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Exosome characterization. (A) Quantification and characterization by Western blotting (WB) of markers of exosomes from patients with goiter and papillary carcinoma before surgery (BS) and after surgery (AS). (B) Dynamic light scattering (DLS) characterization of exosomes showing their concentrations and their diameters. Mean ± SE of three different experiments are shown. (C) Transmission electron microscopy (TEM) (bar 200 nm) and (D) atomic force microscopy (AFM) images showing the typical characteristics of exosomes.
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The thyroid is a major component of the endocrine system and its pathology can cause serious diseases, e.g., papillary carcinoma (PC). However, the carcinogenic mechanisms are poorly understood and clinical useful biomarkers are scarce. Therefore, we determined if there are quantitative patterns of molecular chaperones in the tumor tissue and circu...
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... were purified from plasma from BG patients, and from plasma from PC patients before and after ablative surgery. The levels of the exosome markers Alix and CD81 were assessed in all exosomes by Western blotting (WB) ( Figure 3A). Dynamic light scattering (DLS) quantitative analysis showed that the mean value of the number of plasmatic exosomes of patients with BG was 7.213 × 10 13 ± 2.394 × 10 13 and the mean value of sizes was 41.012 ± 7.739 nm. ...
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... PC patients, the number of plasmatic exosomes after surgery decreased from 3.294 × 10 13 ± 1.339 × 10 13 to 2.223 × 10 13 ± 8.163 × 10 12 , but the difference, although appreciable, was not statistically significant. On the contrary, in PC the size of exosomes after surgery was significantly greater than that before it ( Figure 3B). Transmission electron microscopy (TEM) and atomic force microscopy (AFM) images demonstrated the typical exosomal features in all our preparations ( Figure 3C,D). ...
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... the contrary, in PC the size of exosomes after surgery was significantly greater than that before it ( Figure 3B). Transmission electron microscopy (TEM) and atomic force microscopy (AFM) images demonstrated the typical exosomal features in all our preparations ( Figure 3C,D). ...
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... were purified from plasma from BG patients, and from plasma from PC patients before and after ablative surgery. The levels of the exosome markers Alix and CD81 were assessed in all exosomes by Western blotting (WB) ( Figure 3A). Dynamic light scattering (DLS) quantitative analysis showed that the mean value of the number of plasmatic exosomes of patients with BG was 7.213 × 10 13 ± 2.394 × 10 13 and the mean value of sizes was 41.012 ± 7.739 nm. ...
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... PC patients, the number of plasmatic exosomes after surgery decreased from 3.294 × 10 13 ± 1.339 × 10 13 to 2.223 × 10 13 ± 8.163 × 10 12 , but the difference, although appreciable, was not statistically significant. On the contrary, in PC the size of exosomes after surgery was significantly greater than that before it ( Figure 3B). Transmission electron microscopy (TEM) and atomic force microscopy (AFM) images demonstrated the typical exosomal features in all our preparations ( Figure 3C,D). ...
Citations
... Following their accumulation within the cell, HSPs can be actively released into the extracellular space and into the bloodstream. The extracellular HSP60 levels in exosomes were significantly higher compared to those in exosomes from post-surgery papillary carcinoma patient's and patients with benign goiter (BG), suggesting their potential as biomarkers for clinical applications [152]. In colon cancer, HSP60 is localized in the pericellular interstitium of affected tissue on macrophages and NK cells and is released into circulation through HSP-EVs. ...
Molecular chaperones are highly conserved across evolution and play a crucial role in preserving protein homeostasis. The 60 kDa heat shock protein (HSP60), also referred to as chaperonin 60 (Cpn60), resides within mitochondria and is involved in maintaining the organelle’s proteome integrity and homeostasis. The HSP60 family, encompassing Cpn60, plays diverse roles in cellular processes, including protein folding, cell signaling, and managing high-temperature stress. In prokaryotes, HSP60 is well understood as a GroEL/GroES complex, which forms a double-ring cavity and aids in protein folding. In eukaryotes, HSP60 is implicated in numerous biological functions, like facilitating the folding of native proteins and influencing disease and development processes. Notably, research highlights its critical involvement in sustaining oxidative stress and preserving mitochondrial integrity. HSP60 perturbation results in the loss of the mitochondria integrity and activates apoptosis. Currently, numerous clinical investigations are in progress to explore targeting HSP60 both in vivo and in vitro across various disease models. These studies aim to enhance our comprehension of disease mechanisms and potentially harness HSP60 as a therapeutic target for various conditions, including cancer, inflammatory disorders, and neurodegenerative diseases. This review delves into the diverse functions of HSP60 in regulating proteo-homeostasis, oxidative stress, ROS, apoptosis, and its implications in diseases like cancer and neurodegeneration.
