Circulating osteopontin: a dual marker of bone destruction and angiogenesis in patients with multiple myeloma

The matrix protein osteopontin has been shown to be a marker of osteoclastic activity in multiple myeloma patients, as well as a regulator of angiogenesis. We measured serum levels of osteopontin in 50 untreated multiple myeloma patients (in 25, also after treatment) and examined the relation to markers of osteolytic and angiogenic activity. The median (range) of serum osteopontin was 85 (5-232) in the patient group vs. 36 (2-190) ng/ml in the control group. Serum osteopontin levels were significantly higher in patients with advanced stage or grade of myeloma disease. All patients with serum osteopontin levels >100 ng/ml had advanced stage (II or III) or high grade bone disease, whereas stage I or low grade patients had serum osteopontin levels <100ng/ml. Serum osteopontin levels significantly decreased after treatment. There was a positive correlation of osteopontin with the bone turnover marker N-terminal propeptide of procollagen type I (NTx) and the angiogenic markers vascular endothelial growth factor (VEGF) and bone marrow microvessel density (r: 0.35, 0.47 and 0.30 respectively, p < 0.05). These results support osteopontin as a dual marker of bone destruction and angiogenic activity in myeloma patients. Osteopontin represents a useful biomarker for monitoring myeloma disease activity

Investigation of the role of mast cells and CXC ELR motif (ELR+) chemokines in the angiogenesis process of multiple myeloma

Background: Multiple myeloma is a malignant plasma cell diseaes, being located in bonemarrow. Malignant cells interact with bone marrow microenvironment, modifying it.Angiogenesis represents the formation of new blood vessels from pre-existing ones, isdependent on those interactions, and finally is implicated in the development of tumorby multiple manners. Angiogenesis process has been studied in both benign andmalignant diseases included multiple myeloma, achieving the development of antiangiogenictherapies, even in myeloma, with satisfying results. Despite the progress in theunderstanding of myeloma pathophysiology, the disease remains incurable. Therefore,the investigation of implicated mechanisms in the biology of myeloma evolution isnessecary.Aim of the study: The aim of the study was to investigate the role of ELR CXC+chemokines and mast cells in angiogenic activity of multiple myeloma. Circulating levelsof those chemokines in the peripheral blood, as well mast cells’ location in bone marrowwere studied. Furthermore, their relationship with bone marrow angiogenic process wasalso studied.Patients and methods: Sixty-three myeloma patients were studied during diagnosis.Moreover, 30 of them achieving remission after therapy as well 30 healthy controls werealso studied. This was a study in 2 phases. Initially, ELR+ CXC chemokines (GRO-α,ENA-78 and ΙL-8) were measured in peripheral blood, along with the estimation of bonemarrow angiogenic activity, both directly, estimating bone marrow microvascular density(MVD) and indirectly, measuring circulating levels of known angiogenic cytokines, suchas HGF, VEGF, TNF-α, angiogenin and PDGF-AB. In the next phase, bone marrowmast cell density (MCD) was estimated, along with the estimation of bone marrowangiogenic activity, both directly, estimating bone marrow microvascular density (MVD) and indirectly, measuring circulating levels of known angiogenic cytokines, such asVEGF, bFGF, TNF-α, as well ELR+ CXC chemokines.Results: All measured variables were significantly increased in multiple myeloma patientscompared to healthy population. Furthermore, they were all increasing in parallel withdisease activity, with the exception of ENA-78. Similarly, all variables decreased aftereffective treatmen, with the exceptions of ENA-78 and GRO-α, where no difference wasfound. Significant positive correlations were found between GRO-α with HGF, VEGF,TNF-α, MVD and MCD. Significant positive correlations were also noted betweenΕΝΑ-78 with VEGF, TNF-α and MCD but not with MVD. IL-8 also correlated with TNF-α, PDGF-AB, angiogenin and MCD, but also not with MVD. Finally, MCDcorrelated with MVD and all angiogenic cytokines as well.Conclusions: Circulating levels of ELR+ CXC chemokines (IL-8, GRO-α and ΕΝΑ-78), as well bone marrow MCD are elevated in multiple myeloma patients, are dependenton disease stage and decrease after effective therapy. Their correlations with theangiogenic process suggest their participation in multiple myeloma angoiogenesis.Υπάρχοντα δεδομένα: Το πολλαπλό μυέλωμα είναι μια νεοπλασματική νόσος τωνπλασματοκυττάρων, εντοπιζόμενη στο μυελό των οστών. Αυτά τα νεοπλασματικά κύτταρααλληλοεπιδρούν με το μυελικό μικροπεριβάλλον, με αποτέλεσμα να το τροποποιήσουν. Ηαγγειογένεση, δηλαδή η δημιουργία νέων αγγείων από τα προϋπάρχοντα, εξαρτάται απόαυτές τις αλληλεπιδράσεις, και τελικά εμπλέκεται στην ανάπτυξη του όγκου με πολλαπλούςτρόπους. Η διαδικασία της αγγειογένεσης έχει ήδη μελετηθεί σε διάφορα καλοήθη καικακοήθη νοσήματα, συμπεριλαμβανομένου του μυελώματος, και ήδη έχουν προκύψειειδικές αντιαγγειογενετικές θεραπευτικές παρεμβάσεις, ακόμα και στο πολλαπλό μυέλωμαμε ικανοποιητική αποτελεσματικότητα. Όμως, παρά την εξέλιξη στην κατανόηση τηςπαθοφυσιολογίας της νόσου και στην κλινική εφαρμογή της, το πολλαπλό μυέλωμαπαραμένει ανίατο και συνεπώς απαιτεί περισσότερη διερεύνηση των εμπλεκόμενωνμηχανισμών στη βιολογία της εξέλιξής του.Σκοπός της μελέτης: Σκοπός της παρούσας διατριβής ήταν να μελετήσει το ρόλο τωνELR CXC+ χυμοκινών και των μαστοκυττάρων στην αγγειογενετική δραστηριότητα τουπολλαπλού μυελώματος. Μετρήθηκε η περιφερική ανίχνευση αυτών των χυμοκινών καθώςκαι η εντόπιση των μαστοκυττάρων στο μυελό και εξετάσθηκε η σχέση τους, τόσο με τηναγγειογενετική δραστηριότητα του μυελού.Υλικά και μέθοδοι: Μελετήθηκαν 63 ασθενείς κατά τη διάγνωση του μυελώματος, καθώςκαι 30 από αυτούς, που μπήκαν σε ύφεση μετά από θεραπεία. Επίσης μελετήθηκαν 30 υγιείςμάρτυρες. Η μελέτη έγινε σε 2 φάσεις. Αρχικά μελετήθηκαν οι ELR+ CXC χυμοκίνες(GRO-α, ENA-78 και ΙL-8) στο περιφερικό αίμα, σε συνδυασμό με την εκτίμηση τηςαγγειογενετικής δραστηριότητας του μυελού, μέσω άμεσης μέτρησης της μικροαγγειακήςπυκνότητας του (MVD) και μέσω έμμεσης μέτρησης των αγγειογενετικών κυτταροκινώνHGF, VEGF, TNF-α, αγγειογενίνη και PDGF-AB, στο περιφερικό αίμα.. Στην επόμενηφάση μετρήθηκε η μαστοκυτταρική πυκνότητα στο μυελό (MCD) σε συνδυασμό με τηνεκτίμηση της αγγειογενετικής δραστηριότητάς του, μέσω άμεσης μέτρησης της MVD τουκαι μέσω έμμεσης μέτρησης των αγγειογενετικών κυτταροκινών VEGF, bFGF, TNF-ακαθώς των ELR+ CXC χυμοκινών.Αποτελέσματα: Τα επίπεδα των ELR+ CXC χυμοκινών, των μετρούμενων αγγειογενετικώνπαραγόντων καθώς και η MVD και η MCD στο μυελό, ήταν όλα στατιστικά σημαντικάαυξημένα στους ασθενείς σε σχέση με τους υγιείς μάρτυρες. Επίσης όλες οι παραπάνωπαράμετροι αυξάνονταν με την πρόοδο του σταδίου της νόσου, με εξαίρεση την ΕΝΑ-78.Παρόμοια, όλες οι παραπάνω παράμετροι ελαττώνονταν στατιστικά σημαντικά μετά από αποτελεσματική θεραπεία, με εξαίρεση την ENA-78 και το GRO-α, όπου δε διαπιστώθηκεδιαφορά. Σημαντικές θετικές συσχετίσεις βρέθηκαν ανάμεσα στις τιμές του GRO-α με αυτέςτων HGF, VEGF, TNF-α αλλά και με τη MVD και MCD. Επίσης η ΕΝΑ-78συσχετίσθηκε με τον VEGF και τον TNF-α, καθώς και με τη MCD, αλλά όχι με τη MVD.H IL-8 σχετίσθηκε με τον TNF-α, τον PDGF-AB, την αγγειογενίνη και τη MCD, αλλά όχιμε τη MVD. Τέλος η MCD συσχετίσθηκε τόσο με τη MVD, όσο και με τις υπόλοιπεςαγγειογενετικές κυτταροκίνες..Συμπεράσματα: Τα κυκλοφορούντα επίπεδα των ELR+ CXC χυμοκινών (IL-8, GRO-ακαι ΕΝΑ-78) καθώς και η μαστοκυτατρική πυκνότητα του μυελού είναι αυξημένα στουςασθενείς με πολλαπλό μυέλωμα, εξαρτώνται από το στάδιο της νόσου και ελαττώνονται μετάαπό αποτελεσματική θεραπεία. Οι συσχετίσεις τους με την αγγειογενετική διαδικασίαπροτείνει τη συμμετοχή των ELR+ CXC χυμοκινών και των μαστοκυττάρων στηδιαδικασία της μυελωματικής αγγειογένεσης

