High Expression Of HSP90 Research Articles

Cloning and functional analysis of heat shock protein Hsp70 from Sclerotinia sclerotiorum

To clarify the roles of the heat shock protein gene Hsp70 in the sclerotial formation and pathogenicity of Sclerotinia sclerotiorum, we employed reverse transcription PCR (RT-PCR) to clone Hsp70 from S. sclerotiorum and performed sequence analysis. Quantitative real-time PCR (qRT-PCR) was employed to determine the relative expression levels of Hsp70 at different growth stages and under the stress induced by cyclic adenosine monophosphate (cAMP) and low and high temperatures. The thermal stability of Hsp70 was measured. The Agrobacterium-mediated method was employed to construct the Hsp70-silenced strain. The pathogenicity and fungicide resistance of strains were tested by inoculation in detached rapeseed leaves and cultivation in the media containing procymidone and thiophanate-methyl, respectively. The results showed that the cloned Hsp70 had a total length of 1 890 bp and close relationship with the Hsp70 gene of Ciborinia. Hsp70 showcased the highest expression level in sclerotia, which was more than 30 times higher than that in hyphae. The cAMP stress significantly induced the expression of Hsp70. The expression level of Hsp70 showed an increasing-decreasing-increasing trend at 40 ℃ and no significant change at 4 ℃. Recombinant strain with high expression of Hsp70 showed good thermal stability. The Hsp70-silenced transformant did not form sclerotia, with decreased pathogenicity and fungicide resistance. This study reveals that Hsp70 plays an important role in the sclerotial formation and stress resistance of S. sclerotium, providing reference for further in-depth research on the biological roles of Hsp70 in S. sclerotium.

Hsp90 Promotes Gastric Cancer Cell Metastasis and Stemness by Regulating the Regional Distribution of Glycolysis-Related Metabolic Enzymes in the Cytoplasm.

Heat-shock protein 90 (Hsp90) plays a crucial role in tumorigenesis and tumor progression; however, its mechanism of action in gastric cancer (GC) remains unclear. Here, the role of Hsp90 in GC metabolism is the focus of this research. High expression of Hsp90 in GC tissues can interact with glycolysis, collectively affecting prognosis in clinical samples. Both in vitro and in vivo experiments demonstrate that Hsp90 is able to regulate the migration and stemness properties of GC cells. Metabolic phenotype analyses indicate that Hsp90 influences glycolytic metabolism. Mechanistically, Hsp90 interacts with glycolysis-related enzymes, forming multienzyme complexes to enhance glycolysis efficiency and yield. Additionally, Hsp90 binds to cytoskeleton-related proteins, regulating the regional distribution of glycolytic enzymes at the cell margin and lamellar pseudopods. This effect could lead to a local increase in efficient energy supply from glycolysis, further promoting epithelial-mesenchymal transition (EMT) and metastasis. In summary, Hsp90, through its interaction with metabolic enzymes related to glycolysis, forms multi-enzyme complexes and regulates regional distribution of glycolysis by dynamic cytoskeletal adjustments, thereby promoting the migration and stemness of GC cells. These conclusions also support the potential for a combined targeted approach involving Hsp90, glycolysis, and the cytoskeleton in clinical therapy.

Correlation of Serum Chemokine (C-C Motif) Ligand 21 and Heat Shock Protein 90 with Preeclampsia.

Objective The study aimed to explore the correlation of serum chemokine (C-C motif) ligand 21 (CCL21) and heat shock protein 90 (Hsp90) with preeclampsia (PE). Methods Between June 2021 and June 2022, 50 pregnant women with PE were included in the PE group, and 50 healthy pregnant women were included in the control group. The serum levels of CCL21 and Hsp90 were compared between the two groups. Results PE patients showed significantly higher levels of CCL21 and Hsp90 than healthy pregnant women (P < 0.05). Correlation analysis showed a positive correlation between CCL21 and Hsp90 levels (r > 0, (P < 0.05)). Binary logistic regression analysis suggested that high expression of CCL21 and Hsp90 were influencing factors for PE (OR >1, (P < 0.05)). The area under the receiver operating characteristic (AUC) curves of Hsp90 and CCL21 levels for predicting PE were 0.895 and 0.864, respectively, suggesting a good predictive value. Conclusion Serum CCL21 and Hsp90 show great potential as disease markers for PE prediction. Further trials are, however, required prior to clinical promotion.

