Search

Glioblastoma multiforme (GBM) is the most common primary brain tumor, characterized by a remarkable inner complexity and inter-tumor variability. Moreover, it is very aggressive and resistant to conventional treatments, so that it rapidly relapse. Therefore, there is an immediate need for experimental strategies to enhance our comprehension of GBM, aiming to mitigate its economic and social impact. Here, we described different in vivo and in vitro strategies currently used for the study of GBM. First, we gave a brief and general overview of the classical in vivo models, including xenograft mouse and zebrafish models and canine models, offering a wide range of advantages but also presenting a series of strong limitations. Thus, we described in vitro models, starting from more traditional 2D culture models, comparing different approaches, and critically exposing the advantages and disadvantages of using one or the other methods. We also briefly described GBM 2D culture systems that allow recreating multiple cell-cell and cell-extracellular matrix contacts but still do not reflect the complexity of in vivo tumors. We finally described the intricacies of the more novel 3D in vitro models, e.g., spheroids and organoids. These sophisticated models have demonstrated exceptional suitability across a wide spectrum of applications in cancer research, ranging from fundamental scientific inquiries to applications in translational research. Their adaptability and three-dimensional architecture render them invaluable tools, offering new insights and paving the way for advancements in both basic and applied research. [ABSTRACT FROM AUTHOR]

The fundamental challenge in fighting cancer is the development of protective agents able to interfere with the classical pathways of malignant transformation, such as extracellular matrix remodeling, epithelial–mesenchymal transition and, alteration of protein homeostasis. In the tumors of the brain, proteotoxic stress represents one of the main triggering agents for cell transformation. Curcumin is a natural compound with anti-inflammatory and anti-cancer properties with promising potential for the development of therapeutic drugs for the treatment of cancer as well as neurodegenerative diseases. Among the mediators of cancer development, HSP60 is a key factor for the maintenance of protein homeostasis and cell survival. High HSP60 levels were correlated, in particular, with cancer development and progression, and for this reason, we investigated the ability of curcumin to affect HSP60 expression, localization, and post-translational modifications using a neuroblastoma cell line. We have also looked at the ability of curcumin to interfere with the HSP60/HSP10 folding machinery. The cells were treated with 6, 12.5, and 25 µM of curcumin for 24 h, and the flow cytometry analysis showed that the compound induced apoptosis in a dose-dependent manner with a higher percentage of apoptotic cells at 25 µM. This dose of curcumin-induced a decrease in HSP60 protein levels and an upregulation of HSP60 mRNA expression. Moreover, 25 µM of curcumin reduced HSP60 ubiquitination and nitration, and the chaperonin levels were higher in the culture media compared with the untreated cells. Furthermore, curcumin at the same dose was able to favor HSP60 folding activity. The reduction of HSP60 levels, together with the increase in its folding activity and the secretion in the media led to the supposition that curcumin might interfere with cancer progression with a protective mechanism involving the chaperonin. [ABSTRACT FROM AUTHOR]

With this invited commentary we want to draw the attention of young medical doctors, the main readers of this journal, towards the existence and importance of a group of nanovesicles released by human cells: the exosomes. These vesicles are incontinently secreted as a mean of cell-to-cell communication. They are involved in a number of physiologic processes as well as in the pathogenesis of, virtually, all human diseases. They can be isolated from all biological fluids, like blood, urine, sweat, sperm, crevicular fluid, bile, etc., and their composition in terms of proteins, RNA and lipids is different in pathology that in physiologic conditions. It is therefore possible to predict that they will become an important diagnostic and therapeutic tool in medicine. [ABSTRACT FROM AUTHOR]

The article offers information about the role of extracellular heat shock proteins (Hsps) in cancer theranostics. It mentions that how extracellular Hsps act as early cancer biomarkers, along with discusses that extracellular Hsps can be considered as targets for modern and effective anticancer therapies.

