Induction Of Heat Shock Proteins Research Articles

Enhancing resistance and cell survival in Acipenser ruthenus liver, gill, and kidney cells: The potential of heat shock protein inducers against PAH-benzo[a]pyrene stress.

The Caspian Sea has faced many environmental challenges, such as oil pollution. Heat shock proteins (HSPs) play a critical role in stress conditions and physiological changes caused by disease or injury. By evaluating the effects of various HSP inducers (HSPi), including Pro-Tex® (NOP: 800 mM), amygdalin (AMG: 80 mM), and a novel synthetic compound derived from pirano piranazole (SZ: 80 µm) on isolated cells from Sterlet Sturgeon (Acipenser ruthenus) treated with 75% IC50 PAH-benzo[a]pyrene (BaP; B75). This study examines whether there is a correlation between exposure to the BaP pollutant and HSPs in fish. In vitro, after culturing cells from the liver, kidney, and gills, they were treated with HSPi compounds in the presence and absence of BaP. Western blotting was used to assess HSP27, HSP70, and HSP90 expression patterns. A variety of enzyme activities were measured before (without treatment) and after treatment with HSPis and HSPi + B75, including cytochrome P450 (CYP450) activity, specific enzyme activity for acetylcholinesterase (AChE), antioxidant capacity, liver indicator enzymes, cortisol levels, and immunity parameters. When compared to the control group, cells treated with B75 showed the lowest AChE enzyme activity (p < 0.0001). CYP450 activity was highest in group B75, while HSPi caused the opposite effect (p < 0.0001). HSPi + B75 increased HSP levels and antioxidant parameters while decreasing cortisol and liver indicator enzymes (p < 0.0001). HSPi may be a powerful and reliable method for enhancing the resistance of A. ruthenus to BaP stresses before exposure. Treating cells with HSP-inducing compounds, such as NOP, AMG, and SZ, can assist them in managing stress and increase HSP (27, 70, and 90) protein expression. Furthermore, the study findings suggest that HSPis can also mitigate the adverse effects of stress, ultimately increasing cell survival and resistance.

Potential Effects of Ischemic Postconditioning and Changes in Heat Shock Protein 72 in Patients with Acute Myocardial Infarction without Prodromal Angina.

The effectiveness of ischemic postconditioning (iPoC) in patients with ST-elevation myocardial infarction (STEMI) without ischemic preconditioning has not been determined. Therefore, we investigated the impact of iPoC and its potential mechanism related to heat shock protein 72 (HSP72) induction on myocardial salvage in patients with STEMI without prodromal angina (PA).We retrospectively analyzed data from 102 patients with STEMI with successful reperfusion among 323 consecutive patients with acute coronary syndrome. Among these, 55 patients with iPoC (iPoC (+) ) underwent 4 cycles of 60-second inflation and 30-second deflation of the angioplasty balloon. Both the iPoC (+) and iPoC (-) groups were divided into 2 further subgroups: patients with PA (PA (+) ) and those without (PA (-) ). We analyzed HSP72 levels in neutrophils, which were measured until 48 hours after reperfusion. I-123 β-methyl-p-iodophenyl-pentadecanoic acid (BMIPP) scintigraphy was performed within a week of reperfusion therapy. In 64% of patients, thallium-201 (TL) scintigraphy was performed 6-8 months after STEMI onset.Using BMIPP and TL, in the PA (-) subgroups, the iPoC (+) group had a significantly greater myocardial salvage ratio than the iPoC (-) group. iPoC was identified as an independent predictor of the myocardial salvage ratio. The HSP72 increase ratio was significantly elevated in the iPoC (+) PA (-) group. Importantly, the myocardial salvage effect in patients without PA was significantly correlated with the HSP72 increase ratio, which was greater in patients with iPoC.These results suggest the potential impact of iPoC via HSP72 induction on myocardial salvage; however, the effects may be limited to patients with STEMI without PA.

Endogenous fluorescent reporters for heat shock proteins are not detectable after stress induction.

