61 research outputs found
Mineralogy as useful tool for medicine: the study of hazardous materials in biological tissues
No full textSera from Patients with Malignant Pleural Mesothelioma Tested Positive for IgG Antibodies against SV40 Large T Antigen: The Viral Oncoprotein
No full textMalignant pleural mesothelioma (MPM), a fatal tumor, is mainly linked to the asbestos exposure. It has been reported that together with the inhalation of asbestos fibers, other factors are involved in the MPM onset, including simian virus 40 (SV40). SV40, a polyomavirus with oncogenic potential, induces (i) in vitro the mesenchymal cell transformation, whereas (ii) in vivo the MPM onset in experimental animals. The association between MPM and SV40 in humans remains to be elucidated. Sera (n = 415) from MPM-affected patients (MPM cohort 1; n = 152) and healthy subjects (HSs, n = 263) were investigated for their immunoglobulin G (IgG) against simian virus 40 large tumor antigen (Tag), which is the transforming protein. Sera were investigated with an indirect enzyme-linked immunosorbent assay (ELISA) using two synthetic peptides from SV40 Tag protein. SV40 Tag protein was evaluated by immunohistochemical (IHC) staining on MPM samples (MPM cohort 2; n = 20). Formalin-fixed and paraffin-embedded (FFPE) samples were obtained from MPM patients unrelated to MPM serum donors. The proportion of sera, from MPM patients, showing antibodies against SV40 Tag (34%) was significantly higher compared to HSs (20%) (odds ratio 2.049, CI 95% 1.32–3.224; p=0.0026). Immunohistochemical staining (IHS) assays showed SV40 Tag expression in 8/20, 40% of MPM specimens. These results indicate that SV40 is linked to a large fraction of MPM. It is worth noting that the prevalence of SV40 Tag antibodies detected in sera from cohort 1 of MPM patients is similar to the prevalence of SV40 Tag found to be expressed in FFPE tissues from MPM cohort 2
Characterization of human malignant mesothelioma cell lines orthotopically implanted in the pleural cavity of immunodeficient mice for their ability to grow and form metastasis
No full textAbstract Background Malignant pleural mesothelioma (MPM) is a tumor known to be resistant to conventional therapies. Thus, an in vivo model can represent an important tool for assessing the efficacy of novel approaches in the treatment of MPM. Presently, human MPM cells have been grown orthotopically in mice upon transplantation of tumor masses or tumor cell suspensions following surgery. In these models however, surgery can interfere with the tumor growth and the early stages of tumor development cannot be easily explored. Finally, results may not be so accurate due to implantation of potentially different tumor samples in different experimental groups. Our work aimed at establishing a nude mouse model xenotransplanted with human MPM cell lines in which tumor progression exhibits some features of the human disease. Methods Three distinct human MPM cell lines previously established from MPM patients displaying two different phenotypes, biphasic (MM-B1 and IST-Mes3) and epithelioid (IST-Mes2), were directly injected into the pleural cavity of nude mice. At different times, mice were sacrificed for autopsy, tumor nodules were counted and then removed for histology. Presence of metastases in visceral organs was also monitored. Results IST-Mes2 cells were unable to grow in nude mice. MM-B1 and IST-Mes3 cells were capable of growing in nude mice and formed tumor nodules in the pleura. Post-mortem examination showed that MPM cells progressively colonized the parietal and visceral pleura, the diaphragm, the mediastinum and, lastly the lung parenchyma. No pneumo-thorax was evidenced in the mice. Pleural effusions as well as lymph node metastases were observed only at later times. Conclusion This model mimics the progression of human malignant mesothelioma and it is easy to perform and reproducible; therefore it can be useful to study human MPM biology and evaluate the efficacy of novel therapies.</p
Environmental Scanning Electron Microscopy applied to the identification of asbestos fibers in histological sections
No full textDifferential Diagnosis of Malignant Pleural Mesothelioma on Cytology: A Gene Expression Panel Versus BRCA1-Associated Protein 1 and p16 Tests
No full textPleural effusions are among the first clinical manifestations of malignant pleural mesothelioma (MPM) and often constitute the only available material for diagnosis. Although an MPM diagnosis can be reliable on cytology, the reported sensitivity is low (30% to 75%). Particularly, it can be hard to discriminate epithelioid MPM, the most common histotype, from reactive mesothelial hyperplasia (MH). Currently, BRCA1-associated protein 1 (BAP1) and CDKN2A (p16), evaluated by immunohistochemistry and fluorescent in situ hybridization, respectively, are the most valuable markers to discriminate MPM and MH. Both markers have a high specificity, but their sensitivity is not always satisfying, even when used together. We have recently developed a 117-gene expression panel, based on Nanostring technology, able to differentiate epithelioid MPM from MH pleural tissues better than BAP1 and p16. Herein, we evaluated the efficacy of the same panel on an independent retrospective cohort of 23 MPM and 11 MH pleural effusions (cell blocks and smears). The overall sensitivity and specificity of the panel were equal to 0.9565 and 1, respectively. Moreover, the panel performance was compared with BAP1 and p16 on 25 cell blocks. Sensitivity levels of gene panel, BAP1 alone, p16 alone, and BAP1 plus p16 were 1, 0.5882, 0.4706, and 0.7647, respectively. Specificity was always 1. Although further validation is needed, this gene panel could really facilitate patients' management, allowing a definitive MPM diagnosis directly on pleural effusions
