IBYME   02675
INSTITUTO DE BIOLOGIA Y MEDICINA EXPERIMENTAL
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Osteoclastogenesis process in bone marrow of untreated advanced breast cancer patients
Autor/es:
FERNANDEZ VALLONE, V.; CHOI, H.; MARTINEZ, L.; LAVOBSKY, V.; BATGELJ, E.; DIMASE, F.; FELDMAN, L.; BORDENAVE, R. H.; CHASSEING, N. A.
Lugar:
Philadelphia
Reunión:
Congreso; Metastasis and the tumor microenvironment; 2010
Institución organizadora:
American Association for Cancer Research
Resumen:
Osteoclastogenesis process in bone marrow of untreated
advanced breast cancer patients.
Fernández
Vallone Valeria B1, Choi Hosoon2, Martinez Leandro M1,
Labovsky Vivian1, Batgelj Emilio1, Dimase Federico1,
Feldman Leonardo1, Bordenave Raul H1, Chasseing Norma A1.
1 Institute for Experimental
Biology and Medicine, Buenos Aires, Argentina.
2
Institute
for Regenerative Medicine, Health Science Center College of Medicine, Temple,
Texas, USA.
Most of advanced breast cancer patients (BCP) develop
osteolytic bone metastasis as a result of the imbalance between osteogenesis,
osteoclastogenesis and bone resorption
processes. In previous studies we found in bone marrow (BM) aspirates of untreated BCP (infiltrative ductal breast carcinoma,
clinical stage III and IV, without bone and BM metastasis) a decrease of the
cloning efficiency of BM-mesenchymal stem cells (MSC), measured as number of
colony- forming unit-fibroblasts (CFU-F), and a decrease in its osteogenic differentiation
capacity compared to healthy volunteers (HV). It is well known that osteoblast and MSCs
regulate osteoclastogenesis, so in the present study we evaluated the
osteoclastogenic differentiation of BM-mononuclear cells (MNC) from untreated
advanced BCP without bone disease. Moreover, we studied the effect of the
conditioned medium (CM) of MNC cultures from BM-BCP over the osteoclastogenic
differentiation of HV-peripheral blood monocytes (PB-Mo). Finally, we evaluated
some stimulant (IL-6, IL-7, IL-8, IL-11, TNF-α, IL-17, PGE2, TGF-β,
soluble-RANKL, M-CSF, IL-1β, and GM-CSF) and inhibitor factors (IL-4, IL-10,
OPG and IFN-γ) of osteoclastogenesis in CM from CFU-F assays (7 and 14 days) from
BCP and HV.
Methodology:
Osteoclastic
Differentiation: a) BM-MNC
of BCP and HV: the cultures were incubated in alpha
medium suplemented with 20% horse serum with or without Vitamin D3 (VitD3)
(10-8M). After 5 days CM were harvested and cells were cultured in
the same conditions previously described. b)PB-Mo of HV: Mo were cultured in 90%
CM of BCP or HV and 10% of alpha medium supplemented with 10% FBS and M-CSF (25ng/ml).
The count of the number of osteoclast cells (OC) was made at day 16. OC= cells
with 3 or 5 nucleus, TRAP positive. Soluble factors were quantified by ELISA in
CM of CFU-F cultures (days 7 and 14) from BM of BCP and HV.
Results:
we observed spontaneous osteoclastogenesis (SpOC) in BM of BCP, (OC)%=36.9±1.9 and
non-responsiveness to VitD3, meanwhile SpOC was not observed in BM
of HV, which did respond to VitD3 (OC%=9.8±0.6). Moreover, BCP-CM treatment
induced osteoclastic differentiation of HV-PB-Mo in similar way to 25ng/ml of
sRANKL (34.6±1.9 vs 32.6±2.5, respectively). OC from patients were of a higher
size and number of nucleus/OC. In contrast, HV-CM treatment did not induced
osteoclastic differentiation of HV-PB-Mo.
SpOC in BM could be related to the major mRANKL
expression observed in MSC of BM from BCP vs HV (+++vs++). Finally, BCP
presented significantly higher levels of GM-CSF (BCP=98±4 vs. HV= <7.8 pg/ml; in 7 days CM from CFU-F) as
osteoclastogenic factor and lower levels of OPG (BCP=177±40 vs. HV=1,422± 264;
p<0.05) as
inhibitor factor. We cannot discard other soluble factors present in BM-CM from
BCP that could stimulate the differentiation of GM progenitors and PB-Mo.
Conclusion:
we consider important the evaluation of the osteoclastogenic potential of BM-
hematopoietic progenitors as well as the level of OPG and GM-CSF as possible
prognostic factors for future bone disorders that may favor the invasion of BC cells
into bone.