Mitochondria and the endoplasmic reticulum (ER) utilise unique unfolded protein response (UPR) mechanisms to maintain cellular proteostasis. Heat shock proteins (HSPs) are UPR chaperones induced by specific stressors to promote protein folding. Previous research has successfully employed transgenic reporters in Caenorhabditis elegans to report HSP induction. However, transgenic reporters are overexpressed and only show promoter regulation and not post-transcriptional regulation. To examine endogenous HSP regulation, we attempted to generate and validate endogenous reporters for mitochondrial ( HSP-60 ) and ER ( HSP-4 ) chaperones. Using CRISPR/Cas9 technology, F2A-GFP-H2B coding DNA was inserted downstream of each HSP gene and stress induction assays conducted to validate these tools. Endogenous reporters were successfully generated for hsp-4 and hsp-60 . However, GFP induction could not be detected with these endogenous reporters upon stress induction, likely due to low level expression.

Using heat shock protein (HSP) inducers as an approach to increase the viability of sterlet (Pisces; Acipenseridae; Acipenser ruthenus) cells against environmental diazinon toxicity

Diazinon (DZN) is an organophosphate pesticide frequently used in agriculture and released into aquatic environments. In this study, sterlet sturgeon cells were exposed to DZN to investigate possible defense mechanisms via HSP induction (HSPi). Liver, kidney, and gill cells of Acipenser ruthenus were isolated and cultured and then treated with HSPi (Pro-Tex®, amygdalin, and a novel pirano-piranazole-based synthesized compound: SZ) in the presence and absence of DZN. MTT assays were used to evaluate the effects of different HSPis and their combinations with DZN. Western blotting analysis was conducted to evaluate HSP27, HSP70, and HSP90 expression patterns in each group. The highest rates of caspase-3 and caspase-8 activities were found in the DZN group, whereas HSPi treatment resulted in the lowest rates. The combination of HSPi+DZN resulted in increased HSP levels and antioxidant parameters but decreased cortisol, immune parameters, and metabolic enzymes. Many of the studied parameters (caspases, acetylcholinesterase, antioxidant, immune, and metabolic parameters) showed significant correlations with HSP expression, indicating that HSPs may be associated with markers of sterlet cell health. The results of this study demonstrate that using HSP inducers may be a powerful and reliable way to increase A. ruthenus resistance prior to exposure to DZN.

Transcriptomic analyses of juvenile Striped Bass (Morone saxatilis) exposed to chronic and acute temperature change.

Striped Bass are economically important, migratory fishes, which occur across a wide range of latitudes. Given their wide-ranging nature, Striped Bass can cope with a broad range of environmental temperatures, yet the mechanisms underlying this ability have not been thoroughly described. Heat shock proteins (HSPs) are inducible molecular chaperones, which help mitigate protein damage resulting from increased temperatures. The importance of HSPs has been demonstrated in a number of fish species, but their role in Striped Bass is poorly understood. This study characterizes changes in gene expression in juvenile Striped Bass, following acute and chronic temperature change. Fish were acclimated to one of three temperatures (15, 25 or 30°C) and sampled at one of two treatments (control or after CTmax), following which we assessed differential gene expression and gene ontology in muscle. It is clear from our differential expression analyses that acclimation to warm temperatures elicits more robust changes to gene expression, compared to acute temperature increases. Our differential expression analyses also revealed induction of many different heat shock proteins, including hsp70, hsp90, hsp40 and other small HSPs, after both acute and chronic temperature increase in white muscle. Furthermore, the most consistent gene ontology pattern that emerged following both acclimation and CTmax was upregulation of transcripts involved in "protein folding", which also include heat shock proteins. Gene ontology analyses also suggest changes to other processes after acclimation, including decreased growth pathways and changes to DNA methylation. Overall, these data suggest that HSPs likely play a major role in the Striped Bass's ability to tolerate warm waters.

