An Italian survey on the use of denosumab for the management of skeletal-related events in patients with bone metastases.

OBJECTIVE: The Italian Association for Medical Oncology (AIOM) recommends preventive treatment of skeletal-related events in order to improve survival and the quality of life of patients with advanced malignancies. The aim of the study was to evaluate whether routine clinical practice is in agreement with recommendations about the use of denosumab. PATIENTS AND METHODS: A survey was carried out in Italy in the oncological setting. RESULTS: The answers to the survey showed that a large proportion of patients with metastases from solid tumors receive treatment; almost all oncologists administered denosumab every 4 weeks but for a shorter period of time than recommended. CONCLUSIONS: This survey showed that Italian oncologists favor the use of bone-targeted therapy to prevent skeletal-related events in patients affected by metastatic breast, prostate or lung cancer, in agreement with current recommendations.

ANG II potentiates mitogenic effect of norepinephrine in vascular muscle cells: role of FGF-2.

We examined the possible cooperation between norepinephrine (NE) and ANG II on proliferation of cultured vascular smooth muscle cells (VSMCs) and the involved cellular mechanisms. Nanomolar NE concentrations stimulated VSMC proliferation through a prazosin-sensitive effect. The pretreatment of cells with 100 nM ANG II for 24 h significantly potentiated the NE-induced VSMC proliferation; this potentiating effect of ANG II was blocked by losartan but was unaffected by the AT(2) receptor antagonist PD-123177. ANG II pretreatment also potentiated the increase in inositol phosphate turnover and upregulated the cell expression of fibroblast growth factor (FGF-2) induced by NE. Anti-FGF-2 neutralizing antibodies prevented the potentiating effect of ANG II on NE-induced cell growth. Both ANG II and NE stimulated extracellular signal-related kinase (ERK1) activation, but an ANG II potentiation of the effect of NE on ERK1 activity was not detectable. Moreover, ANG II significantly increased protein synthesis but did not potentiate the hypertrophic effect of NE. These findings demonstrate that ANG II and NE cooperate in promoting VSMC growth and that FGF-2 upregulation is involved in this effect.

Inhibitory effect of full-length human endostatin on in vitro angiogenesis.

Endostatin, a C-terminal product of collagen XVIII, is a very powerful angiogenesis inhibitor. In vivo experiments in mice indicate that endostatin dramatically reduces tumor mass without causing the onset of any resistance to the treatment. Recently, a 12-aa shorter human endostatin has been purified from plasma, but is ineffective in in vitro angiogenesis assays. Here we report that the full-length human recombinant endostatin has a potent inhibitory activity in in vitro angiogenesis assays. Two powerful angiogenic factors were used to stimulate endothelial cells: FGF-2 and VEGF-165. Endostatin prevented cell growth both in the basal condition and after stimulation with FGF-2 or VEGF-165. Migration of microvascular endothelial cells toward FGF-2 or VEGF-165 was impaired, both when cells were pretreated with the inhibitor and when endostatin was added together with the growth factors. Furthermore, experiments of inhibition of proliferation performed on nonmicroendothelial cells showed that endostatin was ineffective. This study indicates that human endostatin is a potent angiogenesis inhibitor and suggests its use in human anticancer therapy.

Induction of inflammatory angiogenesis by monocyte chemoattractant protein-1.

Almost any growth of tumors is to some extent associated with an inflammatory reaction which may be anti-tumorigenic by acting directly on tumor cells or protumorigenic cells presumably by inducing tumor-associated angiogenesis. In this study, we have analyzed the angiogenesis-inducing capacity of monocyte chemoattractant protein-1 (MCP-1), a key regulatory molecule of monocyte trafficking to sites of inflammation. MCP-1 was found to be potently angiogenic when implanted into rabbit cornea, exerting potency similar to the specific angiogenic vascular endothelial growth factor (VEGF)-A(121). MCP-1-induced angiogenesis in the cornea is associated with prominent recruitment of macrophages, whereas VEGF-A(121)-induced corneal angiogenesis is devoid of inflammatory cell recruitment. Based on these findings, we studied MCP-1 expression and macrophage recruitment in human invasive ductal mammary carcinomas in comparison with the physiological angiogenic processes in bovine ovarian corpus luteum. Macrophage recruitment was always associated with MCP-1 expression. High macrophage counts in mammary tumors corresponded with poor prognosis. In contrast, physiological ovarian angiogenesis was associated with only minimal inflammatory recruitment of macrophages. Our data show that MCP-1 is an indirect inflammation-associated inducer of angiogenesis and demonstrate distinct qualitative differences between tumor angiogenesis in human mammary tumors and physiological angiogenesis in the ovary.

