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MSLN knockdown reduces soft agar colony formation and tumor sphere formation. a Western blot of MSLN in several stable knockdown clones by shMSLN RNA. b Proliferation of control (shC) and MSLN knockdown (shMSLN) H2052 and H460 cells, means ± s.d. (n = 6), *P < 0.05. c Representative images and d quantification of soft agar colony formation of shC and shMSLN H2052 and H460 cells. e Representative images and f quantification of tumor sphere formation of shC and shMSLN H2052 and H460 cells. *P < 0.05 vs shC, n = 6
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Background
Lung cancer and pleural mesothelioma are two of the most deadly forms of cancer. The prognosis of lung cancer and mesothelioma is extremely poor due to limited treatment modalities and lack of understanding of the disease mechanisms. We have identified mesothelin as a potentially unique therapeutic target that as a specific advantage app...
Contexts in source publication
Context 1
... pairs of human lung cancer samples with adjacent nor- mal tissue controls were examined by Western blot analysis. MSLN protein levels were not detectable in any of the normal tissues, but clearly in 5 out of 10 lung tumor tissues, i.e. sample pair #1, 3, 5, 9, and 10 in Fig. 1a (H2052). When compared to normal (non-can- cer) human lung epithelial cell line (BEAS-2B) and mesothelial cell line (MeT 5A), expression of MSLN was highly elevated in the cancer cell lines (Fig. 1c and d), suggesting a carcinogenic role of MSLN in lung cancer and mesothelioma. ...
Context 2
... lung tumor tissues, i.e. sample pair #1, 3, 5, 9, and 10 in Fig. 1a (H2052). When compared to normal (non-can- cer) human lung epithelial cell line (BEAS-2B) and mesothelial cell line (MeT 5A), expression of MSLN was highly elevated in the cancer cell lines (Fig. 1c and d), suggesting a carcinogenic role of MSLN in lung cancer and mesothelioma. ( Fig. 2e and f ). These results strongly support the pro- carcinogenic role of MSLN in the tested cell system. Since tumor sphere formation commonly serves as one of the indicators of cancer stem-like cell (CSC) forma- tion, the results of this study also suggest the role of MSLN in CSC regulation, which is supported by our subsequent studies. ...
Context 3
... with previous studies, our presented results indicate an up- regulation of MSLN in human lung tumor tissues and in lung carcinoma and mesothelioma cell lines. Knock- down of MSLN in these cell lines reversed their malig- nant phenotype as indicated by soft agar colony formation, tumor sphere formation, and cell migration and invasion assays (Figs. 2 and 3) as well as tumor formation in animals (Fig. 5). In non-cancerous cells, overexpression of MSLN promoted a malignant pheno- type as indicated by anchorage-independent growth and cell migration and invasion assays (Fig. 4). ...
Context 4
... effect of MSLN on EMT may be cell-line dependent since a previous study by Wang et al. [11] showed that knockdown of MSLN in H2373 mesothelioma cell line did not affect E-cadherin expres- sion but decreased β-catenin expression and increased Slug expression. Our morphological and functional assays confirmed that knockdown of MSLN in H2052 and H460 cells reversed their EMT phenotypes (Figs. 2 and 3), consistent with our EMT markers expression data (Fig. 6). ...
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Citations
... The surface of healthy mesothelial cells of the pleura, pericardium, and peritoneum normally express MSLN in limited amounts [105]. The physiological function of MSLN expression in healthy tissues is little known [106] and it is supposed to be implicated in tumorigenesis, metastasis, and chemoresistance [99,107,108]. MSLN is initially expressed at the cell surface as a precursor protein of 71 kDa; the endoprotease Furin subsequently cleaves it, causing the release of megakaryocyte potentiating factor (MPF), which is a 31 kDa protein, and leaving MSLN in its mature form. Surface MSLN can also be released from the cell membrane by proteases, resulting in a soluble mesothelin-related peptide (SMRP) [109]. ...