Heat stress induced in vitro affects cell viability and gene expression of dermal fibroblasts from bovine and buffalo

Aim of study: To evaluate the response of dermal fibroblasts to heat stress and different time exposures on the cell survival and gene expression. Area of study: Belém city, Pará state. Brazil. Material and methods: Fibroblasts were isolated from ear skin of bovine (n= 4) and buffalo (n= 4), cultured in vitro until the 3rd passage and submitted to heat stress at 42°C for 3, 6 and 12 h, except for the negative control (38.5°C for 24 h). Cell survival was measured using Trypan blue, and RNA isolation was performed using Trizol method following qRT-PCR to quantify the relative expression of the inducible heat shock protein HSPA1A, the pro-apoptotic BAX and pro-inflammatory IFN-γ genes. Main results: Heat stress induced in vitro affected the cell viability and gene expression in a time-dependent manner. Gene expression was relatively lower in buffalo (p&lt;0.05) than in bovine. Until 3 h of heat stress, HSPA1A showed a slight increase in both bovine and buffaloes, and BAX was 5.82-fold greater in bovine (p&lt;0.05). After 6 h, HSPA1A was 75.81-fold (p&lt;0.0001) and INF-γ was 20.15-fold greater (p&lt;0.05) in bovine than buffalo. Only after 6 h the cell viability started to decrease significantly (p&lt;0.05) in both species. Research highlights: Dermal fibroblasts of buffaloes and bovine were sensitive to heat stress induced in vitro, which was most detrimental to cell survival after 6 h. The expression of HSPA1A, BAX and INF-γ genes in response to heat stress indicate a slight sensibility of the dermal fibroblasts of bovine compared to their buffalo counterpart.

THU083 Disulfiram Mediated Anti-tumor Effect In Pituitary Neuroendocrine Tumors By Inducing Cuproptosis

Abstract Disclosure: N. Huang: None. Y. Zhang: None. Y. Liu: None. Y. Feng: None. L. Xue: None. Z. Wu: None. Pituitary neuroendocrine tumors (PitNETs) are common intracranial tumors. Dopamine agonists and somatostatin analogues are the first-line drugs for PRL and GH-secreting PitNETs clinically. However, there is no effective drug for other subtypes and drug-resistant PitNETs. Thus, it’s impending to discover new therapeutic targets and medicine. Cuproptosis is a copper-dependent form of cell death driven by lipoylated protein aggregation, loss of Fe-S cluster-containing proteins, and induction of HSP70. However, the role of cuproptosis in PitNETs is not clear. In this study, a total of 17 primary samples of PitNETs were collected to perform high-throughput drug screening, an anti-abuse drug disulfiram (DSF) was identified that can induce cell death effectively. We found that disulfiram also has a strong inhibitory effect on pituitary tumor cell lines MMQ, Att20 and GH3 (-87.1%, -62.5%, -92.7%, p&amp;lt;0.001), but the inhibitory effect was eliminated by copper chelator TTM/BCS. This indicated that DSF played the anti-tumor effect requiring the participation of copper ions. To explore the underlying mechanism that DSF induces the cell death of PitNETs, we detected the hallmark of cuproptosis and found that DSF leads to a decrease in the acylation level of proteins related to the tricarboxylic acid cycle, limiting the function of the protein. On the one hand, DSF leads to the downregulation of iron-sulfur cluster-associated proteins, and the oligomerization of dihydrolipoamide S-acetyltransferase (DLAT), which is eventually accompanied by an increase in the protein level of heat shock protein 70 (HSP70) at protein and mRNA level, causing proteotoxic stress leading to cell death. Finally, DSF treatment inhibited the tumor growth in MMQ cell transplanted mouse model and destructed the morphology of the pituitary tumor 3D sphere similarly demonstrating the inhibition effect. In conclusion, we find that the anti-abuse drug disulfiram can induce coproptosis in pituitary tumor cells in a copper-dependent manner and is a potential drug for the treatment of PitNETs. Presentation: Thursday, June 15, 2023

Involvement of the heat shock response (HSR) regulatory pathway in cadmium-elicited cerebral damage.

