Highly Selective Activation of Heat Shock Protein 70 by Allosteric Regulation Provides an Insight into Efficient Neuroinflammation Inhibition

Wang, Lichao; Liao, Li-Xi; Lv, Haining; Liú, Dan; Dong, Wei; Zhu, Jian; Chen, Jinfeng; Shi, Meng-Ling; Fu, Ge; Song, Xiaomin; Jiang, Yong; Zeng, Ke‐Wu; Tu, Peng‐Fei

doi:10.1016/j.ebiom.2017.08.011

Cited by 39 publications

(44 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Human monocyte-derived macrophages transfected with Hsp70 followed by exposure to LPS expressed less TNF-α, IL-1β, IL-12 and IL-10 at mRNA level, but not IL-6, compared to macrophages transfected with Hsp70 anti-sense DNA [22]. Nevertheless, in the murine microglial cell line BV-2, the pharmacological activation of Hsp70 by handelin significantly blocked the secretion of the pro-inflammatory cytokines TNF-α, IL-1β and IL-6 [19]. In our study of canine macrophage cells, we found that the LPS-induced upregulation of TNF-α and IL-6 mRNA was dramatically diminished upon cell stress.…”

Section: Discussionmentioning

confidence: 94%

“…Inducible Hsp70 is a representative member of the family of HSPs, and has been largely examined for its functions in stressed human and murine macrophages [ 16 , 17 , 18 ]. The upregulation of Hsp70 by inducers such as celastrol was shown to inhibit the production of pro-inflammatory cytokines, such as TNF-α, IL-6 and IL-1β in BV2 cells [ 19 ], human retinal pigment epithelial cells [ 20 ], and human alveolar macrophages [ 21 ]. Similarly, the overexpression of Hsp70 attenuates LPS-induced cytokine expression in macrophages [ 22 ] and microglia cells [ 23 ].…”

Section: Introductionmentioning

confidence: 99%

“…In addition, the overexpression of Hsp72 reduced NF-κB DNA binding activity [ 23 ]. Some researchers also reported an interaction between Hsp70 and TRAF6, an essential activator of the NF-κB pathway [ 19 , 25 ], thereby disturbing NF-κB activation and translocation. Those findings suggest that Hsp70 modulates NF-κB activity.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Hsp70 and NF-kB Mediated Control of Innate Inflammatory Responses in a Canine Macrophage Cell Line

Lyu

Wawrzyniuk

Rutten

et al. 2020

IJMS

View full text Add to dashboard Cite

The pathogenesis of many inflammatory diseases is associated with the uncontrolled activation of nuclear factor kappa B (NF-κB) in macrophages. Previous studies have shown that in various cell types, heat shock protein 70 (Hsp70) plays a crucial role in controlling NF-κB activity. So far, little is known about the role of Hsp70 in canine inflammatory processes. In this study we investigated the potential anti-inflammatory effects of Hsp70 in canine macrophages as well as the mechanisms underlying these effects. To this end, a canine macrophage cell line was stressed with arsenite, a chemical stressor, which upregulated Hsp70 expression as detected by flow cytometry and qPCR. A gene-edited version of this macrophage cell line lacking inducible Hsp70 was generated using CRISPR-Cas9 technology. To determine the effects of Hsp70 on macrophage inflammatory properties, arsenite-stressed wild-type and Hsp70 knockout macrophages were exposed to lipopolysaccharide (LPS), and the expression of the inflammatory cytokines IL-6, IL-1β and tumor necrosis factor-α (TNF-α) and levels of phosphorylated NF-κB were determined by qPCR and Western Blotting, respectively. Our results show that non-toxic concentrations of arsenite induced Hsp70 expression in canine macrophages; Hsp70 upregulation significantly inhibited the LPS-induced expression of the pro-inflammatory mediators TNF-α and IL-6, as well as NF-κB activation in canine macrophages. Furthermore, the gene editing of inducible Hsp70 by CRISPR-Cas9-mediated gene editing neutralized this inhibitory effect of cell stress on NF-κB activation and pro-inflammatory cytokine expression. Collectively, our study reveals that Hsp70 may regulate inflammatory responses through NF-κB activation and cytokine expression in canine macrophages.

show abstract

Section: Discussionmentioning

confidence: 94%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Hsp70 and NF-kB Mediated Control of Innate Inflammatory Responses in a Canine Macrophage Cell Line

Lyu

Wawrzyniuk

Rutten

et al. 2020

IJMS

View full text Add to dashboard Cite

show abstract

“…OGD/R, MTT, LDH release, Hoechst 33258 staining, AO/EB staining, Western blotting, transmission electron microscope, tryptophan fluorescence quenching study, Co-IP, DARTS, mitochondrial depolarization, and CD analysis assays were performed as previously described 59-60.…”