Papillary thyroid carcinoma (PTC) is the most frequently occurring subtype of thyroid cancer. Exosomes (EXs) secreted from cells to the extracellular environment play an important role in intercellular communication in normality and pathology. Recent data indicates that tumor cells-derived EXs contribute to cancer progression through the modulation of tumor microenvironment [1]. Heat Shock Protein (HSPs) are often overexpressed during carcinogenesis and different studies shown that they can be released by tumors cells and that the mechanism of release is mediated by EXs pathway. In this project we performed an immunomorphological study to investigate Hsp60, 90,70,27 levels expression profile in thyroid tissue from patients with benign goiter (used as benign desease) and patients with PTC. Moreover for each patient, blood samples were collected before and a one week after surgery, to obtain EXs. We performed Western Blotting analysis to verify the presence and the levels of the same HSPs. The immunoistochemistry shown an overexpression of Hsp60,90 and 27 in the PTC cases comparison with peritumoral tissue and with goiter cases. Instead the Hsp70 levels showed no significant changes. In particular Hsp60, 90 and 27 were visible at cytoplasmic and membrane levels. Data regarding exosomal fraction assessment by standard methods (TEM, and WB analysis for Alix) to identify exosomes confirmed their identity. The levels of Hsp60, 90,27 in the exosomes of patients with PTC before surgery were significantly higher than in the exosomes from the same patients after surgery. The data obtained shown that, as demonstrated in other cancer type [2], the HSP levels studied increased in PTC specimens respect to goiter specimens. Moreover the membrane localization of these HSP suggested a their release in tumor microenviroment, in fact we observed exosomal HSP before surgery in PTC patients. The HSP decreases after surgery indicated that if disease recurrence occurs, HSP levels will increase again. For this reason we hipotized that chaperonins could be good candidates as biomarkers of PTC. [ABSTRACT FROM AUTHOR]

Among the mediators of stress response, Heat Shock Proteins (HSPs) play essential roles in cell survival, protein folding, trafficking and degradation [1]. In particular, HSPs alterations were associated with temporal lobe epilepsy (TLE) [2] and recently, specific microRNAs (miRNA) have been proposed as regulators of HSPs expression [3]. The significance of HSP60 in hippocampus, derived from patients affected by drug resistant TLE with hippocampal sclerosis and associated controls, was investigated by immunohistochemistry while circulating levels of this protein were detected by ELISA test. qRT-PCR was used to evaluate the expression levels of HSP60 and associated miRNA such as miR1 and miR206 in hippocampus. Moreover, miR-8071, miR-663, miR-146a and miR-124 expression levels associated with clinical features of TLE were also investigated. Our findings show that HSP60 is localized inside neurons somata and neuropil. Hsp60 expression levels were correlated to those of miR1 and miR206. Moreover, plasma Hsp60 levels in patients were higher than those of controls. Finally, circulating levels of miR-8071, miR-663, miR-146a and miR-124 decreased in TLE patients and were correlated to neuroinflammation and seizure recurrences. Our work suggests that Hsp60 and associated miRNA levels are altered in relation to epileptogenesis and disease progression and may serve as a target for new therapeutic approaches in the management of TLE patients. [ABSTRACT FROM AUTHOR]

One of the many pathologic conditions still without a satisfactory solution is that of brain tumors. The prognosis is poor even after surgical resection followed by post-operatory chemoand radio-therapie [1]. It is, therefore, cogent to find innovative treatment tools. Three recent developments may provide elements to discover novel treatment strategies and means. These developments are: the discovery that molecular chaperones can be determinant factors in the process of tumorigenesis [2]; the elucidation of the role of miRNAs in gene regulation and determination of protein functions, including molecular chaperones; in the various cell compartments [3]; the increasing understanding and characterization of exosomes (exo), particularly in what refers to their release by tumor cells, contents including chaperones and miRNA, and ability to travel and interact with target cells near their origin or far [4]. The aim of the current study is to research a particular molecular chaperone, the HSP60 presence, levels, expression and distribution in tumor and peritumoral cells of primary brain tumors in vivo. The presence and level of HSP60 and some miRNAs involved in his regulation in exo isolated by blood samples obtained from patients with cancer before and after ablative surgery were also investigated. A total of 45 brain surgeries were performed. Blood and pathological tissue sample were taken from patient on the day of the surgery. For each patient, blood samples were collected at one week, one month and three months after surgery. Blood samples were collected from each patients and processed for plasma isolation, from which exo were isolated. The tumor and normal tissue section were used to perform the immunomorphological analyses and was assessed the valuation of HSP60 and microRNAs HSP60-related in exo obtained from blood of patients. Our work provided evidences about presence and levels of the main miRNA involved in HSP60 regulation in tumor brain, which would be useful in detecting the disease and monitoring its progression. [ABSTRACT FROM AUTHOR]