The heat shock response (HSR) is a cellular protective mechanism that is characterized by the induction of heat shock transcription factors (HSFs) and heat shock proteins (HSPs) in response to diverse cellular and environmental stressors, including cadmium (Cd). However, little is known about the relationship between the damaging effects of Cd and the HSR pathway in the chicken cerebrum following Cd exposure. To explore whether Cd exposure elicits cerebral damage and triggers the HSR pathway, chicks were exposed to Cd in the daily diet at different concentrations (35, 70, or 140mg/kg feed) for 90days, while a control group was fed the standard diet without Cd. Histopathological examination of cerebral tissue from Cd-exposed chickens showed neuronal damage, as evidenced by swelling and degeneration of neurons, loss of neurons, and capillary damage. Cd exposure significantly increased mRNA expression of HSF1, HSF2, and HSF3, and mRNA and protein expression of three major stress-inducible HSPs (HSP60, HSP70, and HSP90). Moreover, Cd exposure differentially modulated mRNA expression of small HSP (sHSPs), most notably reducing expression of HSP27 (HSPB1). Furthermore, Cd exposure increased TUNEL-positive neuronal apoptotic cells and up-regulated protein expression of caspase-1, caspase-8, caspase-3, and p53, leading to apoptosis. Taken together, these data demonstrate that activation of the HSR and apoptotic pathways by Cd exposure is involved in Cd-elicited cerebral damage in the chicken. Synopsis for the graphical abstract Cadmium (Cd)-induced neuronal damage triggers the heat shock response (HSR) by activating heat shock transcription factors (HSFs) and subsequent induction of major heat shock proteins (notably, HSP60, HSP70, and HSP90). Moreover, Cd exposure activates caspase-1, caspase-8, caspase-3, and p53 protein, thereby resulting in neuronal apoptosis in the chicken brain.

Lymphocyte Expression of Intracellular Cytokines and Heat Shock Proteins in Peripheral Blood of Patients with Atopic Dermatitis.

The present study evaluates expression by activated CD4+ T helper1 (Th1) and T helper 2 (Th2) T lymphocytes of pro-inflammatory cytokines and cytoprotective heat shock proteins (HSPs) in peripheral blood of atopic dermatitis (AD) patients. This research represents preliminary work by the authors to identify correlates between critical immune parameters with the potential to serve as guidelines for the development of pharmacological strategies for altering these factors to promote the restoration of healthy immune profiles in persons afflicted with major atopic diseases. The major experimental strategy used in this research assessed immune activation by peripheral blood mononuclear cells (PBMC) from 21 AD patients and 12 age- and gender-matched healthy control subjects cultured with phorbol myristate acetate (PMA) and ionomycin (PMA/I), which are mutagenic immune activators, to induce expression of pro-inflammatory biomarkers in CD4+ T cells differentiated to express Th1 or Th2 cytokines and heme oxygenase-1 (HO-1) intracellularly (i). Evaluations were performed using an FC500 Beckman-Coulter flow cytometer. Elevated CD4+ T cell expression of cytokines, interleukin-4 (iIL-4), interleukin- 5 (iIL-5), interleukin-10 (iIL-10), interferon-gamma (iIFN-g), tumor necrosis factor-alpha (iTNF-α), were observed. Additionally, the heat shock proteins (HSP) iHO-1 and iHSP-70 were evaluated in cells from the blood of AD patients versus the control subjects. The present study demonstrated an elevated expression of both Th1 and Th2-associated cytokines in CD4+ T cells of AD patients, with a significant direct correlation between Th1 and Th2 cell populations, thus yielding insight into the immune features of the AD-associated systemic inflammatory profile. Finally, the observed increased iHO-1 and iHSP-70 expressions likely represent adaptive physiologic countermeasures to AD-associated inflammatory tissue damage, suggesting that HSP inducers are promising candidates for the management of atopic disorders.

Greater physiological resistance to heat may favour an invasive freshwater turtle, enabling it to outcompete native species in a changing climate

Abstract Climate warming and biological invasion can interact to affect ecosystems, and the results of such interactions depend on whether invaders show physiological advantages in their responses to warming environments as compared with native species after the establishment and local adaptation. However, it has rarely been evaluated. The present study compared the heat tolerance of a globally invasive turtle, the red‐eared slider Trachemys scripta elegans, and two co‐occurring native turtle species in China, the Chinese soft‐shelled turtle Pelodiscus sinensis and Chinese pond turtle Mauremys reevesii, after acclimation in the same thermal environment. The expression patterns of heat shock factor 1 (HSF1), multiple heat shock proteins (HSPs) and telomere reverse transcriptase (TERT), and changes in relative telomere length (RTL) were then evaluated in liver and muscle of turtles under thermal stress. The results showed that T. scripta elegans had greater physiological resistance to heat than the co‐occurring native turtle species, and that this higher tolerance was accompanied by a higher temperature induction of the peak expression of inducible HSPs in liver and a higher onset temperature for the main inducible HSPs in muscle. During thermal stress, T. scripta elegans showed a better ability to maintain the expression levels of TERT under extremely hot conditions, whereas high temperatures inhibited TERT in muscles of native turtle species. Meanwhile, thermal stress did not cause telomere erosion in any of the three turtle species and some treatments can lead to slightly longer telomeres in T. scripta elegans or M. reevesii. Overall, the invasive turtle species showed higher heat tolerance and physiological advantages in enduring hot weather in comparison to the co‐occurring native turtle species. This may favour these invaders in situations of climate warming. Our study highlights the role of physiological adaptations displayed by invaders in achieving advantages over native species under climate change.

