Microvessel density, endothelial-cell proliferation and carbonic anhydrase IX expression in haematological malignancies, bone-marrow metastases and monoclonal gammopathy of undetermined significance

Abstract

The aim of the study was to compare the angiogenic status, potential qualitative differences in microvessels and carbonic anhydrase IX expression in bone-marrow (BM) metastases and different haematological tumours at time of diagnosis. The microvessel density (MVD), endothelial-cell proliferation (ECP) and carbonic anhydrase IX (CA IX) immunoreactivity were determined on 210 trephine biopsies from 57 patients with multiple myeloma (MM), 13 with acute myeloid leukaemia (AML), 48 with chronic myeloproliferative syndrome (CMPS), 26 with chronic lymphocytic leukaemia (CLL), 25 with epithelial BM metastases, 18 with monoclonal gammopathy of undetermined significance (MGUS) and from a control group composed of 23 patients without haematological neoplasm. There was an increased MVD and ECP in epithelial BM metastases, MM, AML, CMPS and in a part of CLL. While an ECP greater than 0 was detected in 72% of MM, 75% of CMPS and 92% of AML, it was invariably observed (100%) in the BM metastases. The absence of ECP together with a MVD comparable with the control group in our MGUS cases supports the view that MGUS is a pre-angiogenic condition. Qualitative differences in microvessels were associated with growth patterns in MM and CLL and were observed between the different entities of CMPS. In one-third of the epithelial BM metastases, there was a focal CA IX immunoreactivity, which was never observed in the haematological diseases.

Part of the multiple myeloma-associated microvessels is functionally connected to the systemic circulation: a study in the murine 5T33MM model

“…One can hypothesize that the expansion of the MM population within the confinement of the bone could lead to an increased pressure exerted on the microvessel wall. The appearance of slit-like lumina of MM-associated microvessels in densely infiltrated areas, as we [8] and others already observed [16] in human BM specimens, is compatible with this hypothesis. The overexpression of vascular endothelial growth factor by human MM [7] and by 5T33MM cells [21] leads to angiogenesis with a partially disorganized vascular tree composed of tortuous microvessels.…”

Antitumour and antiangiogenic effects of Aplidin® in the 5TMM syngeneic models of multiple myeloma

“…Sections were stained for the presence of CD31 (PECAM-1) to identify the presence of microvessels or for Ki-67 to identify the proliferative cells (De Raeve et al, 2004). For CD31 retrieval, sections were incubated in trypsin to promote antigen retrieval and blocked with normal goat serum for 30 min.…”

Choice of endothelial marker is crucial for assessment of bone marrow microvessel density in chronic lymphocytic leukemia

“…Assessment of bone marrow microvessel density has been widely reported as a standard method of investigation of angiogenesis in haematological malignancies (11–14). Nevertheless, results of studies on marrow neovascularization in CLL are inconsistent (15–20), possibly due to the limited number of patient cohorts and different methods of MVD assessment methodology, including use of different monoclonal antibodies for endothelial cell staining. In the present study, we examined bone marrow microvessel density in chronic lymphocytic leukemia patients using the two most commonly applied endothelial markers, CD34 and vWF.…”

“…It is well‐known that CLL cells are capable of producing many angiogenic molecules (4) and that various circulating angiogenic factors are elevated in peripheral blood of CLL patients (5–10). Elevated bone marrow neovascularization has been widely reported in various hematological malignancies (11–14); however, it is still controversial whether in CLL bone marrow is elevated, in part because methods of angiogenesis assessment are not directly comparable between published studies (due to different antibodies used for immunohistochemical identification of vessels and different methods of calculating microvessel density [MVD]) (15–20). Moreover, there is insufficient data on the association of bone marrow angiogenesis with prognostic factors, especially modern ones (IgVH mutation status, genetic abnormalities).…”