc analysis. D. Quantitative densitometry analysis of total eIF2α levels normalized to actin during stress recovery (N=3-8 samples per group). (TIF 11123 kb)
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Supplementary file2 Supplementary Fig. S2. Nuclear localization of ATF4 was increased during stress recovery in both STHdhQ7 and Q111 cells. Scale bar: 10 μm. (TIF 13083 kb)
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Supplementary file3 Supplementary Fig. S3. ISRIB attenuated Tg-induced SG formation in STHdhQ111 cells. Representative confocal images showing SG formation under Tg treatment in the absence (A) or presence (B) of ISRIB. Nuclei were counterstained with DAPI. Scale bar 20 μm. (TIF 36179 kb)
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Xu, H., Bensalel, J., Capobianco, E. et al. Impaired Restoration of Global Protein Synthesis Contributes to Increased Vulnerability to Acute ER Stress Recovery in Huntington’s Disease. Cell Mol Neurobiol 42, 2757–2771 (2022). https://doi.org/10.1007/s10571-021-01137-9
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DOI: https://doi.org/10.1007/s10571-021-01137-9