GGA (geranylgeranylacetone) ameliorates bleomycin-induced lung inflammation and pulmonary fibrosis by inhibiting apoptosis and oxidative stress.

Fibrosis is a response to ongoing cellular injury, disruption, and tissue remodeling, the pathogenesis of which is unknown, and is characterized by extracellular matrix deposition. The antifibrotic effect of Geranylgeranylacetone (GGA), as an inducer of Heat shock protein 70 (HSP70), in liver, kidney and pulmonary fibrosis has been supported by multiple preclinical evidence. However, despite advances in our understanding, the precise roles of HSP70 in fibrosis require further investigation. The purpose of this study was to investigate whether GGA could participate in the progression of pulmonary fibrosis in mice through apoptosis, oxidative stress and inflammation. B-cell lymphoma-2(Bcl-2) and Bcl2-Associated X (Bax) are two proteins related to apoptosis. Anti-apoptotic factor Bcl-2 and pro-apoptotic factor Bax are often involved in the apoptotic process in the form of dimer. Immunofluorescence and Western blot results showed that bleomycin (BLM) and transforming growth factor-β (TGF-β) inhibited Bcl-2 expression and promoted Bax expression in vitro and in vivo, respectively. In contrast, GGA treatment reverses this change. Reactive oxygen species (ROS), Malondialdehyde (MDA) and superoxide dismutase (SOD) are markers of oxidative stress, which often reflect oxidative injury of cells. The detection of ROS, MDA and SOD expression showed that TGF-β and BLM treatment could significantly promote oxidative stress, while GGA treatment could alleviate oxidative stress damage. In addition, BLM significantly elevated Tumor necrosis factor-α(TNF-α), Interleukin1β (IL-1β) and Interleukin 6 (IL-6), while scutellarin reversed the above alterations except for that of GGA. Taken together, GGA suppressed apoptotic, oxidative stress and inflammation in BLM-induced pulmonary fibrosis.

The Probiotic Phaeobacter inhibens Provokes Hypertrophic Growth via Activation of the IGF-1/Akt Pathway during the Process of Metamorphosis of Greater Amberjack (Seriola dumerili, Risso 1810)

Metamorphosis entails hormonally regulated morphological and physiological changes requiring high energy levels. Probiotics as feed supplements generate ameliorative effects on host nutrient digestion and absorption. Thereby, the aim of the present research was to investigate the impact of the probiotic Phaeobacter inhibens as a water additive on cellular signaling pathways in the metamorphosis of greater amberjack (Seriola dumerili). Activation of insulin-like growth factor type 1 receptor (IGF-1R), protein kinase B (Akt), mitogen-activated protein kinases (MAPKs) and AMP-activated protein kinase (AMPK), induction of heat shock proteins (Hsps), and programmed cell death were assessed through SDS-Page/immunoblot analysis, while energy metabolism was determined through enzymatic activities. According to the results, greater amberjack reared in P. inhibens-enriched water entered the metamorphic phase with greater body length, while protein synthesis was triggered to facilitate the hypertrophic growth as indicated by IGF-1/Akt activation and AMPK inhibition. Contrarily, MAPKs levels were reduced, whereas variations in Hsps response were evident in the probiotic treatment. Apoptosis and autophagy were mobilized potentially for the structural remodeling processes. Furthermore, the elevated enzymatic activities of intermediary metabolism highlighted the excess energy demands of metamorphosis. Collectively, the present findings demonstrate that P. inhibens may reinforce nutrient utilization, thus leading greater amberjack to an advanced growth and developmental state.