Uso del microRNA hsa-miR-197-3p come marcatore di esposizione all’amianto e di mesotelioma maligno della pleura
No full textSi descrive l'uso del microRNA hsa -miR-197-3p come biomarcatore per la valutazione dell'esposizione all'amianto, del livello di esposizione nei lavoratori ex-esposti all'amianto, l'insorgenza e lo sviluppo del mesotelioma maligno della pleura e l'identificazione dei diversi istotipi del mesotelioma maligno della pleura, relativi metodi per la diagnosi del mesotelioma pleurico maligno e per l'identificazione degli istotipi del mesotelioma pleurico maligno, nonché relativo kit
Circulating microRNA-197-3p as a potential biomarker for asbestos exposure
No full textAsbestos is considered the main cause of diseases in workers exposed to this mineral in the workplace as well as an environmental pollutant. The association between asbestos and the onset of different diseases has been reported, but asbestos exposure specific biomarkers are not known. MicroRNAs (miRNAs) are small, single-strand, non-coding RNAs, with potential value as diagnostic, prognostic, and predictive markers in liquid biopsies. Sera collected from workers ex-exposed to asbestos (WEA) fibers were compared with sera from healthy subjects (HS) of similar age, as liquid biopsies. The expression of the circulating miRNA 197-3p was investigated employing two different highly analytical PCR methods, i.e. RT-PCR and dd-PCR. MiR-197-3p levels were tested in sera from WEA compared to HS. MiR-197-3p tested dysregulated in sera from WEA (n=75) compared to HS (n=62). Indeed, miR-197-3p was found to be 2.6 times down-regulated in WEA vs. HS (p=0.0001***). In addition, an inverse correlation was detected between miR-197-3p expression level and cumulative asbestos exposure, being this miRNA down-regulated 2.1 times in WEA, with high cumulative asbestos exposure, compared to WEA with low exposure (p=0.0303*). Circulating miR-197-3p, found to be down regulated in sera from WEA, is proposed as a new potential biomarker of asbestos exposure
Nicotinamide phosphoribosyltransferase (NAMPT) is over-expressed and abundantly released from malignant pleural mesothelioma cells becoming a potential biomarker.
No full textIntroduction
Malignant pleural mesothelioma (MPM) is an aggressive and incurable cancer of the pleural
surface. Chronic inflammation, oxidative stress, and persistent aberrant signaling due to asbestos
exposure led to mesothelial cells transformation over years. Despite increasing studies on MPM
biology, continue efforts to identify novel biomarkers and tumor vulnerabilities to be targeted are
needed. Nicotinamide adenine dinucleotide (NAD) biosynthesis is essential to support tumor
energetic needs, as well as to regulate NADPH-mediated detoxification system. Activation of NAD
metabolism through its rate-limiting biosynthetic enzyme nicotinamide phosphoribosyltransferase
(NAMPT) has been identified as key event to support cancer metabolic rewiring. NAMPT, in
addition to possess a key function in NAD generation, can be secreted in the extracellular space
(eNAMPT), where it behaves as a mediator of inflammation, regulating tumor-host interactions.
NAD/NAMPT axis emerges deregulated in several tumors; however, no data are available in MPM.
Methods
NAMPT-NAD axis (NAD-biosynthetic enzymes expression, activities, and metabolites) in MPM
cell lines and primary samples was studied using biochemical, enzymatic, immunochemical assays.
eNAMPT was evaluated exploiting commercial ELISA assay in sera and pleural effusions for more
than 100 MPM patients with different histotype derived from AOU-Alessandria Biobank. In silico
analysis using TCGA database was performed to correlate NAMPT expression and biological
processes analyzing the cohort of MPM patients.
Results
Bioinformatics analysis on TCGA database showed that NAMPT is the main expressed NAD-
biosynthetic enzymes in MPM, and its expression correlates with hallmark gene sets related to
inflammation, metabolic and signaling pathways. RT-PCR and western blot analysis on MPM cell
lines and mesothelioma primary cells vs normal mesothelium (Met-5A, or primary mesothelial
cells) confirmed that NAMPT is overexpressed in MPM. Data showed that NAD levels are similar
in the comparison between tumor and normal tissue, while NADP levels are increased in MPM cell
lines, suggesting an impact on cellular redox state and metabolic homeostasis that will further
evaluate. Within the extracellular space, we revealed significantly increased serum eNAMPT levels
from a cohort of 115 MPM patients compared to healthy donors. eNAMPT is strongly released in
pleural effusions from the same MPM patients, mainly in MPM with the most aggressive
sarcomatoid phenotype. Moreover, we found also a second biosynthetic enzyme nicotinate
phosphoribosyltransferase (NAPRT) released by MPM cells. Lastly, preliminary data showed that
MPM cells are uniquely sensitive to NAMPT inhibition.
Conclusions
Overall, these data support the hypothesis of an impact of NAD/NAMPT axis in MPM biology and
highlight a potential role of eNAMPT as biomarker with a functional activity like a damage-
associated molecular pattern (DAMPs)/cytokine in MPM that will be further investigated
Nicotinamide phosphoribosyltransferase (NAMPT) is over-expressed and abundantly released from malignant pleural mesothelioma cells becoming a potential biomarker.
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