Expression of Cartilage Oligomeric Matrix Protein in colorectal cancer is an adverse prognostic factor and correlates negatively with infiltrating immune cells and PD-L1 expression

IntroductionCartilage Oligomeric Matrix Protein (COMP) is an oncogenic protein that has been associated with a decrease in infiltrating T-cells in periampullary adenocarcinoma. This study aimed to investigate whether this is also the case for colorectal cancer (CRC) and to evaluate the relationship between COMP expression and clinopathological features.MethodsImmunohistochemistry was used to determine the expression levels of COMP in tumor cells and stroma in primary tumors from a cohort of 537 CRC patients. The expression of immune cell markers, including CD3+, CD8+, FoxP3+, CD68+, CD56+, CD163+, and PD-L1, was evaluated previously. Tumor fibrosis was assessed by Sirius Red staining and evaluation of collagen fiber organization.ResultsCOMP expression correlated positively with TNM-stage and grade of differentiation. Patients with CRC expressing high levels of COMP had significantly shorter OS than those with low COMP expression (p<0.0001), and fewer infiltrating T-cells were detected in tumors with high COMP expression. Additionally, a negative correlation was identified between the expression of COMP and PD-L1 on both tumor cells and immune cells. Cox regression analysis showed that tumors expressing high levels of COMP had significantly shorter OS, independent of all evaluated immune cell markers. Tumor fibrosis was correlated with high expression of COMP in the stroma (p<0.0001), and tumors with high levels of COMP expression and denser fibrosis displayed more sparse immune cell infiltration.DiscussionThe results suggest that COMP expression in CRC may exert an immune regulatory effect by increasing dense fibrosis and decreasing immune cell infiltration. These findings support the notion that COMP is an important factor in the development and progression of CRC

Complement inhibitor CSMD1 acts as tumor suppressor in human breast cancer

Human CUB and Sushi multiple domains 1 (CSMD1) is a membrane-bound complement inhibitor suggested to act as a putative tumor suppressor gene, since allelic loss of this region encompassing 8p23 including CSMD1 characterizes various malignancies. Here, we assessed the role of CSMD1 as a tumor suppressor gene in the development of breast cancer in vitro and in vivo. We found that human breast tumor tissues expressed CSMD1 at lower levels compared to that in normal mammary tissues. The decreased expression of CSMD1 was linked to a shorter overall survival of breast cancer patients. We also revealed that expression of CSMD1 in human breast cancer cells BT-20 and MDA-MB-231 significantly inhibited their malignant phenotypes, including migration, adhesion and invasion. Conversely, stable silencing of CSMD1 expression in T47D cells enhanced cancer cell migratory, adherent and clonogenic abilities. Moreover, expression of CSMD1 in the highly invasive MDA-MB-231 cells diminished their signaling potential as well as their stem cell-like properties as assessed by measurement of aldehyde dehydrogenase activity. In a xenograft model, expression of CSMD1 blocked the ability of cancer cells to metastasize to secondary sites in vivo, likely via inhibiting local invasion but not the extravasation into distant tissues. Taken together, these findings demonstrate the role of CSMD1 as a tumor suppressor gene in breast cancer

Correction: Complement inhibitor CSMD1 acts as tumor suppressor in human breast cancer

This article has been corrected: In Figure 4B, the image of MDA-MB-231 cells expressing CSMD1 is an accidental duplicate of the image showing invaded BT-20 cells expressing CSMD1 in Figure 4A. The correct Figure 4, produced using the original data, is shown below. The authors declare that these corrections do not change the results or conclusions of this paper. Original article: Oncotarget. 2016; 7:76920–76933. https://doi.org/10.18632/oncotarget.1272

Biochemical and cellular molecular approach of the effect of breast cancer cells in the expression of extracellular matrix molecules and functional properties of endothelial cells