Geranylgeranylacetone, an inducer of heat shock protein 70, attenuates pulmonary fibrosis via inhibiting NF-κB/NOX4/ROS signalling pathway in vitro and in vivo

Idiopathic pulmonary fibrosis (IPF) is a devastating and progressive pulmonary disease which is characterized by epithelial cell damage and extracellular collagen deposition. To date, the therapeutic options for IPF are still very limited, so the relevant mechanisms need to be explored. Heat shock protein 70 (HSP70), which has protective versus antitumor effects on cells under stress, is a member of the heat shock protein family. In the current study, qRT–PCR, western blotting, immunofluorescence staining, and migration assays were used to explore the Epithelial-mesenchymal transition (EMT) process in BEAS-2B cells. Moreover, the role of GGA in the process of pulmonary fibrosis was detected by HE, Masson staining, pulmonary function test and immunohistochemistry in C57BL/6 mice. Our results indicated that GGA, as an inducer of HSP70, enhanced the transformation of BEAS-2B cells from epithelial to mesenchymal cells through the NF-κB/NOX4/ROS (reactive oxygen species) signalling pathway and could significantly reduce apoptosis of BEAS-2B cells induced by TGF-β1(Transforming growth factor β1) in vitro. In vivo studies demonstrated that HSP70-inducing drugs, such as GGA, attenuated pulmonary fibrosis progression induced by bleomycin (BLM). Collectively, these results suggested that overexpression of HSP70 attenuated pulmonary fibrosis induced by BLM in C57BL/6 mice and EMT process induced by TGF-β1 through NF-κB/NOX4/ROS pathway in vitro. Thus, HSP70 might be a potential therapeutic strategy for human lung fibrosis.

Expression of HSP70 in Human Skin After Cryolipolysis Treatment.

Cryolipolysis nonsurgically targets and reduces subcutaneous fat through controlled cooling of skin and underlying fatty tissue. Although skin changes after cryolipolysis treatment have been observed clinically, the mechanisms by which these occur are not well understood. The aim of this study was to investigate the expression of heat shock protein 70 (HSP70) in the epidermal and dermal layers of human skin following cryolipolysis treatment. Subjects (N = 11; average age, 41.8 years; average BMI, 29.59 kg/m2) were recruited to receive cryolipolysis treatment with a vacuum cooling cup applicator (-11°C/35 minutes) prior to abdominoplasty surgery. Treated and untreated abdominal tissue samples were harvested immediately after surgery (average follow-up, 15 days; range, 3 days to 5 weeks). Immunohistochemistry for HSP70 was performed on all samples. Slides were digitized and quantified in epidermal and dermal layers. There was higher epidermal and dermal HSP70 expression in cryolipolysis-treated pre-abdominoplasty samples vs untreated samples. There was a 1.32-fold increase of HSP70 expression in the epidermis (P < .05) and a 1.92-fold increase in the dermis (P < .04) compared with untreated samples. We found significant induction of HSP70 after cryolipolysis treatment in epidermal and dermal layers. HSP70 has potential therapeutic benefits and is recognized to have a role in skin protection and adaption after thermal stress. Although cryolipolysis is popular for subcutaneous fat reduction, cryolipolytic HSP induction in the skin may prove valuable for additional therapeutic applications, including skin wound healing, remodeling, rejuvenation, and photoprotection.

Heat shock factor 1 is a promising therapeutic target against adult T-cell leukemia