Simple Summary
The search for precision medicine applications in mesotheliomas (MM) is taking its first steps. After platinum and pemetrexed chemotherapy, the treatment for relapsed MM remains an unmet clinical need, and the prognosis of MM remains poor even if the recent introduction of immunotherapy. It is known that MM is mainly characterized by inactivanting tumor suppressor alterations and that these, along with some cellular targets or metabolic enzymes, could be potentially amenable to specific therapies The purpose of this review is to take a comprehensive excursus of the main targets and the related evidence regarding possible treatment activities intended for them.
Abstract
Mesothelioma (MM) is an aggressive and lethal disease with few therapeutic opportunities. Platinum-pemetrexed chemotherapy is the backbone of first-line treatment for MM. The introduction of immunotherapy (IO) has been the only novelty of the last decades, allowing an increase in survival compared to standard chemotherapy (CT). However, IO is not approved for epithelioid histology in many countries. Therefore, therapy for relapsed MM remains an unmet clinical need, and the prognosis of MM remains poor, with an average survival of only 18 months. Increasing evidence reveals MM complexity and heterogeneity, of which histological classification fails to explain. Thus, scientific focus on possibly new molecular markers or cellular targets is increasing, together with the search for target therapies directed towards them. The molecular landscape of MM is characterized by inactivating tumor suppressor alterations, the most common of which is found in CDKN2A, BAP1, MTAP, and NF2. In addition, cellular targets such as mesothelin or metabolic enzymes such as ASS1 could be potentially amenable to specific therapies. This review examines the major targets and relative attempts of therapeutic approaches to provide an overview of the potential prospects for treating this rare neoplasm.
... MSLN is a cell surface antigen associated with tumor invasion and is highly expressed in a variety of cancers, acting as a cell adhesion protein. MSLN plays an important role in lung carcinogenesis and epithelial-mesenchymal transition [51]. Targeting MSLN is 11 (which was not certified by peer review) is the author/funder. ...
Understanding disease progression is crucial for detecting critical transitions and finding trigger molecules, facilitating early diagnosis interventions. However, the high dimensionality of data and the lack of aligned samples across disease stages have posed challenges in addressing these tasks. We present a novel framework, Gaussian Graphical Optimal Transport (GGOT), for analyzing disease progressions. The proposed GGOT uses Gaussian graphical models, incorporating protein interaction networks, to characterize the data distributions at different disease stages. Then we use population-level optimal transport to calculate the Wasserstein distances and transport maps between stages, enabling us to detect critical transitions. By analyzing the per-molecule transport distance, we quantify the importance of each molecule and identify trigger molecules. Moreover, GGOT predicts the occurrence of critical transitions in unseen samples and visualizes the disease progression process. We apply GGOT to the simulation dataset and six disease datasets with varying disease progression rates, to show its effectiveness for detecting critical transitions and identifying trigger molecules.
... Targeted therapies against MSLN have showed antitumor activaties in MSLN-positive tumors [16,17]. In lung cancer, MSLN promotes epithelial-mesenchymal transition (EMT) and stemness of tumor cells, tumorwhich may facilitate the occurrence of BM [18]. Recently, MSLN-specific cellular immune responses were identified in the blood of patients with BM and regarded as a predictor for survival, which indicates the involvement of MSLN in BM [19]. ...
... MSLN, a cell surface glycoprotein, is highly expressed in various tumor tissues [18,28,29,40,41], while it is found at very low levels in normal human tissues [10,42]. ...
... A study showed that knockdown of MSLN significantly inhibits in vitro cell adhesion, migration, and invasion (critical steps necessary for metastasis), and also reverses EMT and attenuates stem cell properties in lung cancer cells [18]. In this study, we found that MSLN is not only a specific expression of the tumor antigen in situ in lung cancer compared to paracancer, but its expression is further elevated in brain metastases. ...
Background
Brain metastasis (BM) is common among cases of advanced non-small cell lung cancer (NSCLC) and is the leading cause of death for these patients. Mesothelin (MSLN), a tumor-associated antigen expressed in many solid tumors, has been reported to be involved in the progression of multiple tumors. However, its potential involvement in BM of NSCLC and the underlying mechanism remain unknown.
