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Ferrata Storti Foundation
Haematologica 2021 Volume 106(10):2598-2612
Chronic lymphocytic leukemia cells impair osteoblastogenesis and promote osteoclasto- genesis: role of TNFα, IL-6 and IL-11 cytokines
Paolo Giannoni,1 Cecilia Marini,2 Giovanna Cutrona,3 Serena Matis,3 Maria Cristina Capra,3 Francesca Puglisi,1 Paola Luzzi,1 Simona Pigozzi,4 Gabriele Gaggero,4 Antonino Neri,5,6 Katia Todoerti,6 Fortunato Morabito,7,8 Adalberto Ibatici,9 Maurizio Miglino,9 Micaela Bergamaschi,9 Silvia Bruno,10 Gian Mario Sambuceti,11 Jean Louis Ravetti,4 Manlio Ferrarini,10 Franco Fais3,10 and
3 Daniela de Totero
1Department of Experimental Medicine, Biology Section, University of Genoa, Italy; 2CNR institute of Bioimages and Molecular Physiology, Milan, Italy; 3Molecular Pathology Unit, IRCCS-Ospedale Policlinico San Martino, Genoa, Italy; 4Pathological Anatomy Unit, IRCCS-Ospedale Policlinico San Martino, Genoa,Italy; 5Department of Oncology and Hemato-Oncology, University of Milan, Italy; 6Hematology Unit Fondazione IRCCS Ca’ Granda, Ospedale Maggiore Policlinico, Milan, Italy; 7Biotechnology Research Unit, Azienda Ospedaliera di Cosenza, Aprigliano, Cosenza, Italy; 8Department of Hematology and Bone Marrow Transplant, Augusta Victoria Hospital, Jerusalem, Israel; 9Hematology Clinic, IRCCS-Ospedale Policlinico San Martino, Genoa, Italy; 10Department of Experimental Medicine, Anatomy Section, University of Genoa, Genoa, Italy and 11Nuclear Medicine Unit, IRCCS-Ospedale Policlinico San Martino, Genoa, Italy
ABSTRACT
Bone skeletal alterations are no longer considered a rare event in chron- ic lymphocytic leukemia (CLL), especially at more advanced stages of the disease. This study is aimed at elucidating the mechanisms under- lying this phenomenon. Bone marrow stromal cells, induced to differenti- ate toward osteoblasts in osteogenic medium, appeared unable to complete their maturation upon co-culture with CLL cells, CLL-cell-derived condi- tioned media (CLL-cm) or CLL-sera (CLL-sr). Inhibition of osteoblast differ- entiation was documented by decreased levels of RUNX2 and osteocalcin mRNA expression, by increased osteopontin and DKK-1 mRNA levels, and by a marked reduction of mineralized matrix deposition. The addition of neutralizing TNFα, IL-11 or anti-IL-6R monoclonal antibodies to these co- cultures resulted in restoration of bone mineralization, indicating the involvement of these cytokines. These findings were further supported by silencing TNFα, IL-11 and IL-6 in leukemic cells. We also demonstrated that the addition of CLL-cm to monocytes, previously stimulated with MCSF and RANKL, significantly amplified the formation of large, mature osteo- clasts as well as their bone resorption activity. Moreover, enhanced osteo- clastogenesis, induced by CLL-cm, was significantly reduced by treating cultures with the anti-TNFα monoclonal antibody infliximab. An analo- gous effect was observed with the use of the BTK inhibitor, ibrutinib. Interestingly, CLL cells co-cultured with mature osteoclasts were protected from apoptosis and upregulated Ki-67. These experimental results parallel the direct correlation between amounts of TNFα in CLL-sr and the degree of compact bone erosion that we previously described, further strengthen- ing the indication of a reciprocal influence between leukemic cell expansion and bone structure derangement.
Introduction
In patients with chronic lymphocytic leukemia (CLL) skeletal erosion can be demonstrated by computed tomography scans of long bone shafts in subjects with more advanced disease (Binet C vs. Binet A).1,2 Moreover a few cases of CLL/small- cell lymphocytic lymphoma show osteolysis along with hypercalcemia.3-7 Axial bone structure alterations appear more frequently in CLL patients than in age-
Chronic Lymphocytic Leukemia
Correspondence:
DANIELA DE TOTERO
daniela.detotero@hsanmartino.it
Received: July 5, 2019. Accepted: August 21, 2020. Pre-published: August 27, 2020.
https://doi.org/10.3324/haematol.2019.231456 ©2021 Ferrata Storti Foundation
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