Abstract 4855: Proteomic characterization of the extracellular matrix in multiple myeloma
Authors: Glavey Siobhan V. , Naba Alexandra , Gambella Manuela , Rocci Alberto , Sacco Antonio , Asara John , Palumbo Antonio , Hynes Richard O. , Roccaro Aldo M. , Ghobrial Irene M. 
Published: May 2015

Background:

The extracellular matrix is a major component of the tumor microenvironment, contributing to the regulation of cell survival, proliferation, differentiation and metastasis. In multiple myeloma (MM) ECM components, such as integrins, fibronectin and collagens, have been shown are critical to the pathogenesis of MM and the development of drug resistance. To date, despite some knowledge of the composition of the ECM in tumors, detailed profiling of the composition of the ECM in MM has not been carried out. Recent advances in proteomics have led to the characterization of the ECM and ECM-related proteins (“matrisome”) in normal human tissues and tumors in a systematic and comprehensive approach.

Methods:

Tumor Xenograft models; MM1S-GFP-Luc+ cells (5x106) were injected intravenously into SCID-Bg mice (n=4/group) and animals underwent weekly bioluminescent imaging (BLI). Mice were sacrificed after two weeks in order to mimic early tumor development (luminescence= 1x105 p/sec/cm2/sr). and compared to mice demonstrating high tumor burden (1x108 p/sec/cm2/sr) 5 weeks post injection. Human bone marrow aspirates; Whole bone marrow was obtained from MM patients (newly diagnosed n=9, relapsed n=9) and healthy human donors (n=9) following written informed consent. Sequential extractions of whole pooled bone marrow from mice and individual bone marrow humans was performed using the CNMCS(Cytosol/Nucleus/Membrane/Cytoskeleton) Compartmental Protein Extraction Kit (Cytomol, CA). Following this, proteins underwent reversed-phase high performance liquid chromatography followed by tandem mass spectrometry (MS). Identified peptide spectra were counted as a semi quantitative measure of abundance.

Results:

We detected a total of 1202, 982 and 329 unique proteins from enriched whole bone marrow samples from relapsed patients, newly diagnosed patients and mice, respectively. Of these, critical ECM components such as laminins, matrix metalloproteinases and collagens were found to be enriched in human MM ECM in comparison to healthy donors with increased abundance apparent with disease progression in mice. Specifically Bone Marrow Proteoglycan and Proteoglycan 3 were amongst the ECM proteins significantly enriched in the ECM of newly diagnosed patients. PRG3 is a p53 responsive gene that has been demonstrated to be upregulated in apoptotic cells in several cancers, including CLL. The ECM protein Elastin, the expression of which is closely associated with the invasive/metastatic potential of various cancer types, was also upregulated in the MM ECM where it may play an important role in the tumor-ECM interaction.

Conclusions:

We have profiled the ECM in MM using mass spectrometry with a view to determining the specific components that may be important in MM disease biology. Through this approach molecular mechanisms that influence MM development and progression can be uncovered and potential targets for therapy identified.

Citation Format: Siobhan V. Glavey, Alexandra Naba, Manuela Gambella, Alberto Rocci, Antonio Sacco, John Asara, Antonio Palumbo, Richard O. Hynes, Aldo M. Roccaro, Irene M. Ghobrial. Proteomic characterization of the extracellular matrix in multiple myeloma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4855. doi:10.1158/1538-7445.AM2014-4855