Methods
The expression of MSLN was validated in clinical tissue and serum samples using immunohistochemistry and enzyme-linked immunosorbent assay. The ability of NSCLC cells to penetrate the blood-brain barrier (BBB) was examined using an in vitro Transwell model and an ex vivo multi-organ microfluidic bionic chip. Immunofluorescence staining and western blotting were used to detect the disruption of tight junctions. In vivo BBB leakiness assay was performed to assess the barrier integrity. MET expression and activation was detected by western blotting. The therapeutic efficacy of drugs targeting MSLN (anetumab) and MET (crizotinib/capmatinib) on BM was evaluated in animal studies.
Results
MSLN expression was significantly elevated in both serum and tumor tissue samples from NSCLC patients with BM and correlated with a poor clinical prognosis. MSLN significantly enhanced the brain metastatic abilities of NSCLC cells, especially BBB extravasation. Mechanistically, MSLN facilitated the expression and activation of MET through the c-Jun N-terminal kinase (JNK) signaling pathway, which allowed tumor cells to disrupt tight junctions and the integrity of the BBB and thereby penetrate the barrier. Drugs targeting MSLN (anetumab) and MET (crizotinib/capmatinib) effectively blocked the development of BM and prolonged the survival of mice.
Conclusions
Our results demonstrate that MSLN plays a critical role in BM of NSCLC by modulating the JNK/MET signaling network and thus, provides a potential novel therapeutic target for preventing BM in NSCLC patients.
... Also, MSLN expression in tumors was shown to be associated with a signi cant increase in microvessel density in newly formed pancreatic metastatic tissue [36]. Another study demonstrated that MSLN knockdown signi cantly inhibits in vitro cell adhesion, migration, and invasion (critical steps necessary for metastasis) and also reverses EMT and attenuates stem cell properties among cancer cells [30]. ...
Brain metastasis (BM) is common among cases of advanced non-small cell lung cancer (NSCLC) and is the leading cause of death for these patients. Mesothelin (MSLN), a tumor-associated antigen expressed in many solid tumors, has been reported to be involved in the progression of multiple tumors. However, its potential involvement in BM of NSCLC and the underlying mechanism remain unknown. In this study, we found that MSLN expression was significantly elevated in both serum and tumor tissue samples from NSCLC patients with BM and correlated with a poor clinical prognosis. Our in vitro and ex vivo experiments demonstrated that MSLN significantly enhanced the brain metastatic abilities of NSCLC cells, especially blood–brain barrier (BBB) extravasation. Mechanistically, we found that MSLN facilitated the expression and activation of MET through the c-Jun N-terminal kinase (JNK) signaling pathway, which allowed tumor cells to disrupt tight junctions and the integrity of the BBB and thereby penetrate the barrier. Intriguingly, our in vivo experiments indicated that drugs targeting MSLN (anetumab) and MET (crizotinib) effectively blocked the development of BM and prolonged the survival of mice, with anetumab showing a superior therapeutic effect compared with crizotinib. In conclusion, our results demonstrate that MSLN plays a critical role in BM of NSCLC by modulating the JNK/MET signaling network and thus, provides a potential novel therapeutic target for preventing BM in NSCLC patients.
... Further studies have shown that MSLN promotes tumor cell survival and proliferation through NF-kB pathway activation, resulting in an increase of interleukin-6 level (26). Recently, He and colleagues demonstrated that MSLN control tumorigenicity and metastatic potential through epithelial-tomesenchymal transition and stem properties of mesothelioma cell lines (27). Interestingly, genetic knockdown of MSLN significantly reversed EMT and attenuated stem cell properties, in addition to inhibiting tumor growth and metastasis; ectopic overexpression of MSLN induced the malignant phenotype of non-tumoral cells, supporting its role as an oncogene (27). ...
... Recently, He and colleagues demonstrated that MSLN control tumorigenicity and metastatic potential through epithelial-tomesenchymal transition and stem properties of mesothelioma cell lines (27). Interestingly, genetic knockdown of MSLN significantly reversed EMT and attenuated stem cell properties, in addition to inhibiting tumor growth and metastasis; ectopic overexpression of MSLN induced the malignant phenotype of non-tumoral cells, supporting its role as an oncogene (27). However, the tumor model in these studies does not resemble the immune-matrix tumor microenvironment (TME) of human mesothelioma. ...