Tumor microenvironment is of outmost importance, considering the fact that cancer cells recruit normal stromal cells, like endothelial cells, in order to facilitate cancer progression.The aim of the present study was to determine the cellular responses that breast cancer cells (BCC) (MDA-MB-231 & MCF-7) elicit in the human umbilical vein endothelial cells (HUVEC). For this purpose two models were utilized; one involves HUVEC culture in the presence of BCC-derived conditioned media (CM) and vice versa, and the other co-culture of both cell populations in a Transwell system, introducing a new factor to all system, that of cell communication. We found that CM from BBCs decreases HUVEC cells migration, affecting also cytoskeleton organization, whereas the adhesion, and invasion of cancer cells is favored by the presence of HUVEC-secreted matrix effectors. In parallel, BBCs migration is up-regulated by the action HUVEC-CM. Such functional interactions were evaluated at the molecular level by the study of the expression of hyaluronan, its receptor CD44, its synthase enzyme HAS2, adhesion molecules ICAM, VCAM, matrix metalloproteases, as well as cell surface syndecans. The expression and activity of the proteasome was also evaluated.Conclusively, breast cancer-endothelium interactions provide regulatory signals facilitating tumor progression. It is therefore concluded that the breast cancer cells secreted effectors are of crucial importance to signal the endothelial cells and to modify their glycocalyx expression and properties. The upregulation of endothelium adhesion molecules, reorganization of the endothelial cytoskeleton as well as breast cancer cells, and up-regulation of proteosomal activity summarize the driving forces of cancer cells to the endothelial in order to disrupt the endothelium creating “holes” with the endothelial cells. These observations will force new studies in the field as to evaluate in detail the underlying signaling mechanisms and open a new area to design novel drugs for pharmacological targeting of breast cancer at the ECM level.Το μικροπεριβάλλον του όγκου είναι ιδιαίτερα σημαντικό, δεδομένου ότι τα καρκινικά κύτταρα στρατολογούν και φυσιολογικά κύτταρα του στρώματος, όπως τα ενδοθηλιακά με σκοπό την εξέλιξη του όγκου.Στο πλαίσιο της μελέτης των παρακρινών αλληλεπιδράσεων μεταξύ καρκινικών και ενδοθηλιακών κυττάρων αναπτύχθηκαν δύο πειραματικά κυτταρικά μοντέλα. Στο πρώτο μοντέλο μελετήθηκε η επίδραση του εθισμένου μέσου καλλιέργειας των καρκινικών κυττάρων στα ενδοθηλιακά κύτταρα και αντίστροφα, ενώ στο δεύτερο εισάγοντας έναν ακόμα παράγοντα στο σύστημα – αυτόν της κυτταρικής επικοινωνίας – πραγματοποιήθηκαν συγκαλλιέργειες ενδοθηλιακών και καρκινικών κυττάρων, έτσι ώστε να υπάρχει η δυνατότητα αλληλεπίδρασής τους και ανταλλαγής διαλυτών παραγόντων. Μεταξύ των αποτελεσμάτων σημειώνεται η σημαντική μείωση της επούλωσης πληγής (κυτταρική κινητικότητα) των ενδοθηλιακών κυττάρων παρουσία εθισμένου μέσου καλλιέργειας καρκινικών κυττάρων, παρέχοντας με αυτόν τον τρόπο στα καρκινικά κύτταρα τις απαραίτητες διόδους για να μεταναστεύσουν. Αντίστοιχα, η κυτταρική μετανάστευση, η προσκόλληση, καθώς και η διηθητική ικανότητα των καρκινικών κυττάρων επάγεται από το ενδοθήλιο. Οι λειτουργικές αυτές επιδράσεις αξιολογήθηκαν περαιτέρω σε μοριακό επίπεδο με τη μελέτη της έκφρασης του υαλουρονικού, του υποδοχέα CD44, της συνθάσης ΗΑS2, των μορίων προσκόλλησης ICAM, VCAM, των μεταλλοεξαρτώμενων πρωτεασών, καθώς και των συνδεκανών της κυτταρικής επιφάνειας. Μελετήθηκε επίσης η έκφραση και η ενεργότητα του πρωτεασώματος. Οι αλλαγές στο προφίλ της γονιδιακής έκφρασης υποδεικνύουν τη σημασία του δυναμικού δικτύου του εξωκυττάριου χώρου και την επίδρασή του στις ιδιότητες των ενδοθηλιακών και καρκινικών κυττάρων. Η αυξορρύθμιση των μορίων προσκόλλησης του ενδοθηλίου, η αναδιοργάνωση του ενδοθηλιακού κυτταροσκελετού και των καρκινικών κυττάρων, καθώς και η επαγωγή της ενεργότητας του πρωτεασώματος αποτελούν τις κινητήριες δυνάμεις της διεπικοινωνίας μεταξύ ενδοθηλίου και καρκινικών κυττάρων που έχουν ως στόχο την διάσπαση του ενδοθηλίου. Τα αποτελέσματα της διατριβής υπογραμμίζουν τη σημαντικότητα των κυτταρικών μιμητικών μοντέλων για την κατανόηση των μηχανισμών διείσδυσης των καρκινικών κυττάρων στο ενδοθήλιο, αποτελούν το έναυσμα για την αξιολόγηση των υποκείμενων σηματοδοτικών μηχανισμών και ανοίγουν νέους δρόμους για τον σχεδιασμό νέων αντικαρκινικών παραγόντων για την φαρμακολογική στόχευση σε επίπεδο εξωκυττάριου χώρου