Patients with adult T-cell leukemia (ATL), which is caused by human T-cell leukemia virus type 1 (HTLV-1), show poor prognosis because of drug resistance. Heat shock protein (HSP) 90 is reportedly essential for ATL cell survival as it regulates important signaling pathways, thereby making HSP90 inhibitors new therapeutic candidates for ATL. However, HSP90 inhibition increases the expression of other HSPs, suggesting that HSPs may contribute to drug resistance. The heat shock factor 1 (HSF1) transcription factor is the primary regulator of the expression of HSPs. Furthermore, targeting HSF1 disrupts the HSP90 chaperone function. Herein, we demonstrated that HSF1 is overexpressed in HTLV-1-infected T cells. HSF1 knockdown inhibited the proliferation of HTLV-1-infected T cells. HSF1 inhibitor KRIBB11 reduced the expression and phosphorylation of HSF1, downregulated HSP70 and HSP27 expression, and suppressed Akt, nuclear factor-κB, and AP-1 signals. KRIBB11 treatment induced DNA damage, upregulated p53 and p21, and reduced the expression of cyclin D2/E, CDK2/4, c-Myc, MDM2, and β-catenin, thereby preventing retinoblastoma protein phosphorylation and inhibiting G1-S cell cycle progression. KRIBB11 also induced caspase-mediated apoptosis concomitant with the suppression of Bcl-xL, Mcl-1, XIAP, c-IAP1/2, and survivin expression. KRIBB11 inhibited HSP70 and HSP90 upregulation through treatment with AUY922, an HSP90 inhibitor, and enhanced the cytotoxic effect of AUY922, suggesting a salvage role of HSF1-dependent HSP induction in response to drug treatment. Finally, treatment of mice with KRIBB11 reduced ATL tumor growth. Therefore, this study provides a strong rationale to target HSF1 and validates the anti-ATL activity of KRIBB11.

Chronic heat stress induces the expression of HSP genes in the retina of chickens (Gallus gallus).

Introduction: Chronic heat stress during summer is a major challenge imposed by global warming. Chickens are more sensitive to heat stress than mammals because they lack sweat glands. Thus, chickens are more susceptible to heat stress during summer than other seasons. Induction of heat shock protein (HSP) genes is one of the primary defense mechanisms against heat stress. Tissue-specific responses exhibited by different classes of HSPs upon exposure to heat stress have been reported previously in different tissues including the heart, kidney, intestine, blood, and muscle, but not in the retina. Therefore, this study aimed to investigate the expression levels of HSP27, HSP40, HSP60, HSP70, and HSP90 in the retina under chronic heat stress. Methods: This study was conducted during the summers of 2020 and 2021 in Kuwait. Chickens (Gallus gallus) were divided into control and heat-treated groups and sacrificed at different developmental stages. Retinas were extracted and analyzed by using Real Time quantitative Polymerase Chain Reaction (RT-qPCR). Results: Our results from the summer of 2021 were similar to that from the summer of 2020, regardless of whether GAPDH or RPL5 was used as a gene normalizer. All five HSP genes were upregulated in the retina of 21-day-old heat-treated chickens and stayed upregulated until 35days of age, with the exception of HSP40, which was downregulated. The addition of two more developmental stages in the summer of 2021 showed that at 14days, all HSP genes were upregulated in the retina of heat-treated chickens. In contrast, at 28days, HSP27 and HSP40 were downregulated, whereas HSP60, HSP70, and HSP90 were upregulated. Furthermore, our results showed that under chronic heat stress, the highest upregulation of HSP genes was seen at the earliest developmental stages. Discussion: To the best of our knowledge, this is the first study to report the expression levels of HSP27, HSP40, HSP60, HSP70, and HSP90 in the retina under chronic heat stress. Some of our results match the previously reported expression levels of some HSPs in other tissues under heat stress. These results suggest that HSP gene expression can be used as a biomarker for chronic heat stress in the retina.

SATB1 Chromatin Loops Regulate Megakaryocyte/Erythroid Progenitor Expansion by Facilitating HSP70 and GATA1 Induction.

Diamond Blackfan anemia (DBA) is an inherited bone marrow failure syndrome associated with severe anemia, congenital malformations, and an increased risk of developing cancer. The chromatin-binding special AT-rich sequence-binding protein-1 (SATB1) is downregulated in megakaryocyte/erythroid progenitors (MEPs) in patients and cell models of DBA, leading to a reduction in MEP expansion. Here we demonstrate that SATB1 expression is required for the upregulation of the critical erythroid factors heat shock protein 70 (HSP70) and GATA1 which accompanies MEP differentiation. SATB1 binding to specific sites surrounding the HSP70 genes promotes chromatin loops that are required for the induction of HSP70, which, in turn, promotes GATA1 induction. This demonstrates that SATB1, although gradually downregulated during myelopoiesis, maintains a biological function in early myeloid progenitors.