Background
The combination of immunobiological agents with immune checkpoint proteins is a promising treatment for malignant pleural mesothelioma (MPM). Mesothelin and anti-PD-L1 antibody-drug conjugates specifically target malignant neoplastic cells, inhibit the migration and invasion of neoplastic cells, and restore the immune landscape. In this study, we confirmed the importance of mesothelin and examined the relationship between mesothelin and the immune landscape of the tumor microenvironment (TME) in two MPM cohorts.
Methods
The discovery cohort included 82 MPM cases. Tissue microarray slides were generated, and samples were processed for hematoxylin & eosin staining, immunohistochemistry, and immunofluorescence assays. The relationship between mesothelin, biomarkers of histogenesis, histological aggressiveness, PD-L1, immune cells (CD4, CD8, CD20, CD68), and collagen type I and type V fibers was evaluated by quantitative digital analyses. The outcome was the survival time until death from disease recurrence. The exploratory cohort included 87 malignant mesothelioma (MESO) patients from The Cancer Genome Atlas database.
Results
Most patients were male (70.7%) with a history of asbestos exposure (53.7%) and with the epithelioid subtype (89%). Surgical resection was performed in 85.4% of patients, and 14.6% received chemotherapy; 59.8% of patients died from disease extension to the mediastinum. Low tumor mesothelin expression was associated with tumor necrosis and nuclear grade 1, whereas high mesothelin expression was significantly associated with the epithelioid histotype and high density of T cells CD8+, macrophages CD68+, and collagen type I fibers. Cox multivariate analysis showed a high risk of death for non-operated patients [hazard ratio (HR), 3.42 (1.15–10.16)] with low tumor mesothelin levels [HR, 2.58 (1.09–6.10)] and high PD-L1 and low infiltration of T cells CD4+ [HR, 3.81 (1.58–9.18)]. In the exploratory cohort, low mesothelin and high COL1A1 and COL5A1 expression were associated with poor overall survival.
Conclusion
Tumor mesothelin expression associated with the TME immune landscape predicts the risk of death for patients with MPM and could be a new target for immunotherapy in MPM.
... Epithelial-mesenchymal transition (EMT) is the process by which epithelial cells lose their cell polarity and ability to adhere to other cells, and gain migratory and invasive properties, becoming mesenchymal stem cells (11). The effects of EMT have been investigated (12,13). EMT serves an important role in tumor metastasis and recurrence, as it breaks down cell-cell and cell-extracellular matrix connections to facilitate cancer cell migration through the extracellular matrix. ...
Suppressor of cytokine signaling 2 (SOCS2) plays an essential role in a number of physiological phenomena and functions as a tumor suppressor. Understanding the predictive effects of SOCS2 on non-small cell lung cancer (NSCLC) is urgently needed. The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases were used to assess SOCS2 gene expression levels in NSCLC. The clinical significance of SOCS2 was evaluated through Kaplan-Meier curve analysis and the analysis of related clinical factors. Gene Set Enrichment Analysis (GSEA) was used to identify the biological functions of SOCS2. Subsequently proliferation, wound-healing, colony formation and Transwell assays, and carboplatin drug experiments were used for verification. The results revealed that SOCS2 expression was low in the NSCLC tissues of patients in TCGA and GEO database analyses. Downregulated SOCS2 was associated with poor prognosis, as determined by Kaplan-Meier survival analysis (HR 0.61, 95% CI 0.52-0.73; P<0.001). GSEA showed that SOCS2 was involved in intracellular reactions, including epithelial-mesenchymal transition (EMT). Cell experiments indicated that knockdown of SOCS2 caused the malignant progression of NSCLC cell lines. Furthermore, the drug experiment showed that silencing of SOCS2 promoted the resistance of NSCLC cells to carboplatin. In conclusion, low expression of SOCS2 was associated with poor clinical prognosis by effecting EMT and causing drug resistance in NSCLC cell lines. Furthermore, SOCS2 could act as a predictive indicator for NSCLC.
... Targeting invasion in the treatment of mesothelioma is of great interest, given the central role of invasion in the morbidity of this disease. Previous clinical studies have attempted to inhibit invasion in mesothelioma patients by targeting mesothelin [34, 35], a membrane-bound protein that stimulates anchorage-independent growth, migration, and invasion [36,37]. However, progression free survival was not different between the treatment and control groups. ...
