Heat Shock Protein 90α Research Articles

Potential predictive value of plasma heat shock protein 90α in lung cancer.

ObjectiveHeat shock protein 90α (HSP90α) is associated with cancer development, progression, and metastasis. This study assessed the relationships of plasma HSP90α levels with treatment efficacy and prognosis in lung cancer.MethodsIn this retrospective cross-sectional study, 231 patients with lung cancer were enrolled from 1 September 2016 to 31 December 2019. HSP90α levels were measured before and after treatment, and their relationships with outcomes were assessed.ResultsPatients with elevated HSP90α levels before treatment had a better overall response rate (ORR, 44.1% vs. 30.6%), whereas the disease control rate did not differ between patients with elevated and normal HSP90α levels (81% vs. 78.5%). Median progression-free survival (PFS) was 6.9 months in patients with elevated baseline HSP90α levels, versus 9 months in patients with normal HSP90α levels, whereas the median overall survival (OS) times in these groups were 12 and 14.1 months, respectively. Concerning HSP90α levels after treatment, ORR (20% vs. 47.1%) and DCR (67.3% vs. 90.9%) were lower in patients with increased HSP90α levels, and PFS and OS were also significantly different between the groups.ConclusionsHSP90α levels before and after treatment were associated with treatment response and patient prognosis in lung cancer.

Predictive Modeling of MAFLD Based on Hsp90α and the Therapeutic Application of Teprenone in a Diet-Induced Mouse Model

Background and AimsThe heat shock protein (Hsp) 90α is induced by stress and regulates inflammation through multiple pathways. Elevated serum Hsp90α had been found in nonalcoholic steatohepatitis (NASH). Geranylgeranylacetone (GGA, also called teprenone) is a terpenoid derivative. It was reported to induce Hsp and alleviate insulin resistance. We aimed to evaluate the Hsp90α as a biomarker in predicting metabolic-associated fatty liver disease (MAFLD) and define the therapeutic effects of geranylgeranylacetone for the disease.MethodsA clinical study was conducted to analyze the elements associated with Hsp90α, and a predictive model of MAFLD was developed based on Hsp90α. The histopathological correlation between Hsp90α and MAFLD was investigated through a diet-induced mouse model. Furthermore, GGA was applied to the mouse model.ResultsSerum Hsp90α was increased in patients with MAFLD. A positive linear relationship was found between age, glycosylated hemoglobin (HbA1c), MAFLD, and serum Hsp90α. Meanwhile, a negative linear relationship with body mass index (BMI) was found. A model using Hsp90α, BMI, HbA1c, and ALT was established for predicting MAFLD. The area under the receiver operating characteristic (ROC) curves was 0.94 (95% CI 0.909–0.971, p = 0.000). The sensitivity was 84.1%, and the specificity was 93.1%. In vitro experiments, GGA induced Hsp90α in steatosis cells. In the mice model, Hsp90α decreased in the GGA treatment group. Hepatic steatosis, inflammation, insulin resistance, and glucose intolerance were improved in the GGA-treated group. Serum Hsp90α was positively correlated with steatohepatitis activity according to hepatic histopathology.ConclusionsSerum Hsp90α was elevated in MAFLD, and a positive correlation between serum Hsp90α and the grade of activity of steatohepatitis was observed. The model using BMI, HbA1c, and alanine aminotransferase (ALT) had a good value to predict MAFLD. The findings also revealed the effectiveness of GGA in the treatment of MAFLD.

The Role of AKR1B10 in Physiology and Pathophysiology.

AKR1B10 is a human nicotinamide adenine dinucleotide phosphate (NADPH)-dependent reductase belonging to the aldo-keto reductase (AKR) 1B subfamily. It catalyzes the reduction of aldehydes, some ketones and quinones, and interacts with acetyl-CoA carboxylase and heat shock protein 90α. The enzyme is highly expressed in epithelial cells of the stomach and intestine, but down-regulated in gastrointestinal cancers and inflammatory bowel diseases. In contrast, AKR1B10 expression is low in other tissues, where the enzyme is upregulated in cancers, as well as in non-alcoholic fatty liver disease and several skin diseases. In addition, the enzyme’s expression is elevated in cancer cells resistant to clinical anti-cancer drugs. Thus, growing evidence supports AKR1B10 as a potential target for diagnosing and treating these diseases. Herein, we reviewed the literature on the roles of AKR1B10 in a healthy gastrointestinal tract, the development and progression of cancers and acquired chemoresistance, in addition to its gene regulation, functions, and inhibitors.