Comparison of BiP and HSP70i as markers of unfolded protein response (UPR) in segmental and nonsegmental vitiligo

Background: Unfolded protein response (UPR) is a misfolded protein that occurs because oxidative stress disrupts cellular redox potential that extends to the endoplasmic reticulum (ER). Binding immunoglobulin protein (BiP) and inducible heat shock protein (HSP70i) as ER chaperons play critical roles in melanocyte apoptosis. Our study aims to compare BiP and HSP70i as markers of UPR in patients with segmental vitiligo (SV) and non-segmental vitiligo (NSV). Methods: The subjects were composed of 64 patients diagnosed with vitiligo, of whom 33 had NSV and 31 had SV. Skin biopsy and immunofluorescence were performed. We used BiP and HSP70i as markers of UPR. Descriptive statistics were used to analyze the data. Results: UPR-BiP expression and UPR-HSP70i in the SV group was 2.66 ± 3.07 and 3.85 ± 4.92, respectively, with a p-value of 0.001 (&lt;α = 0.05). In the NSV group UPR-BiP expression and UPR-HSP70i was 12.55 ± 11.85 and 14.79 ± 14.72 respectively, with a p-value of 0.001 (&lt;α = 0.05). UPR expression using the HSP70i marker in both NSV and SV groups was higher than it was using the BiP marker. The relationship between UPR-HSP70i expression and UPR-BiP expression in the SV and NSV group was significant (p &lt;0.05) and positive. Conclusions: HSP70i is superior to BiP as a marker for expressing UPR.

HSP70 Family in Cancer: Signaling Mechanisms and Therapeutic Advances.

The 70 kDa heat shock proteins (HSP70s) are a group of highly conserved and inducible heat shock proteins. One of the main functions of HSP70s is to act as molecular chaperones that are involved in a large variety of cellular protein folding and remodeling processes. HSP70s are found to be over-expressed and may serve as prognostic markers in many types of cancers. HSP70s are also involved in most of the molecular processes of cancer hallmarks as well as the growth and survival of cancer cells. In fact, many effects of HSP70s on cancer cells are not only related to their chaperone activities but rather to their roles in regulating cancer cell signaling. Therefore, a number of drugs directly or indirectly targeting HSP70s, and their co-chaperones have been developed aiming to treat cancer. In this review, we summarized HSP70-related cancer signaling pathways and corresponding key proteins regulated by the family of HSP70s. In addition, we also summarized various treatment approaches and progress of anti-tumor therapy based on targeting HSP70 family proteins.

Investigating Salinity Effects in Brackish Aquaponics Systems: Evidencing the Co-Cultivation of the Halophyte Crithmum maritimum with the Euryhaline Sparus aurata

The possibility of simultaneous production of halophyte and euryhaline fish creates huge interest in both commercial aquaponics systems and in research. The aim of the present study was to investigate the effect of three different salinities (8, 14, and 20 ppt) on the growth performance and survival rate of sea bream (Sparus aurata) and rock samphire (Crithmum maritimum) in an experimental brackish aquaponic system. Furthermore, induction of heat shock proteins (Hsps) and phosphorylation of mitogen-activated protein kinases (MAPKs) were assessed through the sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot analysis. A total number of 234 sea bream individuals were divided into nine autonomous aquaponic systems. The experiment lasted 45 days. In total, 54 individuals of rock samphire were used and were distributed into groups of six individuals per hydroponic tank using the raft method. Water quality showed stable fluctuation throughout the experiment, strongly supporting fish and plant growth performance and survival in both treatments. The results show that better growth performance for both sea bream and rock samphire (height increase) was evident in salinity 8 ppt compared to salinities 14 ppt and 20 ppt. Minimal or mild histopathological alterations were detected in gills, midgut, and liver for all three salinity groups. Exposure to different salinities modified Hsp60 and MAPKs expression in a tissue- and time-specific manner. During exposure to 8 ppt, constant Hsp60 levels and phosphorylation of MAPKs at 15 days may indicate a prominent protective role in the gills. The results show that sea bream and rock samphire can be used in brackish aquaponics systems with satisfactory growth performances, thus allowing for a range of commercial applications generating interest in their production.