Biological parameters for the choice of antiangiogenic therapy and efficacy monitoring.

Angiogenesis is a tightly controlled process which depends on the balance between stimulating and inhibiting factors. When this balance is disrupted, angiogenesis acquires a pathological meaning. The list of molecules able to induce angiogenesis is heterogeneous with respect to their chemical characteristics and biological properties. Quantitative measurement of tumor angiogenesis is necessary for the choice of therapeutic strategies and as an endpoint for antiangiogenic therapy. We are developing a quantitative RT-PCR with measures the expression of specific factors in real time. With the use of this rapid technique, measurement of the expression of the angiogenic factors and inhibitors is also possible in specimens as small as biopsies.

Endothelial cell migration is induced by soluble P-selectin.

We have assessed the role of soluble P-selectin in promoting endothelial cell migration. Human endothelial cells (HUVEC) and bovine microvascular endothelial cells (CVEC) were assessed for migration in the Neuroprobe 48-well microchemotaxis chamber. Soluble P-selectin promoted a dose-dependent (0.1-10 nM) migration of both cell types, with maximal response at 10 nM, producing approximately 60% increment over basal migration. Anti-P-selectin monoclonal antibody (5 microg/ml) selectively blocked P-selectin induced migration. Fibronectin and collagen were essential to disclose the migration induced by P-selectin. It is suggested that at vascular level in the presence of modifications of the extracellular matrix milieu, the production of soluble P-selectin could contribute to angiogenesis by promoting endothelial cell migration.

The prognostic significance of histologic regression in cutaneous melanoma.

A retrospective evaluation of 201 stage I cutaneous melanomas was performed to investigate the prognostic significance of histological regression (present in 67 cases). Thin melanomas showed regression more frequently than thick lesions (48% less than or equal to 0.75 mm vs 12% greater than 3 mm). The mean disease-free interval was 33.53 months in regressing tumours and 19.9 in non-regressing tumours (p = 0.07): differences between the survival curves were not significant (p = 0.61). Metastases developed in 13 (19.40%) patients with regressing tumours and by 40 (29.85%) patients with non-regressing tumours. Although we observed a higher frequency of regression in thin melanomas we could not demonstrate an influence of regression on disease-free interval and survival.

Melanocytic naevi: clinical features and correlation with the phenotype in healthy young males in Italy.

Six-hundred and fifteen young healthy Italian males were examined for the number, localization and clinical features of melanocytic naevi. The data were correlated with the phenotype and skin type of the subjects. Eighty-eight per cent had melanocytic naevi, 7% congenital naevi and 21.3% atypical naevi with a diameter greater than 5 mm and indistinct or irregular borders and were variable in colour. Of the sample only 2.43% had more than 30 naevi. Those subjects with blond or red hair, fair skin, blue or green eyes and with a skin type of I and II had higher numbers of naevi which were more often atypical than those subjects with dark hair, brown eyes and who were skin types III-V.

Phase II study of interferon alpha-2a and dacarbazine in advanced melanoma.

Based on the report of some activity of combination therapy with dacarbazine (DTIC) and interferon alpha-2a (rIFN alpha-2a) in disseminated melanoma, we conducted a phase II study to determine the feasibility and efficacy in a large series of patients. DTIC was administered in 79 patients at the dose of 800 mg/m2 every 3 weeks and rIFN alpha-2a was given daily at the dose of 9 X 10(6) IU for the first 10 weeks and three times a week thereafter. Among the 75 evaluable patients, 25% achieved an objective response, with 8% complete and 17% partial remissions. The regression occurred within a mean time of 1.9 +/- 1.03 months from starting therapy and the mean duration of response was 8.2 +/- 4.2 months. The major side effects were vomiting, anorexia, fever, fatigue, and myalgia. There was one death related to sepsis after myelosuppression. In the other patients bone marrow and liver toxicities were not remarkable. Our data reveal that a combination regimen of rIFN alpha-2a with a cytotoxic agent has some therapeutic activity in the management of advanced malignant melanoma.

Ki67 antigen expression correlates with tumor progression and HLA-DR antigen expression in melanocytic lesions.

Advanced steps of tumor progression are generally characterized by an increased growth fraction within the neoplastic cell population. The presence of a relevant growth fraction is also related to widely accepted prognostic parameters in some human malignancies. Our aims were to evaluate the presence of a growth fraction with Ki67 monoclonal antibody (MoAb), and to correlate it with tumor progression and HLA-DR antigen expression in 88 melanocytic lesions. The lesions were 19 acquired melanocytic nevi, 58 primary melanomas [divided into 26 superficial spreading melanomas (SSM), 24 superficial spreading melanomas with nodular areas (SS + NM), and five nodular melanomas (NM)], and 11 metastases from malignant melanomas. Ki67 MoAb stained 16%, 19%, 71%, 100%, and 82% of nevi, SSM, SS + NM, NM, and metastases, respectively. Among primary melanomas, Ki67 MoAb stained 12%, 28%, 50%, and 70% of tumors less than 0.75, 0.75-1.49, 1.5-2.9, and greater than or equal to 3 mm thick, respectively. A concordant reactivity pattern for Ki67 and HLA-DR antigens was found in 72% of lesions (p less than 0.0001). We have shown that a representative growth fraction (ie, Ki67 reactivity) is present in melanocytic lesions only in advanced steps of tumor progression and correlates with HLA-DR antigen expression. Despite the different biologic values of Ki67 and HLA-DR antigens, we suggest the joint evaluation of both antigens as a useful marker of aggressive behavior in melanoma.

HLA-DR and 96-K antigens and intratumoral lymphocytic infiltrate in primary cutaneous melanoma as markers of tumor progression.

It was recently demonstrated that HLA-DR antigens in primary melanomas are correlated with 96-K antigen expression and with the presence of an intratumoral lymphocytic infiltrate (ILI). As HLA-DR antigens are known to be associated with advanced phases of tumor progression in melanocytic lesions, we investigated whether primary melanomas in different progression phases could be distinguished on the basis of the expression of HLA-DR and 96-K antigens and of the presence of an ILI. A concordant presence of the two antigens and of an ILI was frequent both in vertical growth pattern melanomas and in melanomas thicker than 1 mm. On the contrary, both antigens and the ILI were frequently absent both in thin melanomas (less than 1 mm) and in radial growth pattern melanomas. These observations suggest that a joint study of HLA-DR and 96-K antigens and of the ILI is useful to define the phase of progression of primary melanomas.

Superficial spreading melanoma with a nodular area: antigenic phenotype of the radial and vertical growth areas.

We investigated antigenic features associated with different tumor progression steps in primary melanoma, interpreted as different patterns of growth (radial and vertical) in the same and in different lesions. Thirty-eight primary melanomas were examined: 18 superficial spreading malanomas, 13 superficial spreading melanomas with a nodular area and 7 nodular melanomas. 225.28,763.74, CL.203, VF19LL209, VF19LL217, Q5.13, W6.32 and anti-HLA-DR monoclonal antibodies were used. Phenotypic differences between radial and vertical growth areas were observed but no statistical significance could be found.

Multiple marker studies on a malignant fibrous histiocytoma with primary cutaneous localization.

Continuing controversy exists concerning a possible relation between neoplastic cells of malignant fibrous histiocytoma (MFH) and the mononuclear phagocyte system. The aim of this study was to investigate the membrane and cytoenzymatic phenotype of a primary cutaneous MFH, storiform pleomorphic type, and to compare these data with ultrastructural observations. Cytoplasmic proteins (acid phosphatase, non specific esterase, alpha-1 antitrypsin, and lysozyme) suggestive of a mononuclear phagocyte origin were demonstrated in varying amounts in neoplastic cells infiltrating the dermis. Consistent with these data, two (LeuM3 and OKM5) out of four (OKM1 and LeuM1) monoclonal antibodies directed against mononuclear phagocyte antigens stained most of the neoplastic cells. Class II MCH antigens (DR and DQ) were variably expressed on distinct groups of neoplastic cells, suggesting different activation/differentiation states. The results favor the view that the present case of primary cutaneous MFH was of mononuclear phagocyte origin. However, the observed phenotypic profile was expressed on neoplastic cells irrespective of their ultrastructural morphology (histiocytic or fibroblastic). Together with previous data in the literature, the latter finding corroborates the view that distinction between these two cell types in MFH is likely to reflect divergent growth and differentiation patterns rather than histogenesis.

Antigenic phenotype of radial growth phase melanomas with or without a vertical growth phase portion.

The expression of 4 melanoma-associated antigens and of class I and II HLA antigens was investigated in 12 superficial spreading melanomas (SSM) and in 8 SSM with a vertical growth pattern portion (SS + NM) by the use of monoclonal antibodies and an indirect immunoperoxidase procedure. Monoclonal antibodies 225.28, 763.74, CL.203, VF19-LL217, Q5-13, W6-32 and anti-HLA-DR, were used. Each antigen was more frequently expressed by SS + NM on the whole than by SSM and also by the radial growth pattern portions of SS + NM than by SSM. Vertical growth pattern portions of SS + NM were not antigenically similar to radial growth pattern portions in the same tumors. The high frequency of antigen expression in radial growth pattern melanomas seems to be associated with the appearance of a more invasive cell population.