Oviduct fluid during IVF moderately modulates polyspermy in in vitro-produced goat embryos during the non-breeding season

The present study determined i) the presence of proteins (oviduct-specific glycoprotein, OVGP1; heat shock protein-70A, HSPA1A; heat shock protein-A8, HSPA8; annexin A1, ANXA1; annexin A5, ANXA5; and myosin-9, MYH9) known to be involved in early reproduction in the oviduct fluid (OF) of anestrous goats; and ii) the functional effect of during IVF on polyspermy modulation and embryonic development. In vitro-matured oocytes were co-cultured with spermatozoa (1.0, 2.0, or 4.0 x 106 cells/mL) for 18 h in SOF medium supplemented with 5 μg/mL of heparin, 4 μg/mL gentamicin, and 10% estrus sheep serum (CTRL1, CTRL2, and CTRL4 groups) or the same medium plus 10% OF (OF1, OF2, and OF4 groups) obtained from anestrus goats. The analysis of OF by western blotting confirmed the presence of the six proteins tested for. The increase in sperm concentration had no effect (P > 0.05) on the penetration rate in any group; however, monospermy rate decreased as sperm concentration was increased in both OF and CTRL. Regardless of the concentration used, when data were pooled, OF supplementation improved (P < 0.05) monospermy and tended (P = 0.057) to enhance IVF efficiency. Additionally, IVF efficiency was higher (P < 0.05) in OF1 than in OF4 [60 ± 13 vs 37 ± 5%). The development capacity was not affected (P > 0.05) by the sperm concentration and OF treatment, and the average values were cleavage (72 ± 2.6%), blastocyst (37 ± 3.0%), blastocyst in relation to the cleaved (51 ± 4.8%), hatched (62 ± 1.2%), and number of cells per blastocyst (174 ± 1.8%). In conclusion, the six proteins analyzed are present in the OF of anestrous goats, and the supplementation of this OF during IVF may modulate the polyspermy incidence and enhance IVF efficiency, especially when 1x106 sperm per mL is used.

Relationship between 18F-FDG uptake and heat shock protein 90α expression in colorectal cancer.

We investigated whether heat shock protein 90α (HSP90α) expression is associated with fluorine-18-labeled fluoro-2-deoxy-D-glucose (18F-FDG) uptake and whether 18F-FDG positron emission tomography/computed tomography (PET/CT) can be used to predict the status of HSP90α expression in colorectal cancer (CRC). The medical records and preoperative 18F-FDG PET/CT studies of 51 patients with newly diagnosed CRC who underwent surgical treatment were retrospectively reviewed. The maximum standardized uptake value (SUVmax) of the primary tumor was calculated from the level of 18F-FDG uptake. HSP90α expression was determined by immunohistochemistry. The relationship between SUVmax and HSP90α expression was analyzed. Colorectal cancer with high HSP90α expression had significantly higher SUVmax than CRC with low HSP90α expression (18.88±10.06 vs. 12.38±5.04, P=0.003). There was a significant correlation between HSP90α expression and 18F-FDG uptake (r=0.354, P=0.011). The highest accuracy for determining HSP90α status (68.6%) was obtained with a SUVmax cut-off of 15.4. Maximum SUV was the only predictor of HSP90α expression on multivariate logistic regression analysis (Odds Ratio=5.384, P=0.016). The expression status of HSP90α was significantly related to 18F-FDG uptake in CRC.

Diagnostic and prognostic value of heat shock protein 90α in malignant melanoma.