... Among the proteins present in the cargo of EVs, proteins belonging to the Chaperoning System (CS), with most of them being HSPs, are found [87]. These proteins are necessary for the maintenance of cellular homeostasis and they are also involved in several stages of carcinogenesis [48,[100][101][102][103]. In glioma, various heat shock proteins (Hsps) play significant roles in promoting malignant behavior. ...
Glioblastoma multiforme (GBM) stands out as the most tremendous brain tumor, constituting 60% of primary brain cancers, accompanied by dismal survival rates. Despite advancements in research, therapeutic options remain limited to chemotherapy and surgery. GBM molecular heterogeneity, the intricate interaction with the tumor microenvironment (TME), and non-selective treatments contribute to the neoplastic relapse. Diagnostic challenges arise from GBM advanced-stage detection, necessitating the exploration of novel biomarkers for early diagnosis. Using data from the literature and a bioinformatic tool, the current manuscript delineates the molecular interplay between human GBM, astrocytes, and myeloid cells, underscoring selected protein pathways belonging to astroglia and myeloid lineage, which can be considered for targeted therapies. Moreover, the pivotal role of extracellular vesicles (EVs) in orchestrating a favorable microenvironment for cancer progression is highlighted, suggesting their utility in identifying biomarkers for GBM early diagnosis.
... CircRNAs are a class of ncRNAs with a covalently closed circular form and without a 5′ tail or a 3′ head. Whole genome sequencing has shown that circRNAs are broadly expressed in various tissues and cell lines (27,28) and are involved in a wide range of biological processes, such as those involved in tumors. ...
The use of liquid biopsy in cancer research has grown exponentially, offering potential for early detection, treatment stratification, and monitoring residual disease and recurrence. Exosomes, released by cancer cells, contain tumor-derived materials and are stable in biofluids, making them valuable biomarkers for clinical evaluation. Bibliometric research on osteosarcoma (OS) and exosome-derived diagnostic biomarkers is scarce. Therefore, we aimed to conduct a bibliometric evaluation of studies on OS and exosome-derived biomarkers. Using the Web of Science Core Collection database, Microsoft Excel, the R “Bibliometrix” package, CiteSpace, and VOSviewer software, quantitative analyses of the country, author, annual publications, journals, institutions, and keywords of studies on exosome-derived biomarkers for OS from 1995 to 2023 were performed. High-quality records (average citation rate ≥ 10/year) were filtered. The corresponding authors were mainly from China, the USA, Australia, and Canada. The University of Kansas Medical Center, National Cancer Center, Japan, and University of Kansas were major institutions, with limited cooperation reported by the University of Kansas Medical Center. Keyword analysis revealed a shift from cancer progression to mesenchymal stem cells, exosome expression, biogenesis, and prognostic biomarkers. Qualitative analysis highlighted exosome cargo, including miRNAs, circRNAs, lncRNAs, and proteins, as potential diagnostic OS biomarkers. This research emphasizes the rapid enhancement of exosomes as a diagnostic frontier, offering guidance for the clinical application of exosome-based liquid biopsy in OS, contributing to the evolving landscape of cancer diagnosis.
... High levels of EVs in circulation are dependent on tumor presence because, after tumor removal, HSP60-EVSs in circulation decrease [83]. The same scenario is found in other tumor types; in thyroid papillary carcinomas, HSP27-, HSP60-, and HSP90-EV levels in plasma decreased in number after surgical resection of the tumor [84]. These data support the idea that HSP60-EVs in circulation may be useful to follow up in patients' recurrence after surgical treatments, for instance. ...
From an evolutive perspective, tumor cells endure successive turnover upon stress conditions and pressure to adapt to new environments. These cells use exceptional communication skills to share biological information to “survive upon every metabolic cost”. The tumor microenvironment (TME) is a miscellaneous collection of cells, factors, and extracellular vesicles (EVs). EVs are small lipid bilayer-delimited particles derived from cells with sizes ranging from 100 to 1000 nm. Exosomes (<160 nm) are the minor subtype of EVs, originating from the endosomal pathways. The TME also contains “giant” vesicles, microvesicles (100–1000 nm, MV), originated from membrane blebbing. EVs can act as intercellular communication mediators, contributing to many biological processes, by carrying different biomolecules, such as proteins, lipids, nucleic acids, and metabolites. EV secretion can promote either tumor cell survival or manage their stress to death. Tumor-derived EVs transfer adaptative stress signaling to recipient cells, reprograming these cells. Heat shock proteins (HSP) are prominent stress response regulators, specifically carried by exosomes. HSP-loaded EVs reprogram tumor and TME cells to acquire mechanisms contributing to tumor progression and therapy resistance. The intercellular communication mediated by HSP-loaded EVs favors the escape of tumor cells from the endoplasmic reticulum stress, hypoxia, apoptosis, and anticancer therapies. Extracellular HSPs activate and deactivate the immune response, induce cell differentiation, change vascular homeostasis, and help to augment the pre-metastatic niche formation. Here we explore EVs’ mechanisms of HSP transmission among TME cells and the relevance of these intercellular communications in resistance to therapy.
... Hsp90 is the master regulator of various growth, inflammation, and survival pathways [8]. Pronounced expression of Hsp90 in several cancer types has been reported [26][27][28][29][30]. Hsp90 has been proposed as a potential biomarker because its increased expression in neoplastic tissue correlates with clinically advanced stages and poor prognosis [31][32][33][34][35][36]. ...
The chaperone system (CS) is emerging as a key multistage participant in carcinogenesis. The CS chief components are the molecular chaperones (some of which are named heat shock proteins or Hsp), which are typically cytoprotective but if abnormal in structure, location, or quantity, can become etiopathogenic and cause diseases, known as chaperonopathies, including some cancers. For example, abnormal Hsp90 expression is associated with tumorigenesis and poor prognosis. Hsp90 is positioned at the center of several key oncogenic pathways by stabilizing and activating oncogenic kinases responsible for driving cell proliferation and survival. Consequently, inhibition of Hsp90 is being investigated as a possible anti-cancer strategy and some results are encouraging. However, the 5-year survival rate for patients suffering from salivary gland carcinomas is still unsatisfactory. Because of the rarity of these malignancies, they may have been overlooked and understudied and, thus, novel therapies (e.g., inhibition of CS components like Hsp90 and others) are urgently needed. In this review, we also summarize the histopathological quantitative patterns and the intra- and extra-cellular location characteristics of Hsp90 in tumors of salivary glands, pointing to their potential for differential diagnosis, prognostication, and patient monitoring.
... They revealed that in PTC patients with LNM serum exosomes, integrin-associated proteins, such as SRC, TLN1, ITGB2, ACTB, and CAPNS1, were clearly upregulated, which suggested that PTC is associated with distinct changes in protein signaling. In addition, another study [134] assessed the level of plasma exosomes in PTC and benign goiter patients before and after ablative surgery. They revealed that PTC had higher levels of exosomal Hsp90, Hsp60, and Hsp27 than benign goiter, and they were also increased in PTC before and after ablative surgery, suggesting their potential as biomarkers for clinical applications. ...
Thyroid cancer has become more common in recent years all around the world. Many issues still need to be urgently addressed in the diagnosis, treatment, and prognosis of thyroid cancer. Liquid biopsy (mainly circulating tumor DNA (ctDNA), circulating tumor cells (CTCs), and circulating exosomes) may provide a novel and ideal approach to solve these issues, allows us to assess the features of diseases more comprehensively, and has a function in a variety of malignancies. Recently, liquid biopsy has been shown to be critical in thyroid cancer diagnosis, treatment, and prognosis in numerous previous studies. In this review, by testing CTCs, ctDNA, and exosomes, we focus on the possible clinical role of liquid biopsy in thyroid cancer, including diagnostic and prognostic biomarkers and response to therapy. We briefly review how liquid biopsy components have progressed in thyroid cancer by consulting the existing public information. We also discuss the clinical potential of liquid biopsy in thyroid cancer and provide a reference for liquid biopsy research. Liquid biopsy has the potential to be a useful tool in the early detection, monitoring, or prediction of response to therapies and prognosis in thyroid cancer, with promising clinical applications.
... In other words, it prevents the abnormal misfolding or aggregation of proteins [7], aids in the process of protein migration to intracellular organelles, and breaks down denatured proteins [8]. HSP60 plays a role in tumor growth and progression in several cancers, including ovarian [9], gastric cancer [10], liver cancer [11], pancreas cancer [12], breast cancer [13], papillary thyroid carcinoma [14], cervical cancer [15], head and neck cancer [16], and CRC [17][18][19]. However, there are conflicting reports that increased HSP60 expression correlates with better oncological outcomes [20,21]. ...