Chronic Administration of COVID-19 Drugs Fluvoxamine and Lopinavir Shortens Action Potential Duration by Inhibiting the Human Ether-à-go-go-Related Gene and Cav1.2.

Background: Old drugs for new indications in the novel coronavirus disease of 2019 (COVID-19) pandemic have raised concerns regarding cardiotoxicity, especially the development of drug-induced QT prolongation. The acute blocking of the cardiac hERG potassium channel is conventionally thought to be the primary mechanism of QT prolongation induced by COVID-19 drugs fluvoxamine (FLV) and lopinavir (LPV). The chronic impact of these medications on the hERG expression has yet to be determined. Methods: To investigate the effect of long-term incubation of FLV and LPV on the hERG channel, we used electrophysiological assays and molecular experiments, such as Western blot, RT-qPCR, and immunofluorescence, in HEK-293 cells stably expressing hERG and human-induced pluripotent stem cell–derived cardiomyocytes (hiPSC-CMs). Results: Compared to the acute effects, chronic incubation for FLV and LPV generated much lower half-maximal inhibitory concentration (IC50) values, along with a left-shifted activation curve and retarded channel activation. Inconsistent with the reduction in current, we unexpectedly found that the chronic effects of drugs promoted the maturation of hERG proteins, accompanied by the high expression of Hsp70 and low expression of Hsp90. Targeting Hsp70 using siRNA was able to reverse the effects of these drugs on hERG proteins. In addition, FLV and LPV resulted in a significant reduction of APD90 and triggered the early after-depolarizations (EADs), as well as inhibited the protein level of the L-type voltage–operated calcium channel (L-VOCC) in hiPSC-CMs. Conclusion: Chronic incubation with FLV and LPV produced more severe channel-blocking effects and contributed to altered channel gating and shortened action potential duration by inhibiting hERG and Cav1.2.

HSP70 and HSP90 in Cancer: Cytosolic, Endoplasmic Reticulum and Mitochondrial Chaperones of Tumorigenesis.

HSP70 and HSP90 are two powerful chaperone machineries involved in survival and proliferation of tumor cells. Residing in various cellular compartments, HSP70 and HSP90 perform specific functions. Concurrently, HSP70 and HSP90 homologs may also translocate from their primary site under various stress conditions. Herein, we address the current literature on the role of HSP70 and HSP90 chaperone networks in cancer. The goal is to provide a comprehensive review on the functions of cytosolic, mitochondrial and endoplasmic reticulum HSP70 and HSP90 homologs in cancer. Given that high expression of HSP70 and HSP90 enhances tumor development and associates with tumor aggressiveness, further understanding of HSP70 and HSP90 chaperone networks may provide clues for the discoveries of novel anti-cancer therapies.

Expression of Heat Shock Protein 70 (Hsp-70) in Children’s Acute Lymphoblastic Leukemia

Heat shock protein (Hsp) 70 is one of the most common Hsp proteins found in humans. Like other types of Hsp, Hsp-70 acts as a chaperone protein in the process of protein synthesis. Its important role in protein synthesis means that these proteins are supposed to be inside the cell and cannot be examined outside the cell. In its development, Hsp-70 turned out to be examined outside of cells due to an increase in Hsp-70 caused by stress on cells which were caused by environmental stress conditions (heat shock, cold shock, heavy metal, oxygen-free radicals), inflammatory and infectious processes, ischemia, tissue damage, aging, and oncogenesis. Cancer studies have found high expression of Hsp-70 in solid tumors and hematological malignancies including ALL. High expression of Hsp-70 is said to have adverse implications on the outcome of ALL and therefore Hsp-70 is auspicious as a therapeutic target in the pediatric treatment of ALL. This review aims to elucidate the role of Hsp-70 expression in pediatric ALL patients.

Mechanisms of heat and hypoxia defense in hard clam: Insights from transcriptome analysis

With global changes and anthropogenic activities, heat and hypoxia have become frequent stressors in aquatic environments. However, the molecular defense mechanisms of aquatic organisms, especially bivalves, against the synergistic effects of heat and hypoxia have not been fully elucidated. In this study, we analyzed transcriptomic data of the gills of Mercenaria mercenaria, an economically and ecologically important bivalve, under several stress challenges: high temperature (heat), low dissolved oxygen (hypoxia), and heat plus hypoxia. GO and KEGG enrichment analyses were performed on the differentially expressed gene sets of heat-induced genes under various dissolved oxygen (DO) solubilities, hypoxia-induced genes under different (normal and high) temperatures and common heat plus hypoxia stress-induced genes. We found that protein-folding item was significantly enriched in common differentially expressed heat-induced genes comparisons, with a high expression of molecular chaperones HSP90 and TCP1. Moreover, microtubule-related GO terms were significantly enriched in differentially expressed hypoxia-induced genes under different temperatures, and kinesin and dynein-related genes were commonly upregulated in the hypoxia challenge groups. Protein processing in endoplasmic reticulum and ubiquitin-mediated proteolysis were the most significantly enriched pathways in common heat plus hypoxia stress-induced genes. Furthermore, correctly-folding related genes, luminal chaperones, and ER-associated degradation related genes were upregulated under heat plus hypoxia stresses. Altogether, our results provided new insights on understanding the molecular defense mechanisms against heat and hypoxia stress in hard clams.

Phytosulfokine \u03b1 (PSK\u03b1) delays senescence and reinforces SUMO1/SUMO E3 ligase SIZ1 signaling pathway in cut rose flowers (Rosa hybrida cv. Angelina)

Roses are widely used as cut flowers worldwide. Petal senescence confines the decorative quality of cut rose flowers, an impressively considerable economic loss. Herein, we investigated the SUMO1/SUMO E3 ligase SIZ1 signaling pathway during bud opening, and petal senescence of cut rose flowers. Our results exhibited that the higher expression of SUMO1 and SUMO E3 ligase SIZ1 during bud opening was accompanied by lower endogenous H2O2 accumulation arising from higher expression and activities of SOD, CAT, APX, and GR, promoting proline accumulation by increasing P5CS expression and activity and enhancing GABA accumulation by increasing GAD expression and activity. In harvested flowers, lower expressions of SUMO1 and SUMO E3 ligase SIZ1 during petal senescence were associated with higher endogenous H2O2 accumulation due to lower expression and activities of SOD, CAT, APX, and GR. Therefore, promoting the activity of the GABA shunt pathway as realized by higher expression and activities of GABA-T and SSADH accompanied by increasing OAT expression and activity for sufficiently supply proline in rose flowers during petal senescence might serve as an endogenous antisenescence mechanism for slowing down petals senescence by avoiding endogenous H2O2 accumulation. Following phytosulfokine α (PSKα) application, postponing petal senescence in cut rose flowers could be ascribed to higher expression of SUMO1 and SUMO E3 ligase SIZ1 accompanied by higher expression and activities of SOD, CAT, APX, and GR, higher activity of GABA shunt pathway as realized by higher expression and activities of GAD, GABA-T, and SSADH, higher expression and activities of P5CS and OAT for supplying proline and higher expression of HSP70 and HSP90. Therefore, our results highlight the potential of the PSKα as a promising antisenescence signaling peptide in the floriculture industry for postponing senescence and extending the vase life of cut rose flowers.

ERK1/2 mitogen-activated protein kinase mediates downregulation of intestinal tight junction proteins in heat stress-induced IBD model in pig.

In many mammalian species, including pigs, heat stress (HS) detrimentally leads to epithelium damage and increases intestinal permeability. However, the underlying molecular mechanisms are not thoroughly investigated yet. This study aimed to examine the RIP1/RIP3-ERK1/2 signaling pathway that regulates the expression of tight junction proteins in HS-treated pigs. In in vitro cultured intestinal porcine epithelial cells (IPEC-J2), HS induced the expression of tight junction proteins, ZO-1, claudin-1, and claudin-4, that are regulated by the ERK1/2-MAPK signaling pathway. Further, high expression of HSP70 in IPEC-J2 cells induced a significant decrease in receptor-interacting protein 1/3 (RIP1/3), phosphorylated ERK, and tight junction protein claudin-1 (P<0.05). Necrostatin-1 (A selective inhibitor of RIPK1) suppressed the upregulation of phosphorylated ERK1/2 induced by HS, indicating that the RIP1/RIP3 regulates ERK1/2 phosphorylation in IPEC-J2 under heat stress. In addition, HS significantly damaged the intestinal morphology characterized by reduction of villus length and crypt depth in in vivo porcine model. Moreover, the expression of tight junction, ZO-1, and claudin-4 were downregulated, whereas phosphorylated p38 and ERK1/2 were upregulated in the duodenum of heat-stressed pigs. Interestingly, a decrease in ZO-1 and claudin-1 was observed in the colon, where phosphorylated ERK1/2 was similar to that in the duodenum. Our results demonstrate that RIP1/RIP3-ERK1/2 signaling pathway regulates the expression of tight junction proteins in HS-pigs. This finding further advances the intestinal barrier function's underlying mechanisms associated with signaling regulation.

Long Noncoding RNA RP11-115N4.1 Promotes Inflammatory Responses by Interacting With HNRNPH3 and Enhancing the Transcription of HSP70 in Unexplained Recurrent Spontaneous Abortion.

BackgroundUnexplained recurrent spontaneous abortion (URSA) is a common pregnancy complication and the etiology is unknown. URSA-associated lncRNAs are expected to be potential biomarkers for diagnosis, and might be related to the disease pathogenesis.ObjectiveTo investigate differential lncRNAs in peripheral blood of non-pregnant URSA patients and matched healthy control women and to explore the possible mechanism of differential lncRNAs leading to URSA.MethodsWe profiled lncRNAs expression in peripheral blood from 5 non-pregnant URSA patients and 5 matched healthy control women by lncRNA microarray analysis. Functions of URSA-associated lncRNAs were further investigated in vitro.ResultsRP11-115N4.1 was identified as the most differentially expressed lncRNA which was highly upregulated in peripheral blood of non-pregnant URSA patients (P = 3.63E-07, Fold change = 2.96), and this dysregulation was further validated in approximately 26.67% additional patients (4/15). RP11-115N4.1 expression was detected in both lymphocytes and monocytes of human peripheral blood, and in vitro overexpression of RP11-115N4.1 decreased cell proliferation in K562 cells significantly. Furthermore, heat-shock HSP70 genes (HSPA1A and HSPA1B) were found to be significantly upregulated upon RP11-115N4.1 overexpression by transcriptome analysis (HSPA1A (P = 4.39E-08, Fold change = 4.17), HSPA1B (P = 2.26E-06, Fold change = 2.99)). RNA pull down and RNA immunoprecipitation assay (RIP) analysis demonstrated that RP11-115N4.1 bound to HNRNPH3 protein directly, which in turn activate heat-shock proteins (HSP70) analyzed by protein-protein interaction and HNRNPH3 knockdown assays. Most importantly, the high expression of HSP70 was also verified in the serum of URSA patients and the supernatant of K562 cells with RP11-115N4.1 activation, and HSP70 in supernatant can exacerbate inflammatory responses in monocytes by inducing IL-6, IL-1β, and TNF-α and inhibit the migration of trophoblast cells, which might associate with URSA.ConclusionOur results demonstrated that the activation of RP11-115N4.1 can significantly increase the protein level of HSP70 via binding to HNRNPH3, which may modulate the immune responses and related to URSA. Moreover, RP11-115N4.1 may be a novel etiological biomarker and a new therapeutic target for URSA.