Heat-shock protein (Hsp)60 is a mitochondrial protein involved in assisting the correct folding of other mitochondrial client proteins [1]. Recently, this chaperonine has been considered as an emerging target for Alzheimer's Disease (AD) because seems to be able to mediate the translocation of Amyloid Precursor Protein (APP) and Amyloid Beta peptide (Aß) to the mitochondria [2]. The fundamental challenge on fighting the Alzheimer's Disease (AD) is the development of neuro-protective agents, able to interfere with biochemical pathways responsible for the protein aggregation process whose clinical signature is represented by the plaques deposition. In this study we investigated the effect of curcumin, an emerging lead-compound for the development of neuro-protective drugs, on Hsp60 gene, protein expression and folding activity using a neuroblastoma cell line (LAN5). We demonstrated that the treatment of LAN5 cells with curcumin caused a down-regulation of mitochondrial Hsp60 protein and gene expression. On the other hand, curcumin enhanced the folding activity of the chaperonine. The ability of curcumin to affect Hsp60 expression as well as its ability to interact with the Hsp60/Hsp10 folding machine, open new frontiers in the use of putative therapeutic properties of curcumin as a switch from cancer therapy to AD treatment. [ABSTRACT FROM AUTHOR]

Heat shock proteins (Hsps) are highly expressed in a variety of cancer types contributing to tumor cell propagation and protection against apoptosis [1]. The current anti-cancer therapy is not always target specific and often is associated with complications for patients, Therefore new effective, specific and less toxic therapeutic approaches are needed. Medicinal mushrooms have emerged as wonderful source of nutraceuticals, anti-oxidants, anticancer, prebiotic, anti-inflammatory, cardiovascular, anti-microbial, and anti-diabetic. The ongoing research projects are aimed to promote mushrooms as new generation ''biotherapeutics'' [2]. The aim of this study was to evaluate whether the cold-water extracts of Pleurotus eryngii var. eryngii can affect Hsp90, 70, 60 and 27 levels in an in vitro model of colon cancer (C26 cells). Cell viability was evaluated using MTT assay after treating the cells with different concentrations of extracts (0-1.9 µg/µl) in the culture medium for 24 and 48 hours. Hsp90, 70, 60 and 27 levels were measured using western blotting and immunofluorescence analysis. Moreover, we evaluated the anticancer effect of the P. eryngii var. eryngii extract in an animal model of ectopicallyimplanted C26 colon carcinoma, widely used as an experimental model of cancer cachexia. We prepared a mixture of lyophilized P. eryngii var. eryngii with the mice standard diet and the animals were daily fed with ~4g of the mix until they died to draw a survival curve. We sampled the neoformations grown after implantation e on these we performed an immunohistochemistry for Hsp60. Our results showed that the extract significantly decreased cells viability at 0.48 µg/µl after both 24 and 48 hours of treatments. Western blotting analysis and immunofluorescence showed that Hsp60 protein levels were down-regulate at 24h of treatment but increased after 48h. On the contrary, Hsp90, 70 and 27 protein levels did not changed. In the in vivo model, P. eryngii var. eryngii in the diet significantly extended the median survival compared to untreated mice. The immunoistochemical experiments suggested that Pleuery significantly affected the increase of Hsp60 protein levels. These preliminary results are promising for further studies to better understand the potential effects of P. eryngii var. eryngii on cancer progression especially regarding Hsp60 role. [ABSTRACT FROM AUTHOR]