Simple Summary
HSP60, a mitochondrial chaperone, can promote or inhibit cancer progression. Patients with colorectal cancer (CRC) were examined for HSP60 expression using the TNM classification. Patients with differentiated and p53-mutated CRC expressed high levels of HSP60. Compared with patients with high HSP60 expression, those with low expression had event-free and disease-specific survival hazard ratios of 1.42 and 1.69, respectively. TNM class and HSP60 expression affected survival, especially in late/advanced stages. The expression of the HSPD1 gene, which encodes HSP60, exhibited the same pattern as the protein. The hazard ratios for overall and relapse-free survival were 1.80 and 1.87, respectively, for patients with reduced HSPD1 expression. Low HSPD1 expression and advanced malignancy worsen CRC prognosis. This study suggests that HSPD1/HSP60 may be a useful biomarker for refined survival prediction in late-stage and advanced-stage CRC, allowing for individualized therapy.
Abstract
The role of heat shock protein 60 (HSP60), a mitochondrial chaperone, in tumor progression or its anti-tumor effects remains controversial. This study aimed to confirm the possibility of using HSP60 as a prognostic marker in patients with colorectal cancer (CRC), considering TNM classification for precise prediction. HSP60 expression increased with differentiation and p53 mutations in patients. However, compared to patients with high HSP60 expression, patients with low HSP60 expression had event-free survival and disease-specific survival hazard ratios (HRs) of 1.42 and 1.69, respectively. Moreover, when the survival rate was analyzed by combining TNM classification and HSP60 expression, the prognosis was poor, particularly when HSP60 expression was low in the late/advanced stage. This pattern was also observed with HSP family D member 1, HSPD1, the gene that encodes HSP60. Low HSPD1 expression was linked to lower overall survival and relapse-free survival rates, with HRs of 1.80 and 1.87, respectively. When TNM classification and HSPD1 expression were considered, CRC patients with low HSPD1 expression and advanced malignancy had a poorer prognosis than those with high HSPD1 expression. Thus, HSPD1/HSP60 can be a useful biomarker for a sophisticated survival prediction in late- and advanced-stage CRC, allowing the design of individualized treatment strategies.
... It has been shown that the profile of HSPs in papillary thyroid cancer tissue and circulating exosomes are identical; in particular, HSP27 and HSP60 predominate, while HSP90 and HSP70 are weakly expressed [27]. Although Western blotting was used to determine the level of HSPs in exosomes in [27] and flow cytometry was used in our work, we obtained similar data on the distribution of HSPs in sEVs of patients with pretumor disease and CRC, indicating the predominance of HSP60 and HSP27 on of both CD9-positive sEVs and FABP4-positive sEVs. ...
... It has been shown that the profile of HSPs in papillary thyroid cancer tissue and circulating exosomes are identical; in particular, HSP27 and HSP60 predominate, while HSP90 and HSP70 are weakly expressed [27]. Although Western blotting was used to determine the level of HSPs in exosomes in [27] and flow cytometry was used in our work, we obtained similar data on the distribution of HSPs in sEVs of patients with pretumor disease and CRC, indicating the predominance of HSP60 and HSP27 on of both CD9-positive sEVs and FABP4-positive sEVs. ...
The majority of colorectal cancer patients (CRCPs) develop tumors on the background of “metabolically healthy obesity” or metabolic syndrome. The aim of the work was to study the levels of matrix metalloproteinases (MMPs) and heat shock proteins (HSPs) on the surface of blood plasma CD9-positive and FABP4-positive small extracellular vesicles (sEVs) from CRCPs depending on metabolic status and tumor angiogenesis, as well as to evaluate the sEVs markers as predictors of the effectiveness of thermoradiotherapy. In CRCPs, compared with patients with colorectal polyps (CPPs), the proportion of triple positive EVs and EVs with the MMP9+MMP2-TIMP1+ phenotype increased significantly among FABP4-positive EVs (adipocyte-derived EVs), which in general may indicate the overexpression of MMP9 and TIMP1 by adipocytes or adipose tissue macrophages in CRCPs. The results obtained have prospects for use as markers to clarify cancer risk in CPPs. One can assume that for CRCPs with metabolic syndrome or metabolically healthy obesity, it is the FABP4+MMP9+MMP2-TIMP1- population of circulating sEVs that is the most optimal biomarker reflecting tumor angiogenesis. Determining this population in the blood will be useful in monitoring patients after treatment for the early detection of tumor progression. CD9+MMP9+MMP2-TIMP1- and MMP9+MMP2-TIMP1+ subpopulations of circulating sEVs are the most promising predictors of the efficacy of thermoradiation therapy because their levels at baseline differ significantly in CRCPs with different tumor responses.