Section: Methodsmentioning

confidence: 99%

Allosteric regulation of protein 14-3-3ζ scaffold by small-molecule editing modulates histone H3 post-translational modifications

Wan¹,

Liao²,

Liu³

et al. 2020

Theranostics

Self Cite

View full text Add to dashboard Cite

Background: Histone post-translational modifications (PTMs) are involved in various biological processes such as transcriptional activation, chromosome packaging, and DNA repair. Previous studies mainly focused on PTMs by directly targeting histone-modifying enzymes such as HDACs and HATs.Methods and Results: In this study, we discovered a previously unexplored regulation mechanism for histone PTMs by targeting transcription regulation factor 14-3-3ζ. Mechanistic studies revealed 14-3-3ζ dimerization as a key prerequisite, which could be dynamically induced via an allosteric effect. The selective inhibition of 14-3-3ζ dimer interaction with histone H3 modulated histone H3 PTMs by exposing specific modification sites including acetylation, trimethylation, and phosphorylation, and reprogrammed gene transcription profiles for autophagy-lysosome function and endoplasmic reticulum stress.Conclusion: Our findings demonstrate the feasibility of editing histone PTM patterns by targeting transcription regulation factor 14-3-3ζ, and provide a distinctive PTM editing strategy which differs from current histone modification approaches.

show abstract

“…Hsp70 has been implicated in T cell regulation of various chronic inflammatory diseases, including rheumatoid arthritis (Van Eden et al 2005), colitis (Tanaka et al 2007), neurodegenerative diseases (Van Noort 2008), and experimental autoimmune uveoretinitis (Kitamei et al 2007), which makes the molecule a potential therapeutic target in chronic inflammations. Moreover, overexpression of Hsp70 was reported to play an essential role in regulation of apoptosis (Beere et al 2000;Saleh et al 2000) and protection against inflammation (Wang et al 2017).…”

Section: Introductionmentioning

confidence: 99%

Leucinostatin acts as a co-inducer for heat shock protein 70 in cultured canine retinal pigment epithelial cells

Lyu

Ludwig

Kooten

et al. 2020

Cell Stress and Chaperones

View full text Add to dashboard Cite

Dysregulation of retinal pigment epithelium (RPE) cells is the main cause of a variety of ocular diseases. Potentially heat shock proteins, by preventing molecular and cellular damage and modulating inflammatory disease, may exert a protective role in eye disease. In particular, the inducible form of heat shock protein 70 (Hsp70) is widely upregulated in inflamed tissues, and in vivo upregulation of Hsp70 expression by HSP co-inducing compounds has been shown to be a potential therapeutic strategy for inflammatory diseases. In order to gain further understanding of the potential protective effects of Hsp70 in RPE cells, we developed a method for isolation and culture of canine RPE cells. Identity of RPE cells was confirmed by detection of its specific marker, RPE65, in qPCR, flow cytometry, and immunocytochemistry analysis. The ability of RPE cells to express Hsp70 upon experimental induction of cell stress, by arsenite, was analyzed by flow cytometry. Finally, in search of a potential Hsp70 coinducer, we investigated whether the compound leucinostatin could enhance Hsp70 expression in stressed RPE cells. Canine RPE cells were isolated and cultured successfully. Purity of cells that strongly expressed RPE65 was over 90%. Arsenite-induced stress led to a time-and dose-dependent increase in Hsp70 expression in canine RPE cells in vitro. In addition, leucinostatin, which enhanced heat shock factor-1-induced transcription from the heat shock promoter in DNAJB1-luc-O23 reporter cell line, also enhanced Hsp70 expression in arsenite-stressed RPE cells, in a dose-dependent fashion. These findings demonstrate that leucinostatin can boost Hsp70 expression in canine RPE cells, most likely by activating heat shock factor-1, suggesting that leucinostatin might be applied as a new co-inducer for Hsp70 expression.

show abstract

Highly Selective Activation of Heat Shock Protein 70 by Allosteric Regulation Provides an Insight into Efficient Neuroinflammation Inhibition

Cited by 39 publications

References 45 publications

Hsp70 and NF-kB Mediated Control of Innate Inflammatory Responses in a Canine Macrophage Cell Line

Hsp70 and NF-kB Mediated Control of Innate Inflammatory Responses in a Canine Macrophage Cell Line

Allosteric regulation of protein 14-3-3ζ scaffold by small-molecule editing modulates histone H3 post-translational modifications

Leucinostatin acts as a co-inducer for heat shock protein 70 in cultured canine retinal pigment epithelial cells

Contact Info

Product

Resources

About