Malignant melanoma is one of the most common tumours of the skin. Heat shock protein 90α (HSP90α) has been applied in the auxiliary diagnosis of various malignancies, as a tumour marker. This study aims to evaluate diagnostic, therapeutic efficacy and prognostic value of plasma HSP90α levels in malignant melanoma. In this study, higher plasma HSP90α levels and abnormal rates were found in malignant melanoma patients than in healthy controls (92.63 vs. 51.84 ng/mL; P < 0.001 and 68.30 vs. 8.30%; P < 0.001). Plasma HSP90α levels were higher with Breslow thickness >4 mm, a high Clark level (IV + V), abnormal serum lactate dehydrogenase (LDH), distant metastases occurrence and Ki-67≥30% (P < 0.05). The area under the curves (AUCs) of HSP90α was greater than LDH in the training (0.847 vs. 0.677) and validation (0.867 vs. 0.672) cohort. Meanwhile, the sensitivity (76.70%) and negative predictive values (78.80%) of HSP90α were higher. Plasma HSP90α levels were significantly reduced in objective response (81.05 vs. 37.26 ng/mL; P = 0.012) and disease control patients (84.16 vs. 47.05 ng/mL; P = 0.002) post-treatment. Patients with normal HSP90α levels had slightly longer progression-free survival (PFS) than those with abnormal levels (8.0 vs. 3.5 months; P = 0.096). Unfortunately, the trend was not statistically significant. In multivariable analysis, immunotherapy was an independent prognostic factor for PFS. Nevertheless, patients with normal HSP90α levels who received chemotherapy(±targeted therapy) without immunotherapy had significantly longer PFS than patients with abnormal levels (6.0 vs. 2.0 months; P = 0.008). Therefore, HSP90α can be used for auxiliary diagnosis and predict the responses to therapy in malignant melanoma patients.

Heat shock protein 90α in nipple discharge as a potential tumor marker for breast cancer.

Heat shock protein 90α (HSP90α) has been confirmed to be upregulated in the blood in various types of tumors and may therefore serve as a potential tumor marker. However, whether HSP90α exists in nipple discharge remains unknown, and its expression and diagnostic value in nipple discharge remain unclear. In this study, the expression of HSP90α, carcinoembryonic antigen (CEA), and cancer antigen 153 in nipple discharge and blood from 128 patients was measured. Receiver operating characteristic curve was used to assess the diagnostic value of HSP90α. Further, its relationship with clinicopathological parameters of patients with breast cancer was analyzed. The results showed that the expression of HSP90α in nipple discharge was significantly higher in patients with breast cancer than in those with benign disease, and its diagnostic value was better than that of CEA. Combination of HSP90α and CEA showed better diagnostic efficacy than HSP90α or CEA alone. Moreover, the expression of HSP90α displayed a stepwise increase from benign lesions, followed by carcinoma in situ to invasive ductal carcinoma. HSP90α was positively correlated with Ki67 expression. However, there was no significant difference in the expression of HSP90α in blood between patients with breast cancer and benign disease. Further, the expression of HSP90α was higher in nipple discharge than in blood. In summary, HSP90α was upregulated in the nipple discharge of patients with breast cancer, and it may be related to the occurrence and progression of breast cancer. HSP90α in nipple discharge may serve as a potential diagnostic marker for breast cancer.

Heat shock protein 90α inhibitor, PU-H71 in combination with DHEA promoting apoptosis in triple-negative breast cancer cell line MDA-MB-231.

Due to the lack of markers (ER, PR, and HER-2/Neu) for the molecular-targeted therapies triple-negative breast cancer (TNBC) is more challenging than other subtypes of breast cancer. Moreover, the conventional chemotherapeutic agents are still the mainstay of most therapeutic protocols and eventually turn into a refractory drug-resistance , hence, more efficient therapeutic regimens are urgently required. The present study aimed to elucidate the effects of PU-H71 combined with DHEA on triple-negative breast cancer cell line MDA-MB-231 and to assess the synergy using the Chou-Talalay method. The combined therapy controlled the expression of an array of antioxidants and metabolizing enzymes, leading to the induction of oxidative stress which in turn induced apoptotic cell death. Our results indicated that the combined treatment with PU-H71 and DHEA exerts a synergistic anti-tumor effect on MDA-MB-231 triple-negative breast cancer cell line.

Heat shock protein 90α increases superoxide generation from neuronal nitric oxide synthases

