Aberrant Levels of miRNAs in Bone Marrow Microenvironment and Peripheral Blood of Myeloma Patients and Disease Progression Weixin Wang, Meghan Corrigan-Cummins,

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Aberrant Levels of miRNAs in Bone Marrow Microenvironment and Peripheral Blood of Myeloma Patients and Disease Progression Weixin Wang, Meghan Corrigan-Cummins, Emily A. Barber, Layla M. Saleh, Adriana Zingone, Azam Ghafoor, Rene Costello, Yong Zhang, Roger J. Kurlander, Neha Korde, Aldo M. Roccaro, Irene M. Ghobrial, Ola Landgren, Katherine R. Calvo The Journal of Molecular Diagnostics Volume 17, Issue 6, Pages 669-678 (November 2015) DOI: 10.1016/j.jmoldx.2015.06.006 Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 1 Myeloma miRNA signature in the bone marrow extracellular microenvironment. Heatmap on the basis of hierarchical clustering of 111 miRNAs differentially expressed in the bone marrow extracellular microenvironment of MM in comparison with healthy control marrow as measured by miRNA array (P < 0.05). Twenty MM and eight control samples are shown in the heatmap. Ctrl, control; MM, multiple myeloma. The Journal of Molecular Diagnostics 2015 17, 669-678DOI: (10.1016/j.jmoldx.2015.06.006) Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 2 miRNAs differentially expressed in bone marrow microenvironment of myeloma. Scatter plots of selected miRNAs that were significantly decreased in the MM extracellular BM microenvironment in comparison with healthy control marrow as validated by qPCR. ∗∗∗P < 0.001. BM, bone marrow; MM, multiple myeloma; qPCR, quantitative real-time PCR. The Journal of Molecular Diagnostics 2015 17, 669-678DOI: (10.1016/j.jmoldx.2015.06.006) Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 3 miRNAs differentially expressed in the serum of myeloma. Scatter plots of selected miRNAs significantly differentially expressed in the serum of MM in comparison with serum of healthy controls as measured by qPCR. ∗∗P < 0.01, ∗∗∗P < 0.001. MM, multiple myeloma; qPCR, quantitative real-time PCR. The Journal of Molecular Diagnostics 2015 17, 669-678DOI: (10.1016/j.jmoldx.2015.06.006) Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 4 miRNAs differentially expressed in serum of MGUS, SMM, and myeloma. Scatter plots of selected miRNAs differentially detected in peripheral blood of MGUS, SMM, and (MM) by qPCR. In comparison with the serum of healthy controls, Let-7i, miR-15a, miR-16, and miR-106b are significantly decreased in serum of MGUS, SMM, and MM; Let-7a, let-7b, miR-15b, and miR-20a are significantly decreased in serum of SMM and MM; and miR-21, miR-223, and miR-361 are significantly decreased in the serum of MM. Values for MGUS and SMM were not statistically significant. ∗P < 0.05 compared to control. MGUS, monoclonal gammopathy of undetermined significance; MM, multiple myeloma; qPCR, quantitative real-time PCR; SMM, smoldering myeloma. The Journal of Molecular Diagnostics 2015 17, 669-678DOI: (10.1016/j.jmoldx.2015.06.006) Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 5 miRNAs differentially expressed in plasma cells of MGUS and myeloma. Scatter plots of selected miRNAs differentially expressed in CD138+ plasma cells of MGUS and MM. ∗∗P < 0.01. MGUS, monoclonal gammopathy of undetermined significance; MM, multiple myeloma. The Journal of Molecular Diagnostics 2015 17, 669-678DOI: (10.1016/j.jmoldx.2015.06.006) Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions