Gene Expression Profiling of Bone Marrow Endothelial Cells in Patients with Multiple Myeloma

Roberto Ria,Angelo Vacca,Antonino Neri,Attilio Guarini,Domenico Ribatti,Michela Mattioli,Addolorata Maria Luce Coluccia,Annunziata De Luisi,Franco Dammacco,Domenica Ronchetti,Simona Berardi,Katia Todoerti,Fortunato Morabito,Maria Teresa Petrucci

doi:10.1158/1078-0432.ccr-09-0040

Abstract

To determine a "gene/molecular fingerprint" of multiple myeloma endothelial cells and identify vascular mechanisms governing the malignant progression from quiescent monoclonal gammopathy of undetermined significance. Comparative gene expression profiling of multiple myeloma endothelial cells and monoclonal gammopathy of undetermined significance endothelial cells with the Affymetrix U133A Arrays was carried out in patients at diagnosis; expression and function of selective vascular markers was validated by real-time reverse transcriptase-PCR, Western blot, and small interfering RNA analyses. Twenty-two genes were found differentially expressed (14 down-regulated and eight up-regulated) at relatively high stringency in multiple myeloma endothelial cells compared with monoclonal gammopathy of undetermined significance endothelial cells. Functional annotation revealed a role of these genes in the regulation of extracellular matrix formation and bone remodeling, cell adhesion, chemotaxis, angiogenesis, resistance to apoptosis, and cell-cycle regulation. Validation was focused on six genes (DIRAS3, SERPINF1, SRPX, BNIP3, IER3, and SEPW1) not previously found to be functionally correlated to the overangiogenic phenotype of multiple myeloma endothelial cells in active disease. The small interfering RNA knockdown of BNIP3, IER3, and SEPW1 genes affected critical multiple myeloma endothelial cell functions correlated with the overangiogenic phenotype. The distinct endothelial cell gene expression profiles and vascular phenotypes detected in this study may influence remodeling of the bone marrow microenvironment in patients with active multiple myeloma. A better understanding of the linkage between plasma cells and endothelial cells in multiple myeloma could contribute to the molecular classification of the disease and thus pinpoint selective gene targets for more effective antiangiogenic treatments.

Highlights

To determine a “gene/molecular fingerprint” of multiple myeloma endothelial cells and identify vascular mechanisms governing the malignant progression from quiescent monoclonal gammopathy of undetermined significance
The 663 probe sets found to be highly variable along the entire data set generated a dendrogram (Fig. 1A) with two major branches: one containing multiple myeloma endothelial cells and monoclonal gammopathy of undetermined significance endothelial cells, and the other grouping the Human umbilical vascular endothelial cells (HUVEC)
Previous gene expression profiling studies designed to identify genes perhaps involved in the initiation and progression of multiple myeloma [10, 18] revealed that monoclonal gammopathy of undetermined significance and multiple myeloma plasma cells can be distinguished from normal plasma cells, whereas their own differentiation is problematical. These findings suggest that modulation of the bone marrow microenvironment rather than genetic alterations of the tumor cells may partly account for the malignant conversion of monoclonal gammopathy of undetermined significance

Summary

Introduction

To determine a “gene/molecular fingerprint” of multiple myeloma endothelial cells and identify vascular mechanisms governing the malignant progression from quiescent monoclonal gammopathy of undetermined significance. Validation was focused on DIRAS3, SERPINF1, SRPX, BNIP3, IER3, and SEPW1 genes, which were not previously found to be functionally correlated to the overangiogenic phenotype of multiple myeloma endothelial cells.

Results

Conclusion