Neuronal nitric oxide synthase (nNOS) generates superoxide, particularly at sub-optimal l-arginine (l-Arg) substrate concentrations. Heat shock protein 90 (Hsp90) was reported to inhibit superoxide generation from nNOS protein. However, commercially available Hsp90 product from bovine brain tissues with unspecified Hsp90α and Hsp90β contents and an undefined Hsp90 protein oligomeric state was utilized. These two Hsp90s can have opposite effect on superoxide production by NOS. Importantly, emerging evidence indicates that nNOS splice variants are involved in different biological functions by functioning distinctly in redox signaling. In the present work, purified recombinant human Hsp90α, in its native dimeric state, was used in electron paramagnetic resonance (EPR) spin trapping experiments to study the effects of Hsp90α on superoxide generation from nNOS splice variants nNOSμ and nNOSα. Human Hsp90α was found to significantly increase superoxide generation from nNOSμ and nNOSα proteins under l-Arg-depleted conditions and Hsp90α influenced superoxide production by nNOSμ and nNOSα at varying degrees. Imidazole suppressed the spin adduct signal, indicating that superoxide was produced at the heme site of nNOS in the presence of Hsp90α, whereas l-Arg repletion diminished superoxide production by the nNOS-Hsp90α. Moreover, NADPH consumption rate values exhibited a similar trend/difference as a function of Hsp90α and l-Arg. Together, these EPR spin trapping and NADPH oxidation kinetics results demonstrated noticeable Hsp90α-induced increases in superoxide production by nNOS and a distinguishable effect of Hsp90α on nNOSμ and nNOSα proteins.

Heat Shock Protein 90α Provides an Effective and Novel Diagnosis Strategy for Nasopharyngeal Carcinoma.

Nasopharyngeal carcinoma (NPC) is an Epstein-Barr virus (EBV)-associated tumor occurring in southeastern Asia. Due to insidious onset, it is difficult to diagnose NPC from clinical symptoms. Thus, there is an urgent need for non-invasive, high-performance biomarkers to aid the clinical diagnosis of NPC. Heat shock protein 90α (HSP90α) is an important member of the heat shock protein family that significantly increases under stress conditions such as oxidation and tumors. This is the first investigation of the role of Hsp90α in the diagnosis and progress of NPC. Plasma Hsp90α was detected by ELISA in 196 newly diagnosed NPC patients, 76 corresponding post-treatment NPC patients, 230 VCA-IgA-positive normal subjects and 106 healthy controls. (1) The level of Hsp90α in plasma of 196 NPC patients was (212.16 ± 144.32)ng/ml, which was significantly higher than that in VCA-IgA-positive normal subjects (68.12 ± 64.94ng/ml, P < 0.001) and healthy controls (35.87 ± 17.47ng/ml, P < 0.001); (2) the levels of Hsp90α in patients with NPC in the early stage (I + II), stage III and stage IV were significantly different (159.69 ± 117.12pg/ml vs. 195.24 ± 126.38pg/ml vs. 250.85 ± 164.66pg/ml, P = 0.018 and P = 0.029, respectively). The level of Hsp90α in plasma in patients with metastasis of NPC and those without metastasis was significantly different (P < 0.001); (3) Hsp90α is closely related to EBV DNA levels, but not to the VCA-IgA titer and EA-IgA titer; (4) the levels of Hsp90α in plasma of patients with NPC before and after treatment were significantly different (212.16 ± 144.32pg/ml vs. 62.36 ± 34.04pg/ml, P < 0.001); (5) the ROC curves demonstrated that the sensitivity of plasma Hsp90α in distinguishing NPC patients from healthy controls was 74.50% and the specificity was 99.10% (AUC = 0.931, 95% CI 0.903-0.958). The study found that the plasma HSP90α level is closely related to the clinical stage, metastasis and therapeutic effect of NPC. HSP90α may serve as a new biomarker for diagnosis and treatment of NPC.

Apigenin suppresses influenza A virus-induced RIG-I activation and viral replication.

Apigenin is a flavonoid of low toxicity and multiple beneficial bioactivities, including the properties of antitumor, antioxidant, anti-inflammatory, and antiviral activities. However, the effects of Apigenin on influenza virus infection remain poorly understood. Thus, the aim of this study is to investigate the effect of Apigenin on influenza A virus (IAV)-induced inflammation and viral replication. This study demonstrated that Apigenin treatment significantly suppressed IAV-induced upregulation of retinoic acid-inducible gene-I (RIG-I) expression, as well as the production of proinflammatory cytokines and interferons (IFN-β and IFN-λ1). Meanwhile, Apigenin also protected cells from IAV-induced cell death. In addition, Apigenin specifically inhibited the activation of RIG-I signaling via promoting the ubiquitin-mediated degradation of RIG-I, which may cause by the disrupting its interaction with heat shock protein 90α. Interestingly, instead of enhancing viral replication due to the inhibitory effects of Apigenin on the activation of RIG-I and expression of IFNs, Apigenin inhibited IAV replication in vitro. Further study demonstrated that Apigenin inhibited the influenza viral neuraminidase (NA) activity. Thus, Apigenin may serve as a promising supplementary approach for treatment of influenza because it protected cells from IAV-induced cell death and inhibited viral NA activity to suppress viral replication.