Dietary inclusion of the red seaweed Asparagopsis taxiformis boosts production, stimulates immune response and modulates gut microbiota in Atlantic salmon, Salmo salar

Salmonids represent US$23 billion of global aquaculture value, yet Atlantic salmon farms lose approximately 10% of their production to diseases and parasites every year. New approaches to minimise such losses are urgently needed because current treatments (e.g. antibiotics) have environmental and human health impacts with increasing sea temperatures predicted to further exacerbate the impacts of disease. Immunostimulants that boost fish resistance to disease without negative environmental or human health impacts are currently being assessed. Seaweeds and their extracts are used as immunostimulants for land animals and are increasingly being investigated for use in finfish aquaculture, including for Atlantic salmon. Here we show that when the red seaweed Asparagopsis taxiformis and its extract were incorporated in Atlantic salmon feed, fish growth rates were enhanced up to 33%, feed intake was enhanced up to 13%, FCR were reduced, and innate immune responses were enhanced up to 58% compared to fish fed unsupplemented control diets over 4 weeks. Overall, fish fed the methanolic extract of A. taxiformis (at an inclusion of ~1% on a dry weight basis, D:D of feed) had the best combination of enhanced growth rate, feed intake and immune response. Fish fed the immunostimulant lipopolysaccharide (LPS) derived from Escherichia coli had the highest innate immune response in our trial, however LPS had no enhanced effect on growth or feed intake. Additionally, we provide evidence that the seaweed and LPS supplements modulated the expression of immune and stress-related genes in both the liver and head kidneys. More specifically, the fish fed the supplemented diets showed increased expression of the HSP70 gene in both their liver and head kidney after 2 weeks of treatment. At 4 weeks high HSP70 and lysozyme gene expression was observed in the fish fed the two seaweed methanolic extract diets. The seaweed diets also enhanced the diversity of bacterial communities within the hindgut of Atlantic salmon while the LPS treatment appeared to have the opposite effect. Whole A. taxiformis or its methanolic extracts could therefore be used as functional feed ingredients that boost the immune response and enhance the growth rate of Atlantic salmon without affecting feed efficiencies.

Higher acclimation temperature affects growth of rohu (Labeorohita) through suppression of GH and IGFs genes expression actuating stress response.

Water temperature alone can affect the growth, metabolic rates and physiological responses of aquatic organisms. Our earlier study reported that higher temperature affects cellular and hemato-biochemical responses in rohu, Labeo rohita. In this backdrop, the present study assessed the effect of higher acclimation temperature on the regulatory mechanisms of growth and stress responses of juvenile L. rohita acclimatized in three temperature conditions (30°C, 33°C, and 36°C) for a period of 30 days. The relative expression of genes for growth hormone (GH), insulin-like growth factors (IGF-1 and IGF-2) and heat shock proteins (hsp70 and hsp90) were measured by real-time quantitative PCR. The results revealed that the highest acclimation temperature (36°C) significantly decreased the weight gain (WG) and specific growth rate (SGR), and increased the feed conversion ratio (FCR) compared to 30°C (control), while increased WG, SGR and lowered FCR were observed in fish reared at the intermediate temperature (33°C) compared to 30°C. Similarly, the GH gene expression in the pituitary was significantly decreased and increased at 36°C and 33°C, respectively as compared to 30°C. A significantly lower expression of IGF-1 and IGF-2, and higher expression of hsp70 and hsp90 were observed in the liver of fish at 36°C. The results of the present study indicate that although slightly elevated temperature promotes the growth of juvenile L. rohita, the higher acclimation temperature may induce stress response and impair growth performance by suppressing GH/IGF system.

Differential expression of angiogenesis markers HSP70, HSP90, VEGF and pERK1/2 in both components of dedifferentiated chondrosarcomas

Dedifferentiated chondrosarcomas (DDCS) are highly malignant bimorphic mesenchymal tumors with poor outcome and limited treatment options. Genes and proteins involved in angiogenesis play an important role in the development of invasion and metastasis. Immunohistochemical stains targeting HSP70, pERK1/2 and VEGFA were applied to a TMA containing 29 DDCS cases representing both tumor components. Higher expression of HSP70 and pERK1/2 was noted in the dedifferentiated component. RNA sequencing performed in 8 paired cases of DDCS comparing well differentiated and dedifferentiated components, showed higher expression of several HSP70 family members and HSP90 in the dedifferentiated component. Furthermore, high mobility group AT-hook 2 (HMAG2) and SET nuclear proto-oncogene demonstrated higher expression in the dedifferentiated component. Thus, the well differentiated and dedifferentiated components of DDCS are different, histologically and transcriptomically. The dedifferentiated component of DDCS shows higher expression of markers that are associated with malignant behavior. Some of these may represent future treatment targets.