Evaluation of Glycosaminoglycans Biological Age in Cells and Tissues

Bomb pulse refers to the sharp increase of the atmospheric levels of the carbon isotope 14C due to nuclear testing during the late 1950s and early 1960s. After the nuclear test ban, atmospheric 14C levels decreased exponentially because of the diffusion and equilibration with the ocean and biosphere, as well as the incorporation indirectly into all living cells. Thus, assessing the anthropogenic 14C serves as an isotopic chronometer providing the opportunity to estimate the age of a tissue and its components with a relatively high precision comparing it with the atmospheric levels at a certain time stamp.This protocol describes the use of the bomb pulse dating as a retrospective tracer of tissue components such as the glycosaminoglycans (GAGs) and subsequent proteoglycans turnover using accelerator mass spectrometry

Extracellular matrix: paving the way to the newest trends in atherosclerosis

PURPOSE OF REVIEW: The extracellular matrix (ECM) is critical for all aspects of vascular pathobiology. In vascular disease the balance of its structural components is shifted. In atherosclerotic plaques there is in fact a dynamic battle between stabilizing and proinflammatory responses. This review explores the most recent strides that have been made to detail the active role of the ECM - and its main binding partners - in driving atherosclerotic plaque development and destabilization. RECENT FINDINGS: Proteoglycans-glycosaminoglycans (PGs-GAGs) synthesis and remodelling, as well as elastin synthesis, cross-linking, degradation and its elastokines potentially affect disease progression, providing multiple steps for potential therapeutic intervention and diagnostic targeted imaging. Of note, GAGs biosynthetic enzymes modulate the phenotype of vascular resident and infiltrating cells. In addition, while plaque collagen structure exerts very palpable effects on its immediate surroundings, a new role for collagen is also emerging on a more systemic level as a biomarker for cardiovascular disease as well as a target for selective drug-delivery. SUMMARY: The importance of studying the ECM in atherosclerosis is more and more acknowledged and various systems are being developed to visualize, target and mimic it

Novel potential inhibitors of complement system and their roles in complement regulation and beyond

The complement system resembles a double-edged sword since its activation can either benefit or harm the host. Thus, regulation of this system is of utmost importance and performed by several circulating and membrane-bound complement inhibitors. The pool of well-established regulators has recently been enriched with proteins that either share structural homology to known complement inhibitors such as Sushi domain-containing (SUSD) protein family and Human CUB and Sushi multiple domains (CSMD) families or extracellular matrix (ECM) macromolecules that interact with and modulate complement activity. In this review, we summarize the current knowledge about newly discovered complement inhibitors and discuss their implications in complement regulation, as well as in processes beyond complement regulation such cancer development. Understanding the behavior of these proteins will introduce new mechanisms of complement regulation and may provide new avenues in the development of novel therapies

Extracellular matrix structure.