SIRT2 suppresses expression of inflammatory factors via Hsp90-glucocorticoid receptor signalling.

SIRT2 is a NAD+‐dependent deacetylase that deacetylates a diverse array of protein substrates and is involved in many cellular processes, including regulation of inflammation. However, its precise role in the inflammatory process has not completely been elucidated. Here, we identify heat‐shock protein 90α (Hsp90α) as novel substrate of SIRT2. Functional investigation suggests that Hsp90 is deacetylated by SIRT2, such that overexpression and knock‐down of SIRT2 altered the acetylation level of Hsp90. This subsequently resulted in disassociation of Hsp90 with glucocorticoid receptor (GR), and translocation of GR to the nucleus. This observation was further confirmed by glucocorticoid response element (GRE)‐driven reporter assay. Nuclear translocation of GR induced by SIRT2 overexpression repressed the expression of inflammatory cytokines, which were even more prominent under lipopolysaccharide (LPS) stimulation. Conversely, SIRT2 knock‐down resulted in the up‐regulation of cytokine expression. Mutation analysis indicated that deacetylation of Hsp90 at K294 is critical for SIRT2‐mediated regulation of cytokine expression. These data suggest that SIRT2 reduces the extent of LPS‐induced inflammation by suppressing the expression of inflammatory factors via SIRT2‐Hsp90‐GR axis.

HSP90α combined with AFP and TK1 improved the diagnostic value for hepatocellular carcinoma.

Aim: We investigated the effect of the combination of heat shock protein 90α (HSP90α), alpha-fetoprotein (AFP) and thymidine kinase 1 (TK1) in the diagnosis of hepatocellular carcinoma (HCC). Methods & results: A total of409 patients with HCC, 101 patients with benign liver disorder and 78 healthy individuals were retrospectively analyzed. HSP90α level was higher in HCC patients than in controls. The expression of HSP90α showed a positive correlation with tumor stage, differentiation, lymph node metastasis and tumor thrombus formation. The combination of HSP90α, AFP and TK1 improved the diagnostic sensitivity (89.24%) and the area under the receiver operating characteristic curve (0.919). Conclusion: The detection of HSP90α, AFP and TK1 is more efficient than a single tumor marker for the diagnosis of HCC.

Cytotoxic necrotizing factor 1 promotes bladder cancer angiogenesis through activating RhoC.

Uropathogenic Escherichia coli (UPEC), a leading cause of urinary tract infections, is associated with prostate and bladder cancers. Cytotoxic necrotizing factor 1 (CNF1) is a key UPEC toxin; however, its role in bladder cancer is unknown. In the present study, we found CNF1induced bladder cancer cells to secrete vascular endothelial growth factor (VEGF) through activating Ras homolog family member C (RhoC), leading to subsequent angiogenesis in the bladder cancer microenvironment. We then investigated that CNF1-mediated RhoC activation modulated the stabilization of hypoxia-inducible factor 1α (HIF1α) to upregulate the VEGF. We demonstrated in vitro that active RhoC increased heat shock factor 1 (HSF1) phosphorylation, which induced the heat shock protein 90α (HSP90α) expression, leading to stabilization of HIF1α. Active RhoC elevated HSP90α, HIF1α, VEGF expression, and angiogenesis in the human bladder cancer xenografts. In addition, HSP90α, HIF1α, and VEGF expression were also found positively correlated with the human bladder cancer development. These results provide a potential mechanism through which UPEC contributes to bladder cancer progression, and may provide potential therapeutic targets for bladder cancer.

In Silico Prediction of the Dissociation Rate Constants of Small Chemical Ligands by 3D-Grid-Based VolSurf Method.

Accumulated evidence suggests that binding kinetic properties—especially dissociation rate constant or drug-target residence time—are crucial factors affecting drug potency. However, quantitative prediction of kinetic properties has always been a challenging task in drug discovery. In this study, the VolSurf method was successfully applied to quantitatively predict the koff values of the small ligands of heat shock protein 90α (HSP90α), adenosine receptor (AR) and p38 mitogen-activated protein kinase (p38 MAPK). The results showed that few VolSurf descriptors can efficiently capture the key ligand surface properties related to dissociation rate; the resulting models demonstrated to be extremely simple, robust and predictive in comparison with available prediction methods. Therefore, it can be concluded that the VolSurf-based prediction method can be widely applied in the ligand-receptor binding kinetics and de novo drug design researches.