Potential Role of Hsp70 and Activated NK Cells for Prediction of Prognosis in Glioblastoma Patients.

Despite rapid progress in the treatment of many cancers, glioblastoma remains a devastating disease with dismal prognosis. The aim of this study was to identify chaperone- and immune-related biomarkers to improve prediction of outcome in glioblastoma. Depending on its intra- or extracellular localization the major stress-inducible heat shock protein 70 (Hsp70) fulfills different tasks. In the cytosol Hsp70 interferes with pro-apoptotic signaling pathways and thereby protects tumor cells from programmed cell death. Extracellular Hsp70 together with pro-inflammatory cytokines are reported to stimulate the expression of activatory NK cell receptors, recognizing highly aggressive human tumor cells that present Hsp70 on their cell surface. Therefore, intra-, extracellular and membrane-bound Hsp70 levels were assessed in gliomas together with activatory NK cell receptors. All gliomas were found to be membrane Hsp70-positive and high grade gliomas more frequently show an overexpression of Hsp70 in the nucleus and cytosol. Significantly elevated extracellular Hsp70 levels are detected in glioblastomas with large necrotic areas. Overall survival (OS) is more favorable in patients with low Hsp70 serum levels indicating that a high Hsp70 expression is associated with an unfavorable prognosis. The data provide a first hint that elevated frequencies of activated NK cells at diagnosis might be associated with a better clinical outcome.

Modulated Electro-Hyperthermia Facilitates NK-Cell Infiltration and Growth Arrest of Human A2058 Melanoma in a Xenograft Model.

Modulated electro-hyperthermia (mEHT), induced by 13.56 MHz radiofrequency, has been demonstrated both in preclinical and clinical studies to efficiently induce tumor damage and complement other treatment modalities. Here, we used a mouse xenograft model of human melanoma (A2058) to test mEHT (~42°C) both alone and combined with NK-cell immunotherapy. A single 30 min shot of mEHT resulted in significant tumor damage due to induced stress, marked by high hsp70 expression followed by significant upregulation of cleaved/activated caspase-3 and p53. When mEHT was combined with either primary human NK cells or the IL-2 independent NK-92MI cell line injected subcutaneously, the accumulation of NK cells was observed at the mEHT pretreated melanoma nodules but not at the untreated controls. mEHT induced the upregulation of the chemoattractant CXCL11 and increased the expression of the matrix metalloproteinase MMP2 which could account for the NK-cell attraction into the treated melanoma. In conclusion, mEHT monotherapy of melanoma xenograft tumors induced irreversible heat and cell stress leading to caspase dependent apoptosis to be driven by p53. mEHT could support the intratumoral attraction of distantly injected NK-cells, contributed by CXCL11 and MMP2 upregulation, resulting in an additive tumor destruction and growth inhibition. Therefore, mEHT may offer itself as a good partner for immunotherapy.

Cell proliferation and wound healing effects of LEDs in the olive flounder, Paralichthys olivaceus

Physical wounds in fish can devastate bodily homeostasis, increase the likelihood of secondary infections, and cause massive economic damage. Due to the importance of skin as a primary physical barrier, the strategy of promptly healing physical wounds to reduce further negative consequences has attracted a great deal of attention. The purpose of this study was to accelerate physical wound healing in the flounder using LED technology, which can be applied to thousands of fish at the same time at a reasonable economic cost. The fish epithelial cell line (Epithelioma papulosum cyprini; EPC) was used to verify the proliferative effects of LED irradiation at wavelengths of 465, 520, and 640 nm at 0, 10, 20, 40, and 80 μ·mol·m−2·s−1. Also, desquamated wounds were made to flounder with a razor, and the wounds were exposed to LED light at 520 and 640 nm, as well as ambient light as a control, in 12-h light/ dark cycles for seven or 28 days. The results showed that 520 and 640 nm LED treatment stimulated EPC cell proliferation, while the no-LED and 465 nm LED conditions had no effect. After the injuries, the mortality rate of fish exposed to 520 nm LED was markedly lower than those of the control (ambient light) and 640 nm-treated group. Also, the highest re-generated epithelium thickness (1-day post-injury (dpi)) and thicker collagen and connective tissue (7 dpi) were observed in the 520 nm group, along with high expression of IL-1, HSP70, TNF-α, and MMP-13. Likewise, significantly faster wound closure rates were observed with ample recovered muscle layers in the 520 nm treatment group at 21 and 28 dpi. These results show that 520 nm LED treatment is a very effective alternative method of aiding recovery from physical injuries, and is an eco-friendly method that can easily be applied in aquaculture.

HSP90 expression and early recurrence in gastroenteropancreatic neuroendocrine tumors: Potential for a novel therapeutic target

BackgroundHeat shock protein (HSP)-90 promotes tumor growth and is overexpressed in many malignancies. HSP90 expression profile and its potential as a therapeutic target in primary and metastatic neuroendocrine tumors (NETs) are not known. MethodsHSP90 cytoplasmic expression and Ki-67 index were re-reviewed and scored by a pathologist blinded to all other clinicopathologic variables for patients who underwent resection of primary and metastatic gastroenteropancreatic (GEP) neuroendocrine tumors at a single institution (2000–2013). Primary outcome was recurrence-free survival (RFS). ResultsOf 263 tumors reviewed, 73% (n = 191) were primary GEP NETs, and 12% (n = 31) were NET liver metastases. Of the primary GEP-NETs, mean age was 56 years, 42% were male; 53% (n = 103) were pancreatic and 23% (n = 44) were small bowel. HSP90 expression was high in 34% (n = 64) and low in 66% (n = 127). Compared to low expression, high HSP90 was associated with advanced T-stage (T3/T4) (47 vs 27%; p = 0.02). Among patients who underwent curative-intent resections for primary, non-metastatic NETs (n = 145), high HSP90 was independently associated with worse RFS (HR 5.09, 95% CI 1.65–15.74; p = 0.005), after accounting for positive margin, LN involvement, increased tumor size, site of primary tumor, and Ki-67. When assessing NET liver metastases, 13% (n = 4) had high HSP90 expression and 87% (n = 26) had low expression. Patients with liver metastases with high HSP90 tended to have worse 1- and 3-year progression-free survival (25%, 25%) compared to those with low HSP90 (69%, 49%; p = 0.059). ConclusionHSP90 exhibits differential expression in resected GEP-NETs and liver metastases. High cytoplasmic expression is associated with early disease recurrence, even after accounting for other adverse pathologic factors. HSP90 inhibition may be a potential therapeutic target for neuroendocrine tumors.

Prebiotic agavin in juvenile totoaba, Totoaba macdonaldi diets, to relieve soybean meal‐induced enteritis: Growth performance, gut histology and microbiota

The prebiotic effect of the agave plant polysaccharides (agavin) was evaluated to improve enteritis caused by soybean meal (SBM) in juveniles of Totoaba macdonaldi (60.8 ± 2.3 g). Four isoproteic and isolipidic diets were formulated: an FM‐based formulation as reference (RD), a blend of FM and PBM (FM‐PBM; 1:2 as previously shown better performance than FM), the blend combined with 24% SBM (SBM‐B) and, lastly, the latter combination with 2% agavin (SBM‐B + A). After 44 days, the SBM‐B + A treatment resulted in a definite enteritis improvement, besides a significantly higher final growth (207.1 ± 3.8 vs. 195 ± 3.4) compared to the rest of the treatments. According to the microbiota intestine structure, it can be revealed that agavin resulted in enrichment of the Sinorhizobium genera, characterized by nitrogen fixation. However, at a species level, Clostridium botyricum was the more prominent in PBM‐based feeds, known as beneficial for the suppression of inflammatory response. In contrast, Enterococcus cecorum was identified in the gut of fish fed SBM‐based feeds, intensified in agavin‐supplemented feeds. Higher expression of hsp70 and aqp8 was registered in FM‐PBM. It is concluded that agavin provides a clear benefit when added to diets. The implications of FM substitution are discussed.

