Surface molecule CD229 as a novel target for the diagnosis and treatment of multiple myeloma

Abstract

The online version of this article has a Supplementary Appendix. BackgroundTo date, multiple myeloma remains an incurable malignancy due to the persistence of minimal residual disease in the bone marrow. In this setting, monoclonal antibodies against myelomaspecific cell surface antigens represent a promising therapeutic approach, which is however hampered by a lack of appropriate target structures expressed across all pathogenic myeloma cell populations. We, therefore, investigated functionally relevant immunoreceptors specifically associated with myeloma cells as well as their clonogenic precursors. Design and MethodsPotential target proteins were identified using antibody arrays against phosphorylated immunoreceptors with lysates from myeloma cell lines. CD229 expression was confirmed in primary myeloma cells by reverse transcriptase polymerase chain reaction, western blot, fluorescence-activated cell sorting, and immunohistochemistry. Apoptosis, clonogenic growth, and sensitivity to chemotherapy were determined following short-interfering RNA-mediated downregulation of CD229. Antibody-dependent cellular and complement-dependent cytotoxicity were analyzed using a monoclonal antibody against CD229 to demonstrate the antigen's immunotherapeutic potential. ResultsOur screening assay identified CD229 as the most strongly over-expressed/phosphorylated immunoreceptor in myeloma cell lines. Over-expression was further demonstrated in the CD138-negative population, which has been suggested to represent myeloma precursors, as well as on primary tumor cells from myeloma patients. Accordingly, CD229 staining of patients' bone marrow samples enabled the identification of myeloma cells by flow cytometry and immunohistochemistry. Down-regulation of CD229 led to a decreased number of viable myeloma cells and clonal myeloma colonies, and enhanced the anti-tumor activity of conventional chemotherapeutics. Targeting CD229 with a monoclonal antibody resulted in complement-and cell-mediated lysis of myeloma cells. ConclusionsOur results demonstrate that the immunoreceptor CD229 is specifically over-expressed on myeloma cells including their clonogenic precursors and contributes to their malignant phenotype. Monoclonal antibodies against this protein may represent a promising diagnostic and immunotherapeutic instrument in this disease. Haematologica 2011;96(10):1512-1520. doi:10.3324/haematol.2010 Surface molecule CD229 as a novel target for the diagnosis and treatment of multiple myeloma

CD229 is expressed on the surface of plasma cells carrying an aberrant phenotype and chemotherapy-resistant precursor cells in Multiple Myeloma

“…11 Here, we have shown that CD229 is homogenously expressed on the malignant plasma cells across all plasma cell dyscrasias while it shows much lower levels of expression on the other leukocyte subpopulations present in the patients' bone marrow. We have also shown that CD229 is preferentially expressed on those bone marrow-infiltrating plasma cells showing an abnormal, more "malignant" phenotype as indicated, for example, by expression of CD56.…”

“…11 In particular, the unintended targeting of normal T and NK cells by CD229-specific CAR T cells may affect strategies for their use as a therapeutic option. However, recent data suggested that the targeting of T cells by cross-reactive CARs may not be as problematic as expected.…”

CD229 Expression on Bone Marrow Plasma Cells from Patients with Multiple Myeloma and Monoclonal Gammopathies of Uncertain Significance

“…Some interesting findings have shown that CD229 is constantly present on the surface of human bone marrow PCs, being expressed in myeloma cell lines, multiple myeloma (MM) and monoclonal gammopathies of uncertain significance (MGUS). Thus, CD229 has been proposed as a novel target for the diagnosis and treatment of MM [3]. …”

Targeting of Ly9 (CD229) Disrupts Marginal Zone and B1 B Cell Homeostasis and Antibody Responses

“…Ly9 is also expressed in virtually all chronic lymphocytic leukemias (CLL) (12). Its expression is not lost during B cell differentiation into plasma cells, and it is highly expressed on multiple myeloma cells (13). Ly9 has four extracellular Ig-like domains, and as has been shown with other SLAMF members, the N-terminal Ig domain of Ly9 mediates homophilic adhesion (14).…”