Synthesis, evaluation, molecular dynamics simulation and targets identification of novel pyrazole-containing imide derivatives

A new series of novel pyrazole-containing imide derivatives were synthesized and evaluated for their anticancer activities against A-549, Bel7402, and HCT-8 cell lines. Among these compounds A2, A4, A11 and A14 possessed high inhibition activity against A-549 cell lines with IC50 values at 4.91, 3.22, 27.43 and 18.14 μM, respectively, better than that of 5-fluorouracil (IC50=59.27 μM). A2, A4, and A11 also exhibited significant inhibitory activity towards HCT-8 and Bel7402 cell lines. Interestingly, the Heat Shock Protein 90α (Hsp90α, PDB ID: 1UYK) was found to be the potential drug target of these synthesized compounds with the aid of PharmMapper server (http://lilab.ecust.edu.cn/pharmmapper/) and docking module of Schrödinger (Maestro 10.2). Additionally, molecular dynamics simulation was performed out to explore the most likely binding mode of compound A2 with Hsp90α. Communicated by Ramaswamy H. Sarma

Egg White peptide KPHAEVVLR promotes skin fibroblasts migration and mice skin wound healing by stimulating cell membrane Hsp90α secretion

Egg white peptides are well known for their abundant species and biological properties. The aim of this study was to investigate the effect of egg white peptides on skin wound healing. We hydrolyzed the egg white powder using Alcalase and flavourzyme. Our transwell cell migration assay data showed that EWP2, which is one of the four fractions obtained from Sephadex G-15 isolation, promoted human skin fibroblast HSF cells migration. LC–MS/MS analysis results showed EWP2 were consist of 24 high abundant peptides. We randomly chose and synthesized four of these 24 peptides. Transwell assay data displayed that the egg white peptide KPHAEVVLR (KR-9) increased 8.2-fold of HSF cells migration at 200 μM compared with control. 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sμlfophenyl)-2H-tetrazolium (MTS) assay results showed KR-9 did not lead to HSF cells over-proliferation. In vivo animal experiment data proved that KR-9 accelerated 50 % and 136 % mice skin wound healing rate at 50 μM and 100 μM. Furthermore, we explored the mechanism of KR-9. Our western blot and immunofluorescence results showed KR-9 enhanced the protein level of cell membrane heat shock protein 90α (Hsp90α). Thus, we conclude that egg white peptide KR-9 promotes skin fibroblasts migration and mice skin wound healing by stimulating cell membrane Hsp90α secretion.

Abstract P3-01-18: Kinase D interacting substrate 220 (Kidins220) and disease progression of breast cancer, the role of heat shock protein90 (HSP90)

