Material and Methods Subject 173 myeloma patients 45

Material and Methods • Subject: 173 myeloma patients, 45 control • Plasma cells from the patients and control subjects • Oligonucleotide microarray profiling • Biochemical analysis(ELISA) • Immunohistochemistry analysis • Osteoblast differentiation assay

DDK 1: Wnt antagonist

Wnt signaling pathway • Wnt receptor: Frizzle family • Wnt coreceptor : lipoprotein receptor-related protein 5(LRP 5); DKK 1 bind to LRP 5 • Dishevelled protein pathway(G protein independent) • Inhibit beta-catenin phosphorylation and degradation • Beta-catenin accumulation and entry to nucleus • Beta-catenin activate Wnt gene

DKK 1 synthesis by Plasma Cells • 44/45 DKK 1 + myeloma: low grade morphologic findings, interstitial pattern • 19/20 DKK 1 - myeloma: high grade, plasmablastic morphologic findings, nodular pattern

DKK 1 expression and level

Effect of Bone Marrow Plasma on Osteoblast Differentiation in Vitro • Bone morphogenetic protein type 2 (BMP 2) • Alkaline phosphatase: marker for osteoblast • C 2 C 12: mesenchymal precursor cells • C 2 C 12 + BMP 2 + DKK 1 + anti-DKK 1

Role of DKK 1 in myeloma • In vitro study of mesenchymal stem cells exposed to different levels of DKK 1 • The role of DKK 1 in causing immunosuppression and anemia • Direct blocking Wnt signaling • Indirect inhibition of osteoblast differentiation

DKK 1 at different stages of myeloma • DKK 1 rarely detected in patients with monoclonal gammopathy of undermined significance or plasma cell leukemia • Stage specific expression • Down-regulation of DKK 1 by osteoclasts • RNAK ligands: abudnant in osteoblast precursor cells, not mature osteoblasts • RNAK ligands: activate osteoclasts