Angiogenic switch during 5T2MM murine myeloma tumorigenesis: role of CD45 heterogeneity

The active role of angiogenesis during disease progression is well recognized in solid tumors. In hematologic malignancies such as multiple myeloma (MM), it is not known whether tumor neovascularization is an epiphenomenon or whether it is actively involved in disease progression. At clinical presentation, myeloma disease and the associated angiogenesis are both well established. Here the 5T2MM murine model was used to analyze anglogenesis during preclinical myeloma stages. Bone marrow (BM) of 5T2MM-inoculated mice was analyzed at weekly intervals until the end stage of the disease. Histologic analysis and assessment of microvessel density (MVD) by CD31 staining demonstrated a preangiogenic stage of small tumor aggregates followed by an anglogenic switch and subsequently an anglogenic stage of progressive tumor growth and large, confluent tumor nodules. Flow cytometric analysis that indicated an increase in percentage CD45(-) MM cells preceded the anglogenic switch. Real-time polymerase chain reaction (RT-PCR) of sorted CD45(+) and CD45(-) MM cells indicated higher vascular endothelial growth factor 120 (VEGF120) and VEGF164 transcripts in CD45(-) MM cells. VEGF enzyme-linked immunosorbent assay (ELISA) revealed high secretion by CD45(-) MM cells but no protein secretion by CD45(+) MM cells, indicating angiogenic heterogeneity among the MM cells. These data suggest that, like in solid tumors, angiogenic switch and anglogenic heterogeneity exist in MM.