Mesothelioma is characterised by its aggressive invasive behaviour, affecting the surrounding tissues of the pleura or peritoneum. We compared an invasive pleural model with a non-invasive subcutaneous model of mesothelioma and performed transcriptomic analyses on the tumour samples. Invasive pleural tumours were characterised by a transcriptomic signature enriched for genes associated with MEF2C and MYOCD signaling, muscle differentiation and myogenesis. Further analysis using the CMap and LINCS databases identified geldanamycin as a potential antagonist of this signature, so we evaluated its potential in vitro and in vivo. Nanomolar concentrations of geldanamycin significantly reduced cell growth, invasion, and migration in vitro. However, administration of geldanamycin in vivo did not result in significant anti-cancer activity. Our findings show that myogenesis and muscle differentiation pathways are upregulated in pleural mesothelioma which may be related to the invasive behaviour. However, geldanamycin as a single agent does not appear to be a viable treatment for mesothelioma.
... However, the physiological function of MSLN remains unknown in gastric cancer patho-physiology. MSLN is a cell surface differentiation antigen, which is an attractive anticancer therapeutic target as its normal expression is largely limited to the cells of the pleura, pericardium, and peritoneum [16,17]. Moreover, MSLN knockout mice have no observable phenotype [18,19]. ...
... The AGS cell line overexpressing MSLN demonstrated increased proliferation, motility and invasion. In this line, the impact of MSLN on several EMT and CSC regulatory genes studied by Xiaoqing He et al. suggests that MSLN may act as a master regulator of EMT that controls both local invasion and distant metastasis [16]. ...
Mesothelin (MSLN), a tumor-associated antigen, is upregulated in various malignancies, including gastric cancer (GC). In addition, MSLN is found in the blood-stream of affected individuals, where it is referred to as soluble MSLN-related protein (SMRP). This study aims to investigate the role of MSLN in GC and evaluate its potential as a plasma biomarker for diagnosis and prognosis. Toward that end, GC tissues were obtained, upon signed consent, from affected individuals undergoing surgery or endoscopy (n = 82). Quantitative RT-PCR and immunohistochemistry were performed to determine MSLN expression. Simultaneously, The Cancer Genome Atlas (TCGA) database was mined to evaluate global status of MSLN gene expression in gastric cancer. Next, in vitro cell-culture studies were conducted to evaluate MSLN-driven proliferation properties. Using ELISA, sera from 55 GC-affected individuals were tested for MSLN level. Additionally, plasma mesothelin levels were compared in 6 cases before and after surgery. Upregulated MSLN expression was found in GC tissues, compared to adjacent normal tissues (p < 0.001). Cell culture studies with a MSLN-overexpressing stable GC line showed increased cell proliferation and invasion with ectopic MSLN. Additionally, gene-set-enrichment-analysis (GSEA) revealed an association of MSLN with the genes involved in the epithelial-mesenchymal transition and G2/M checkpoint. GC-affected cases showed higher serum MSLN levels, compared to healthy controls, with rapid decrease post-surgery. We found that MSLN upregulation correlates with poor clinical outcome and promotes growth advantage to GC cells in vitro. With further experimental evidences, we propose that MSLN could potentially be used as a plasma biomarker for diagnosis of GC.
... The role of Amotl2 in mOSCC may more closely align with that in mammary epithelial cells, but that remains an area for further exploration. Similarly, we also observed that high p63 expression was associated with the downregulation of Msln, which encodes mesothelin, a protein that promotes EMT and invasion in breast, lung, and ovarian cancers [84][85][86]. A similar dichotomy of p63 function in cell migration and invasion has been reported before and has been attributed to an oscillatory expression of the ∆Np63 isoform that results in its tumor suppressive activities [25]. ...