Serum interleukin-17 and its relationship to angiogenic factors in multiple myeloma

Background: Interleukin-17 (IL-17) is a CD4 T-cell-derived mediator of angiogenesis that stimulates vascular endothelial cell migration and regulates the production of a variety of proangiogenic factors, such as tumor necrosis factor-α (TNF-α) and vascular endothelial cell growth factor (VEGF). Angiogenesis is implicated in the progression of multiple myeloma (MM). Methods: We measured serum levels of IL-17, TNF-α, and VEGF, as well as microvessel density (MVD) in 40 untreated MM patients. Results: Levels of IL-17 in the sera of patients with MM were higher than those in matched controls; however, the difference did not reach statistical significance. Serum levels of both TNF-α and VEGF in MM patients were significantly higher than those in controls (

Relationship between serum levels of vascular endothelial growth factor, hepatocyte growth factor and matrix metalloproteinase-9 with biochemical markers of bone disease in multiple myeloma

Background: Multiple myeloma is characterized by accumulation of plasma cells in the bone marrow, with osteolysis and increased marrow angiogenesis. We studied molecules involved in angiogenesis (MMP-9, HGF, VEGF) in relation to disease stage, extent of bone destruction, and markers of bone turnover (Ntx and PICP). Methods: MMP-9, HGF, VEGF were measured in the serum of 42 newly diagnosed myeloma patients and 24 controls with commercial ELISA kits. Urinary levels of Ntx were measured by ELISA, and serum PICP with RIA. Extent of radiological bone disease was graded into low and high score. Stage was estimated according to the Durie-Salmon criteria. Results: HGF, VEGF and Ntx were higher in patients than controls ( pb0.001). MMP-9 and PICP did not differ between patients and controls. HGF, VEGF, MMP-9 and Ntx increased significantly with disease stage (I to III, pb0.001) and PICP decreased significantly with advancing stage ( pb0.05). There was a positive correlation between HGF and MMP-9 (r: 0.36, pb0.01), VEGF and MMP-9 (r: 0.38, pb0.01), Ntx and MMP-9 (r: 0.39, pb0.01) and an inverse correlation between PICP and MMP-9 (r: −0.66, pb0.0001). Conclusions: Angiogenesis and bone destruction are closely interrelated in myeloma, and cytokine levels (MMP-9, VEGF and HGF) may be useful in monitoring progression

Serum levels of Interleukin-15 and Interleukin-10 and their correlation with proliferating cell nuclear antigen in multiple myeloma

In order to determine prognostic factors characterizing multiple myeloma (MM) cell kinetics, bone marrow proliferative activity and serum Interleukin-10 (IL-10), and Interleukin-15 (IL-15) levels were measured in 40 newly diagnosed MM patients, compared with 10- age and sex-matched-healthy controls. Cell proliferation was evaluated by employing a monoclonal antibody directed against the proliferating cell nuclear antigen (PCNA), whereas IL-10 and IL-15 were measured with quantitative sandwich enzyme immunoassay methods. IL-15, IL-10 and PCNA were higher in the patient group than in controls (P \u3c 0.001). IL-10 levels, and PCNA increased significantly with increasing Durie-Salmon disease stage (I-III, P \u3c 0.002, and P = 0.001, respectively). Serum IL-15 levels in MM stage III patients were elevated in comparison with stages I and II, the difference however, did not reach statistical significance. There was a significant positive correlation between serum IL-15 and IL-10 levels (r: 0.372, P \u3c 0.01), and between serum IL-10 and PCNA (r: 0.608, P \u3c 0.0001), as well as a positive correlation of serum IL-15 with PCNA, which marginally failed to reach statistical significance. Serum IL-15 levels are elevated in MM patients, increase with advancing stage, and correlate with Il-10 and PCNA. These proliferative factors may be useful in assessing disease progression in MM

Serotype distribution of Streptococcus pneumoniae in north-western Greece and implications for a vaccination programme