Extracellular matrix (ECM) is a non-cellular three-dimensional macromolecular network composed of collagens, proteoglycans/glycosaminoglycans, elastin, fibronectin, laminins, and several other glycoproteins. Matrix components bind each other as well as cell to adhesion receptors forming a complex network into which cells reside in all tissues and organs. Cell surface receptors transduce signals into cells from ECM, which regulate diverse cellular functions, such as survival, growth, migration, and differentiation and are vital for maintaining normal homeostasis. ECM is a highly dynamic structural network that continuously undergoes remodeling mediated by several matrix-degrading enzymes during normal and pathological conditions. Deregulation of ECM composition and structure is associated with the development and progression of several pathologic conditions. This article emphasizes in the complex ECM structure as to provide a better understanding of its dynamic structural and functional multipotency. Where relevant, the implication of the various families of ECM macromolecules in health and disease is also presented

Atherosclerotic plaque features relevant to rupture-risk detected by clinical photon-counting CT ex vivo: a proof-of-concept study

Abstract Background To identify subjects with rupture-prone atherosclerotic plaques before thrombotic events occur is an unmet clinical need. Thus, this proof-of-concept study aims to determine which rupture-prone plaque features can be detected using clinically available photon-counting computed tomography (PCCT). Methods In this retrospective study, advanced atherosclerotic plaques (ex vivo, paraffin-embedded) from the Carotid Plaque Imaging Project were scanned by PCCT with reconstructed energy levels (45, 70, 120, 190 keV). Density in HU was measured in 97 regions of interest (ROIs) representing rupture-prone plaque features as demonstrated by histopathology (thrombus, lipid core, necrosis, fibrosis, intraplaque haemorrhage, calcium). The relationship between HU and energy was then assessed using a mixed-effects model for each plaque feature. Results Plaques from five men (age 79 ± 8 [mean ± standard deviation]) were included in the study. Comparing differences in coefficients (b 1diff) of matched ROIs on plaque images obtained by PCCT and histology confirmed that calcium was distinguishable from all other analysed features. Of greater novelty, additional rupture-prone plaque features proved discernible from each other, particularly when comparing haemorrhage with fibrous cap (p = 0.017), lipids (p = 0.003) and necrosis (p = 0.004) and thrombus compared to fibrosis (p = 0.048), fibrous cap (p = 0.028), lipids (p = 0.015) and necrosis (p = 0.017). Conclusions Clinically available PCCT detects not only calcification, but also other rupture-prone features of human carotid plaques ex vivo. Relevance statement Improved atherosclerotic plaque characterisation by photon-counting CT provides the ability to distinguish not only calcium, but also rupture-prone plaque features such as haemorrhage and thrombus. This may potentially improve monitoring and risk stratification of atherosclerotic patients in order to prevent strokes. Key points • CT of atherosclerotic plaques mainly detects calcium. • Many components, such as intra-plaque haemorrhage and lipids, determine increased plaque rupture risk. • Ex vivo carotid plaque photon-counting CT distinguishes haemorrhage and thrombus. • Improved plaque photon-counting CT evaluation may refine risk stratification accuracy to prevent strokes. Graphical Abstrac

Expression of cartilage oligomeric matrix protein in periampullary adenocarcinoma is associated with pancreatobiliary-type morphology, higher levels of fibrosis and immune cell exclusion

Cartilage oligomeric matrix protein (COMP) is an emerging regulator of tumor progression. The aim of this study was to evaluate the expression of COMP in periampullary adenocarcinoma with respect to prognostic value for survival and relapse, levels of fibrosis and infiltrating immune cells. COMP expression was evaluated using immunohistochemistry in primary tumors and subsets of paired lymph node metastases in tissue microarrays including 175 patients with periampullary adenocarcinoma. Collagen content was assessed with Sirius Red-Fast Green staining. High COMP levels were detected in cancer cells and in stroma, in 46% and 57% of the patients, respectively. High COMP expression was strongly associated with more aggressive pancreatobiliary-type (PB-type) compared to intestinal-type tumors (p < .0001). Importantly, high expression of COMP correlated with the exclusion of cytotoxic T-cells from the cancer cell compartment of the tumors, particularly in PB-type tumors. Higher levels of fibrosis measured by the density of collagen fibers correlated with high COMP levels in both cancer cells and stroma. This in turn could lead to exclusion of cytotoxic T-cells from accessing the cancer cells, a recognized immunotherapy resistance mechanism. Targeting COMP could therefore be considered as a novel therapeutic strategy in PB-type periampullary adenocarcinoma