Simple Summary
Oral squamous cell carcinomas (OSCC) are the most common malignancies affecting the oral cavity and account for 40% of all head and neck squamous cell carcinoma cases. Unfortunately, patient outcomes are generally unfavorable, as diagnosis normally occurs at a late stage of the disease, as well as a lack of effective targeted treatments. Our identification of potential prognostic biomarkers is of particular importance to the field and could lead to the generation of effective targeted therapies which may improve patient outcomes and survival. Here, we utilize a mouse model of OSCC to explore the role of p63 as an important oncogenic transcription factor in the control of OSCC proliferation and migration. We also generate a p63-driven gene expression signature for mouse OSCC which identifies both novel and conserved genes and pathways, which may be relevant in human disease and which may serve as potential biomarkers and targets for future therapeutics.
Abstract
Oral squamous cell carcinoma (OSCC) is the most common malignancy of the oral cavity and is linked to tobacco exposure, alcohol consumption, and human papillomavirus infection. Despite therapeutic advances, a lack of molecular understanding of disease etiology, and delayed diagnoses continue to negatively affect survival. The identification of oncogenic drivers and prognostic biomarkers by leveraging bulk and single-cell RNA-sequencing datasets of OSCC can lead to more targeted therapies and improved patient outcomes. However, the generation, analysis, and continued utilization of additional genetic and genomic tools are warranted. Tobacco-induced OSCC can be modeled in mice via 4-nitroquinoline 1-oxide (4NQO), which generates a spectrum of neoplastic lesions mimicking human OSCC and upregulates the oncogenic master transcription factor p63. Here, we molecularly characterized established mouse 4NQO treatment-derived OSCC cell lines and utilized RNA and chromatin immunoprecipitation-sequencing to uncover the global p63 gene regulatory and signaling network. We integrated our p63 datasets with published bulk and single-cell RNA-sequencing of mouse 4NQO-treated tongue and esophageal tumors, respectively, to generate a p63-driven gene signature that sheds new light on the role of p63 in murine OSCC. Our analyses reveal known and novel players, such as COTL1, that are regulated by p63 and influence various oncogenic processes, including metastasis. The identification of new sets of potential biomarkers and pathways, some of which are functionally conserved in human OSCC and can prognosticate patient survival, offers new avenues for future mechanistic studies.
... 32 There are several diseases along with cancers caused by the IL-17A gene, such as multiple sclerosis, 33 psoriasis, 34 Crohn's disease and ulcerative colitis, 35 systemic lupus erythematosus, 36 lung diseases. 37 It also plays potential role in the formation of stomach cancer, 38,39 colorectal cancer, 40,41 cervical cancer, 42 lung carcinoma, 43 breast carcinoma, 44 and bladder cancer. 45 There are a few case-control studies till now have been investigated to find out the association of human IL-17A gene polymorphisms (rs10484879 and rs3748067) with the susceptibility of CRC. ...
Objective
Interleukin-17A ( IL-17A) genetic polymorphisms are associated with multiple cancer types, including colorectal cancer (CRC). However, no previous study was performed in the Bangladeshi population to evaluate the association. Our study aimed to find the association between two IL-17A variants (rs10484879 C/A and rs3748067 G/A) and susceptibility of CRC.
Methods and Materials
This retrospective case-control study comprised 292 CRC patients and 288 age, sex, and BMI matched healthy volunteers. Genotyping of both variants was done by the tetra-primer ARMS-PCR method, and the results were analyzed by the SPSS software package (version-25.0).
Results
Logistic regression analysis indicated that in case of IL-17A rs10484879 polymorphism, AC and AA genotype carriers showed 2.44- and 3.27-times significantly increased risk for CRC development (OR = 2.44, P = .0008 and OR = 3.27, P = .0133, individually). A significant association was also observed for AC + AA genotype (OR = 2.58, P = .0001). Again, over-dominant and allelic model revealed statistically significant link to CRC risk (OR = 2.13, P = .0035 and OR = 2.22, P = .001). For rs3748067 polymorphism, AG and AA genotype carriers showed 2.30- and 2.45-times enhanced risk for CRC (OR = 2.30, P = .005 and OR = 2.45, P = .031). A statistically significant association was also observed for AG + AA genotype (OR = 2.35, P = .001), over-dominant model (OR = 2.05, P = .014), and allelic model (OR = 2.11, P = .0004).
Conclusion
This study highlights that IL-17A rs10484879 and rs3748067 polymorphisms may be associated with CRC development. However, further functional research with larger samples may reveal more statistically significant outcomes.
