Page 163 - Haematologica-April 2018

Plasma Cell Disorders
Modeling multiple myeloma-bone marrow inter- actions and response to drugs in a 3D surrogate microenvironment
DanielaBelloni,1 SilviaHeltai,1 MaurilioPonzoni,2,3 AntonelloVilla,4 Barbara Vergani,4 Lorenza Pecciarini,2 Magda Marcatti,5 Stefania Girlanda,5 Giovanni Tonon,6 Fabio Ciceri,3,5 Federico Caligaris-Cappio,1,3,7
Marina Ferrarini1* and Elisabetta Ferrero1*
1Division of Experimental Oncology, IRCCS San Raffaele Scientific Institute; 2Pathology Unit, IRCCS San Raffaele Scientific Institute; 3Vita-Salute San Raffaele University; 4Consorzio MIA, University of Milano-Bicocca; 5Hematology, IRCCS San Raffaele Scientific Institute; 6Functional Genomics of Cancer Unit, Division of Experimental Oncology, San Raffaele Scientific Institute and 7AIRC, Milan, Italy
Ferrata Storti Foundation
Haematologica 2017 Volume 103(4):707-716
*MF and EF contributed equally to this work
ABSTRACT
Multiple myeloma develops primarily inside the bone marrow microenvironment, that confers pro-survival signals and drug resistance. 3D cultures that reproduce multiple myeloma-bone marrow interactions are needed to fully investigate multiple myeloma pathogenesis and response to drugs. To this purpose, we exploited the 3D Rotary Cell Culture System bioreactor technology for myeloma- bone marrow co-cultures in gelatin scaffolds. The model was validated with myeloma cell lines that, as assessed by histochemical and electron- microscopic analyses, engaged contacts with stromal cells and endothe- lial cells. Consistently, pro-survival signaling and also cell adhesion- mediated drug resistance were significantly higher in 3D than in 2D par- allel co-cultures. The contribution of the VLA-4/VCAM1 pathway to resistance to bortezomib was modeled by the use of VCAM1 transfec- tants. Soluble factor-mediated drug resistance could be also demonstrat- ed in both 2D and 3D co-cultures. The system was then successfully applied to co-cultures of primary myeloma cells-primary myeloma bone marrow stromal cells from patients and endothelial cells, allowing the development of functional myeloma-stroma interactions and MM cell long-term survival. Significantly, genomic analysis performed in a high- risk myeloma patient demonstrated that culture in bioreactor paralleled the expansion of the clone that ultimately dominated in vivo. Finally, the impact of bortezomib on myeloma cells and on specialized functions of the microenvironment could be evaluated. Our findings indicate that 3D dynamic culture of reconstructed human multiple myeloma microenvi- ronments in bioreactor may represent a useful platform for drug testing and for studying tumor-stroma molecular interactions.
Introduction
Tumors develop and progress within co-evolving microenvironments that affect both the fate of the tumor and its drug sensitivity. Response to drugs may be over- estimated on 2-dimensional (2D) cultures, and the discrepancy between pre-clinical findings and clinical outcomes can also be attributed to the failure of conventional 2D models.1-4 3D culture models closely reproduce tumor within its microenviron- ment, recapitulating tumor-stroma interactions and signaling.1-4 Multiple myeloma (MM) is a paradigm of tumor-stroma inter-dependence, as it develops almost exclu- sively within the bone marrow (BM),5-8 where MM cells establish tight contacts with the stroma, that in turn delivers pro-survival, anti-apoptotic signals and con- fers drug resistance.9 Accordingly, new drugs, including proteasome inhibitors, have been developed to target both MM cells and their BM microenvironment. However, the disease remains incurable predominantly due to development of drug resistance. Along this line, 3D models of MM cells inside their microenviron-
Correspondence:
ferrero.elisabetta@hsr.it or ferrarini.marina@hsr.it
Received: February 23, 2017. Accepted: December 27, 2017. Pre-published: January 11, 2018.
doi:10.3324/haematol.2017.167486
Check the online version for the most updated information on this article, online supplements, and information on authorship & disclosures: www.haematologica.org/content/103/4/707
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