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Ferrata Storti Foundation
Haematologica 2021 Volume 106(9):2334-2344
Three-dimensional co-culture model of chronic lymphocytic leukemia bone marrow microenvironment predicts patient-specific response to mobilizing agents
Federica Barbaglio,1,2* Daniela Belloni,2* Lydia Scarfò,2,3,4
Francesca Vittoria Sbrana,1 Maurilio Ponzoni,3,5 Lucia Bongiovanni,5
Luca Pavesi,1 Desiree Zambroni,6 Kostas Stamatopoulos,7,8 Valeria R. Caiolfa,6,9 Elisabetta Ferrero,2 Paolo Ghia2,3,4 and Cristina Scielzo1
1Unit of Malignant B Cells Biology and 3D Modelling, Division of Experimental Oncology, IRCCS Ospedale San Raffaele, Milan, Italy; 2Unit of B Cell Neoplasia, Division of Experimental Oncology, IRCCS Ospedale San Raffaele, Milan, Italy;
3Università Vita-Salute San Raffaele, Milan, Italy; 4Strategic Research Program on CLL, Division of Experimental Oncology, IRCCS Ospedale San Raffaele, Milan, Italy; 5Pathology Unit, IRCCS Ospedale San Raffaele, Milan, Italy; 6Center for Experimental Imaging, IRCCS, Ospedale San Raffaele, Milan, Italy; 7Hematology Department and HCT Unit, G. Papanicolaou Hospital, Thessaloniki, Greece; 8Institute of Applied Biosciences, Centre for Research and Technology Hellas, Thessaloniki, Greece and 9Centro Nacional de investigaciones Cardiovasculares, (CNIC), Madrid, Spain
*FB and DB contributed equally as co-first authors.
ABSTRACT
Chronic lymphocytic leukemia (CLL) cells disseminate into sup- portive tissue microenvironments. To investigate the mechanisms involved in leukemic cell tissue retention we developed a three- dimensional bone marrow (BM) microenvironment that recreates the interactions between CLL and BM stromal cells inside a scaffold within a bioreactor. Our system allows the parallel analysis of CLL cells retained inside the scaffold and those released in the presence/absence of phar- macological agents, mimicking tissue and circulating cell compartments, respectively. CLL cells can be retained within the scaffold only in the presence of microenvironmental elements, which through direct contact downregulate the expression of HS1 cytoskeletal protein in CLL cells. Consistent with this, the expression of HS1 was lower in CLL cells obtained from patients’ BM than in CLL cells circulating in the peripheral blood. Moreover, we demonstrate that CLL cells with inactive HS1, impaired cytoskeletal activity and a more aggressive phenotype are more likely to be retained within the scaffold despite the presence of ibrutinib, whose mobilizing effect is mainly exerted on those with active HS1, ensuing dynamic cytoskeletal activity. This differential effect would not otherwise be assessable in a traditional two-dimensional system and may underlie a distinctive resistance of single CLL clones. Notably, CLL cells mobilized in the peripheral blood of patients during ibrutinib ther- apy exhibited activated HS1, underscoring that our model reliably mir- rors the in vivo situation. The three-dimensional model described herein is suitable for reproducing and identifying critical CLL-BM interactions, opening the way to pathophysiological studies and the evaluation of novel targeted therapies in an individualized manner.
Introduction
Chronic lymphocytic leukemia (CLL) is characterized by a progressive expansion of clonal CD5+ B lymphocytes that accumulate and traffic between the peripheral blood (PB), bone marrow (BM) and secondary lymphoid organs.1,2 In those sites, CLL cells are extremely dependent on and reactive to the microenvironment (i.e., stromal, endothelial cells and immune cells) and proliferate in so-called “proliferation centers”,
Correspondence:
CRISTINA SCIELZO
scielzo.cristina@hsr.it
Received: January 28, 2020. Accepted: July 17, 2020. Pre-published: July 30, 2020.
https://doi.org/10.3324/haematol.2020.248112 ©2021 Ferrata Storti Foundation
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haematologica | 2021; 106(9)
Chronic Lymphocytic Leukemia
ARTICLE