Abstract 6396: Sarcoma treatment using HSP90 inhibitors in combinatorial synergy with doxorubicin

Abstract Synovial sarcoma, a rare muscle cancer, has few targeted therapies, and is most commonly treated using doxorubicin (DOX). Even as a primary treatment agent, successful treatment and remission rates are low, and the significant cardiotoxicity of DOX limits dosage options. Synovial sarcoma, and other sarcomas, have a high expression of HSP90, a protein that acts as a chaperone molecule to stabilize and help fold other proteins within the cellular matrix. Under high stress situations, HSP90 is upregulated and acts to stabilize other more thermally labile proteins, including many oncogenes. Using FDA approved drug panels, it was determined that HSP90 is a suitable target for treatment. We look to use a novel HSP90 inhibitor (HS10) compound, to target the high expression of HSP90 within synovial sarcoma and osteosarcoma in combination with the chemotherapy standard DOX to determine if a synergistic effect is observed. This synergistic effect would allow for an overall decreased dose to induce effective cancer cell targeting, reducing overall systemic toxicity within patients. Our results indicate such synergism does occur between DOX and HS10, at a dosage of 5-25 nM DOX and 1000 nM HS10, reducing cell survival to 37-68%, dependent on cell line. DOX dosage alone reduced cell survival to 74% in the most effective case, with no reduction in the least effective case, at a dose of 100 nM. This dose is four times stronger than the highest DOX synergistic dose. It can be concluded, then, that synergistic combination does exist between DOX and HS10, and dosage of DOX can be reduced with more effective targeting for synovial and osteosarcoma cell lines. Protein expression was evaluated using immunoblotting to isolate protein fractions within the cytoplasm and cell membrane, showing high expression within the cytoplasm. Isolated blood plasma from mice with and without tumors was analyzed through ELISA, showing elevated HSP90 presence in mice with tumors (236.4±190pg) vs mice without tumors (6.5±3.5pg). This confirmation of elevated HSP90 expression within sarcoma patients suggests targeting using HS10 to induce synergy to be a viable approach for advanced sarcoma treatment. Citation Format: Matt Kirkham, Sarah Luelling, Kaniz Fatema, Jared Barrott. Sarcoma treatment using HSP90 inhibitors in combinatorial synergy with doxorubicin [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6396.

Extreme climate scenario and parasitism affect the Amazonian fish Colossoma macropomum

Global warming caused by greenhouse gases accumulation, in particular carbon dioxide, is the major current environmental challenge, as it will affect all life forms over the next decades. Aquaculture, a fast growing food production sector, is already facing the effects of global warming. The fish immune system is expected to be especially affected by increased temperature and carbon dioxide, mainly when associated to infectious diseases outbreaks. Here, we experimentally investigated the associated effects of an extreme climate scenario and two levels of monogenean parasitism on the hematological and immunological response of an important Amazon fish for continental aquaculture: Colossoma macropomum. Individuals of C. macropomum with low and high levels of parasitism were exposed to current and extreme climate scenarios (4.5 °C and 900 ppm CO2 above current levels). We characterized their hematological profile using classical methods, and their immune-related gills gene expression (HSP70, IL-1β and IL-10) using quantitative real-time polymerase chain reaction (qPCR). After 7 days of exposure, we observed that exposure to extreme climate scenario caused rapid increase of parasitism intensity and likely acute inflammation, indicated by the higher expression of HSP70 and IL-1β. The IL-10 gene was downregulated in both groups exposed to extreme climate scenario, contrasting with animals exposed to current scenario. Thus, in the current scenario, the parasitized tambaqui showed a response to the tissue damage, which was not observed in the animals exposed to the extreme scenario.