Abstract Background: Kidins220 (Kinase D Interacting Substrate 220) is a transmembrane scaffold protein and was initially found as the substrate of Protein Kinase D. It has several binding partners and plays an important role in neuronal activities and attribute to the occurrence of neuronal disease. Moreover, emerging evidence has revealed the involvement of Kidins220 in regulating cancer development including melanoma, neuroblastoma, and prostate cancer. Our current study investigated the role of Kidins220 in breast cancer and the biological implications on breast cancer cells. Method: A panel of human breast cancer cell lines were used in the biological studies. Kidin220 Knockdown cell model was established using lentivirus anti-Kidins220 shRNA and the knockdown of Kidins220 was verified in gene and protein levels. Cell matrix attachment and cell migration assay were determined using an automated cell analyser (ECIS). The growth of breast cancer cells was evaluated using colorimetric method. Human breast tumours (n=106) together with adjacent background tissues were collected immediately after surgery. Transcription level of Kidins220 and HSPs in the fresh tissues was determined using quantitative transcript analyses. Results: In the clinical cohort of breast cancer, an increase in expression of Kidins220 was found in tumour tissues in comparison with adjacent normal tissues. Patients with poor prognosis (NPI&amp;gt;5.4) had the lowest expression of Kidins220 compared with patients with good and moderate prognosis. Likewise, high grade tumours had markedly reduced level of Kidins220. Furthermore, patients remain disease free had the highest Kidins220 level when compared with those who had breast cancer-related incidence. Kaplan-Meier plot showed high level of Kidins220 was linked to longer survival. Our database also revealed that the levels of expression of Kidins200 were significantly correlated with that of HSP90A (Heat Shock Protein-90A) (r=0.37, p&amp;lt;0.001), but not HSP90B. Moreover, breast cancer cells (MDA MB-231 and MCF-7) after losing Kidins220 by way of knockdown showed an increase in cellular migration and cell matrix adhesion, with little changes in cell growth compared with control cells. In these cell based models, HSP90 inhibitor was found to affect Kidins220 mediated changes in matrix adhesion. It was also noteworthy that cells with Kidins220 knockdown had dramatically reduced expression of HSP90, particularly HSP90A. Conclusion: Kidins220 may act as a tumour suppressor in regulating the disease progression of breast cancer and has a correlation with the survival of patients with breast cancer. Kidins220 regulates the aggressive property including migration and cell matrix attachment of breast cancer with the involvement of HSP90A. Citation Format: Zhe Zhao, Tracey A. Martin, Johannes Benedikt, Shuo Cai, Lin Ye, Eleri Davies, Robert E Mansel, Wen G. Jiang. Kinase D interacting substrate 220 (Kidins220) and disease progression of breast cancer, the role of heat shock protein90 (HSP90) [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P3-01-18.

Mucosal delivery of live Lactococcus lactis expressing functionally active JlpA antigen induces potent local immune response and prevent enteric colonization of Campylobacter jejuni in chickens

Successful colonization of the mucosal epithelial cells is the key early step for Campylobacter jejuni (C. jejuni) pathogenesis in humans. A set of Surface Exposed Colonization Proteins (SECPs) are known to take leading role in bacterial adhesion and subsequent host pathogenesis. Among the major SECPs, the constitutively expressed C. jejuni surface lipoprotein Jejuni lipoprotein A (JlpA), interacts with intestinal heat shock protein 90α (Hsp90α) and contributes in disease progression by triggering pro-inflammatory responses via activation of NF-κB and p38 MAP kinase pathways. In addition to its ability to express on the surface, high sequence conservation of JlpA protein among different Campylobacter spp make it a suitable vaccine target against C. jejuni. Given that chickens are the primary source for C. jejuni infection in humans and persistent cecal colonization significantly contribute in pathogen transmission, we explicitly used chickens as a model to test the immune-protective efficacy of JlpA protein. Taking into account that gastro-intestinal tract is the major site for C. jejuni colonization, we chose to use mucosal (intragastric) route as mode for JlpA antigen delivery. To deliver JlpA via mucosal route, we engineered a food grade Lactic acid producing bacteria, Lactococcus lactis (L. lactis) to express functionally active JlpA protein in the surface. Further, we demonstrated its ability to substantially improve the antigen specific local immune responses in the intestine along with significant immune-protection against enteric colonization of C. jejuni in chickens.

Metformin Inhibits Tumor Metastasis through Suppressing Hsp90α Secretion in an AMPKα1-PKCγ Dependent Manner.

Metformin has been documented in epidemiological studies to mitigate tumor progression. Previous reports show that metformin inhibits tumor migration in several cell lines, such as MCF-7 and H1299, but the mechanisms whereby metformin exerts its inhibitory effects on tumor metastasis remain largely unknown. The secreted proteins in cancer cell-derived secretome have been reported to play important roles in tumor metastasis, but whether metformin has an effect on tumor secretome remains unclear. Here we show that metformin inhibits tumor metastasis by suppressing Hsp90α (heat shock protein 90α) secretion. Mass spectrometry (MS) analysis and functional validation identify that eHsp90α (extracellular Hsp90α) is one of the most important secreted proteins for metformin to inhibit tumor cells migration, invasion and metastasis both in vitro and in vivo. Moreover, we find that metformin inhibits Hsp90α secretion in an AMPKα1 dependent manner. Our data elucidate that AMPKα1 (AMP-activated protein kinase α1) decreases the phosphorylation level of Hsp90α by inhibiting the kinase activity of PKCγ (protein kinase Cγ), which suppresses the membrane translocation and secretion of Hsp90α. Collectively, our results illuminate that metformin inhibits tumor metastasis by suppressing Hsp90α secretion in an AMPKα1 dependent manner.