Serotypes and antibiotic sensitivities were determined for 338 strains of Streptococcus pneumoniae from children of north-western Greece with invasive pneumococcal disease (IPD), acute otitis media (AOM) and nasopharyngeal carriage. The most common serotypes among the isolates from IPD were 14 and 19F, while 3, 19F, 9V and 14 were the major cause of AOM. In these groups, the heptavalent conjugate vaccine for pneumococci (7vPCV) seems to cover 90.5% of the serotypes isolated from children less than 2 years old. Serotypes 23F and 6B were the most prevalent in carrier strains. Overall, 23.7% of the isolates were penicillin nonsusceptible (PNS), 97% were fully susceptible to cefotaxime, 29% were resistant to erythromycin, 11.2% to co-trimoxazole and 1.2% to clindamycin

Serum interleukin-17 and its relationship to angiogenic factors in multiple myeloma

Background: Interleukin-17 (IL-17) is a CD4 T-cell-derived mediator of angiogenesis that stimulates vascular endothelial cell migration and regulates the production of a variety of proangiogenic factors, such as tumor necrosis factor-alpha (TNF-alpha) and vascular endothelial cell growth factor (VEGF). Angiogenesis is implicated in the progression of multiple myeloma (MM). Methods: We measured serum levels of IL-17, TNF-alpha, and VEGF, as well as microvessel density (MVD) in 40 untreated MM patients. Results: Levels of IL-17 in the sera of patients with MM were higher than those in matched controls; however, the difference did not reach statistical significance. Serum levels of both TNF-alpha and VEGF in MM patients were significantly higher than those in controls (P&lt;0.001 in both instances). Levels of IL-17 in MM patients, both stage 11 and stage 111, were significantly higher than those of stage I patients (p=0.001 and p&lt;0.001, respectively). Similarly, higher values of VEGF (p&lt;0.001), TNF-a (p&lt;0.001), and MVD (p&lt;0.035) were associated with advanced disease stage. Serum values of IL-17 in MM patients correlated positively not only with VEGF (Spearman’s rho=0.606) and TNF-alpha (r=0.552; p&lt;0.001 in both instances), but also with MVD (r=0.385, p=0.014). In addition, a positive correlation was found between serum values of VEGF and TNF-alpha (r=0.657, p&lt;0.001), MVD and VEGF (r=0.353, p=0.026), and between MVD and TNF-alpha (r=0.506,p=0.001) in MM patients. Conclusion: These results suggest that IL-17 plays a role in the promotion of angiogenesis and associated disease progression in MM. (C) 2006 European Federation of Internal Medicine. Published by Elsevier B.V All rights reserved

Positive correlation between bone marrow mast cell density and ISS prognostic index in patients with multiple myeloma

We evaluated mast cell density (MCD) in myeloma bone marrow biopsies and correlated it with stage of disease and markers of angiogenesis. Fifty-three untreated myeloma patients and 28 of them responded to therapy were studied. Mast cells were highlighted using immunohistochemical stain for tryptase. Angiogenesis was evaluated measuring microvascular density and serum levels of basic-fibroblast growth factor and tumor necrosis factor-alpha. MCD was higher in untreated patients, compared to healthy population and responders. Significant association was found between MCD with angiogenesis and clinical stage of disease, suggesting that mast cells could be used as target for myeloma treatment. (C) 2013 Elsevier Ltd. All rights reserved

Increased serum levels of MIP-1alpha correlate with bone disease and angiogenic cytokines in patients with multiple myeloma

Many cytokines possess variable roles in the pathogenesis of multiple myeloma. Macrophage inflammatory protein-1alpha (MIP-1alpha) is an osteoclast-activating factor with a major role in myeloma bone disease. The aim of the study was to examine its participation in the angiogenic process of the disease. We measured, by enzyme-linked immunosorbent assays, its serum levels in 56 newly diagnosed myeloma patients, in several skeletal grades and stages of the disease and in 25 healthy controls. Concurrently, we measured serum levels of the angiogenic cytokines basic-fibroblast growth factor, hepatocyte growth factor and interleukin-18. All the above cytokines were higher in myeloma patients (p &lt; 0.001 for all cases) and were increasing in parallel with disease stage (p &lt; 0.001 for all cases) and skeletal grade (p &lt; 0.04 for MIP-1alpha and p &lt; 0.001 for the other cases). Moreover, positive correlations between MIP-1alpha and all the angiogenic cytokines were noted (p &lt; 0.001 for all cases). MIP-1alpha seems to be a predominant factor responsible for the enhancement of bone resorption and increased angiogenesis. The positive correlation between MIP-1alpha and the angiogenic chemoattractants supports the involvement of these factors in the biology of myeloma cell growth. Moreover, they could be used as possible therapeutic targets as well as markers of disease activity