Fatty acid-binding proteins (FABPs) are lipid chaperones assisting in the trafficking of long-chain fatty acids with functions in various cell compartments, including oxidation, signaling, gene-transcription regulation, and storage. The various known FABP isoforms display distinctive tissue distribution, but some are active in more than one tissue. Quantitative and/or qualitative changes of FABPs are associated with pathological conditions. Increased circulating levels of FABPs are biomarkers of disorders such as obesity, insulin resistance, cardiovascular disease, and cancer. Deregulated expression and malfunction of FABPs can result from genetic alterations or posttranslational modifications and can be pathogenic. We have assembled the disorders with abnormal FABPs as chaperonopathies in a distinct nosological entity. This entity is similar but separate from that encompassing the chaperonopathies pertaining to protein chaperones. In this review, we discuss the role of FABPs in the pathogenesis of metabolic syndrome, cancer, and neurological diseases. We highlight the opportunities for improving diagnosis and treatment that open by encompassing all these pathological conditions within of a coherent nosological group, focusing on abnormal lipid chaperones as biomarkers of disease and etiological-pathogenic factors.

Background: Alzheimer's disease (AD) is a dementia, a neurodegenerative condition, and a protein-misfolding disease or proteinopathy, characterized by protein deposits, extracellular plaques and intracellular neurofibrillary tangles, which contain the AD's typical pathological proteins, abnormal β-amyloid and hyperphosphorylated tau, respectively, and are located predominantly in the cortex of the frontal, parietal, and temporal brain lobes. What is the role of molecular chaperones in AD? Data indicate that molecular chaperones, also known as Hsp, are involved in AD, probably displaying protective roles and/or acting as pathogenic factors as it occurs in chaperonopathies in which case AD would be suitable to chaperonotherapy. Hsp60, Hsp70, and Hsp90 can be augmented and overexpressed or diminished and downregulated in various situations in AD affected tissues and cells, indicating they are active during disease development and progression., Question: What is the role of molecular chaperones in AD? Data indicate that molecular chaperones, also known as Hsp, are involved in AD, probably displaying protective roles and/or acting as pathogenic factors as it occurs in chaperonopathies in which case AD would be suitable to chaperonotherapy., Objective: Investigate the role of Hsp in AD, focusing on Hsp60, Hsp70, and Hsp90., Method: Critical examination of published data., Results: Hsp60, Hsp70, and Hsp90 can be augmented and overexpressed or diminished and downregulated in various situations in AD affected tissues and cells, indicating they are active during disease development and progression., Conclusion and Perspectives: Notwithstanding that the roles and mechanisms of action of chaperones in AD are still incompletely understood, there is already enough evidence to encourage the development of therapeutic strategies targeting them, either to block their activity in case they promote disease progression or to boost their performance when they are protective. The latter is an example of positive chaperonotherapy, which also includes chaperone replacement via gene or protein administration. On the contrary, if a chaperone is found to help the disease, it has to be blocked or eliminated, which constitute modalities of negative chaperonotherapy.

Background: Heat shock proteins (Hsps) assist other proteins in their folding and drive the degradation of defective proteins. During evolution, these proteins have also acquired other roles. Hsp10 is involved in immunomodulation and tumor progression. Hsp90 stabilizes a range of "client" proteins involved in cell signaling. The present study evaluated the expression levels of Hsp10 and Hsp90 in normal mucosa and adenocarcinoma samples of human large bowel., Materials and Methods: Samples of normal mucosa and adenocarcinoma were collected and Reverse transcriptase-polymerase chain reaction RT-PCR, western blotting (WB) analyses, as well as immunohistochemistry were performed to evaluate the expression levels of Hsp10 and Hsp90., Results: RT-PCR showed a higher gene expression of Hsp10 and Hsp90 in adenocarcinoma samples compared to healthy mucosa. WB results confirmed these findings. Immunohistochemistry revealed higher levels of Hsp10 in adenocarcinoma in both the epithelium and the lamina propria, while Hsp90 expression was higher in the adenocarcinoma samples only in the lamina propria., Conclusion: Hsp10 and Hsp90 may be involved in large bowel carcinogenesis., (Copyright© 2014 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.)