... This becomes a vicious cycle where an increase in mutated proteins drives the induction of Hsps, increase in translational processes and expression of proteins leading to transformation (Ciocca et al. 2013). Pronounced expression of Hsp90 in several cancer types has been reported (Rappa et al. 2014;Kamm et al. 2019;Caruso Bavisotto et al. 2019;Gorska-Ponikowska et al. 2020;Barone et al. 2021). Therefore, Hsp90 has been proposed as a potential candidate for biomarker studies as its increased expression in neoplastic tissue correlated directly with clinically advanced stages and poor prognosis (Pick et al. 2007;Chiu et al. 2011;Chen et al. 2015;A. ...
The chaperone system (CS) of an organism is composed of molecular chaperones, chaperone co-factors, co-chaperones, and chaperone receptors and interactors. It is present throughout the body but with distinctive features for each cell and tissue type. Previous studies pertaining to the CS of the salivary glands have determined the quantitative and distribution patterns for several members, the chaperones, in normal and diseased glands, focusing on tumors. Chaperones are cytoprotective, but can also be etiopathogenic agents causing diseases, the chaperonopathies. Some chaperones such as Hsp90 potentiate tumor growth, proliferation, and metastasization. Quantitative data available on this chaperone in salivary gland tissue with inflammation, and benign and malignant tumors suggest that assessing tissue Hsp90 levels and distribution patterns is useful for differential diagnosis-prognostication, and patient follow up. This, in turn, will reveal clues for developing specific treatment centered on the chaperone, for instance by inhibiting its pro-carcinogenic functions (negative chaperonotherapy). Here, we review data on the carcinogenic mechanisms of Hsp90 and their inhibitors. Hsp90 is the master regulator of the PI3K-Akt-NF-kB axis that promotes tumor cell proliferation and metastasization. We discuss pathways and interactions involving these molecular complexes in tumorigenesis and review Hsp90 inhibitors that have been tested in search of an efficacious anti-cancer agent. This targeted therapy deserves extensive investigation in view of its theoretical potential and some positive practical results and considering the need of novel treatments for tumors of the salivary glands as well as other tissues.
... Therefore, investigation of EVs is useful to highlight mechanisms that may determine health or disease. Furthermore, as EVs carry genetic material and proteins that may affect different signaling pathways in the target cells and organs, and since they are commonly found in biological fluids and tissue, they can be used as noninvasive and easily accessible disease biomarkers [48][49][50][51][52][53]. As stated above, in different diseases, cells can produce EVs with alterations in number and molecular content compared to physiological conditions, and accumulating evidence demonstrates their role in cancer, in which the application of EVs may be helpful for early diagnostics and the identification of new therapeutic targets [52]. ...
... Among proteins transported within the EVs, there are metalloproteinases, growth factors and chemokines, used as secondary messengers for the coordination of cellular responses [123]. Several studies have shown that the release of EVs could play a role in response to vaccination and therapeutic applications, as a vehicle in the delivery of drugs and targeted therapy [50,[124][125][126]. In general, IEC-derived EVs are able to regulate the integrity of the epithelial barrier thanks to the transport of desmosomal cadherins that stabilize cell-cell epithelial adhesions, as well as being able to protect against pathogenic infections thanks to the transport of some antimicrobial peptides, such as beta-defensin [127,128]. ...
In this paper, we want to refute the notion that the microbiota should be considered an organ, given that an organ comprises tissue of similar or different embryological origin, while the microbiota is a pool of different microbial species originating individually from single replications and not from a common ancestral cellular element. Hence, we would like to propose a new morphological interpretation of its nature, based on the comprehensive context in which these microbes live: a muco-microbiotic layer of hollow organs, such as the airways and the bowel. The above concept should represent not only a new terminological annotation but also a more accurate portrayal of the physiology and pathophysiology of these organs. Indeed, a better understanding of the biological nature of this part of the human body can help scientists develop more specific experimental protocols, potentially leading to the establishment of better therapeutic strategies.
