miR-29b negatively regulates human osteoclastic cell differentiation and function: Implications for the treatment of multiple myeloma-related bone disease

2013 ◽  
Vol 228 (7) ◽  
pp. 1506-1515 ◽  
Author(s):  
Marco Rossi ◽  
Maria Rita Pitari ◽  
Nicola Amodio ◽  
Maria Teresa Di Martino ◽  
Francesco Conforti ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Cell Differentiation ◽  
Bone Disease ◽  
And Function ◽  
Osteoclastic Cell

Dasatinib Promotes Osteoprogenitor Differentiation and Inhibition of Osteoclastogenesis: Rationale for Treatment of Myeloma Bone Disease.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 3834-3834
Author(s):  
Antonio Garcia-Gomez ◽  
Mercedes Garayoa ◽  
Enrique M. Ocio ◽  
Edvan Crusoe ◽  
Diego Fernández ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Cell Lines ◽  
Bone Disease ◽  
Plasma Cells ◽  
Annexin V ◽  
Conditioned Media ◽  
Tartrate Resistant Acid Phosphatase ◽  
Alizarin Red ◽  
Alp Activity ◽  
And Function

Abstract Abstract 3834 Poster Board III-770 Introduction Multiple myeloma (MM), an hematological malignancy of terminally differentiated plasma cells, is characterized by the presence of bone disease, caused by increased osteoclast (OC) activity and differentiation as well as a reduction in osteoblast (OB) number and function. Dasatinib (BMS-354825) is an oral multitargeted tyrosin-kinase inhibitor approved for chronic myeloid leukemia which is also under clinical investigation in several other types of tumors. Preclinical data suggests that dasatinib can also be of value in MM based on its effects on myelomatous plasma cells and angiogenesis. In this study, we have further investigated the effects of dasatinib on in vitro OB differentiation and function, as well as on OC formation and resorption activity. Materials and methods For studies on OB differentiation and function, human mensenchymal stem cells (hMSC) from bone marrow samples of healthy donors and MM patients were used. Alternatively, the mesenchymal hMSC-TERT, the osteoblast-like (MG-63) and multiple myeloma (MM.1S) cell lines were employed. Dasatinib mechanism of action was investigated by Western blotting, PKH67/Annexin V/7 aminoactinomycin D staining, real time RT-PCR, alkaline phosphatase (ALP) activity and quantitative mineralization assays. Receptor activator of nuclear factor κ B ligand (RANKL) and osteoprotegerin (OPG) levels in conditioned media were measured by ELISA. OCs were generated by culture of peripheral blood mononuclear cells from healthy volunteers in medium containing macrophage colony stimulating factor and RANKL. The effect of dasatinib on osteoclastogenesis was assessed by enumeration of multinucleated (≥3) tartrate resistant acid phosphatase-positive cells, whereas bone resorption was calculated by the resorbed area on calcium-coated slides. Results We found that dasatinib inhibited platelet derived growth factor (PDGF)-stimulated activation of PDGF receptor β (PDGFRβ) and c-Src in hMSC-TERT and MG-63 cell lines, both tyrosin kinases implicated in OB remodelation processes. Inhibition of PDGFRβ and c-Src signalling correlated with diminished proliferation of the same cell lines without affecting cell viavility as assessed by PKH67/Annexin V/7 aminoactinomycin D staining. Also, treatment of human osteoprogenitor cells with low dasatinib concentrations (2 - 5 nM) promoted OB differentiation since ALP activity at day 7 and gene expression levels of bone formation markers (Runx2/Cbfa1, ALP, and COLIA1) at day 7 and 14 in the osteoblastic differentiation process, were elevated; besides, dasatinib also increased mineral nodular formation as per quantification of alizarin red staining. Finally, treatment with dasatinib decreased the RANKL/OPG ratio in conditioned media from co-cultures of MG-63 and MM.1S cell lines. Similar range of dasatinib concentrations markedly inhibited OC formation, both at the initial and late stages of differentiation from hemopoietic progenitor cells. Finally, a significant reduction of OC resorptive activity of a calcium-coated substrate was observed. Conclusion Our results indicate that dasatinib favours both OB differentiation and activity and markedly impairs osteoclastogenesis and OC resorption, thus providing rationale for its use to improve bone lesions in MM patients and other bone pathologies. This work was supported by grants from Ministerio de Ciencia e Innovación - ISCIII (PI081825); Fundación de Investigación Médica Mutua Madrileña AP27262008, and Centro en Red de Medicina Regenerativa y Terapia Celular de Castilla y León 07-09, Consejería Sanidad JCyL-ISCIII. Disclosures: Garzon: Bristol-Myers Squibb Company: Employment.

Mir-29b Negatively Regulates Osteoclastic Cell Differentiation and Function: Implications for the Treatment of Multiple Myeloma-Related Bone Disease

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 3960-3960 ◽  
Author(s):  
Marco Rossi ◽  
Maria Rita Pitari ◽  
Nicola Amodio ◽  
Maria Teresa Di Martino ◽  
Emanuela Leone ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Bone Disease ◽  
Functional Data ◽  
Conflicts Of Interest ◽  
Lentiviral Vector ◽  
Statistical Significance ◽  
Constitutive Expression ◽  
Type I ◽  
Final Maturation ◽  
And Function

Abstract Abstract 3960 Overwhelming osteoclast (OCL) activation plays a central role in multiple myeloma (MM)-related bone disease. It is well known that in MM, OCL differentiation and final maturation rely upon RANKL stimulation by bone marrow mesenchymal cells and osteoblasts (OBLs). The clinical relevance of this pathway has been recently underlined by the anti-resorptive activity mediated by the mAb Denosumab. Since the discovery of microRNAs (miRNAs), findings on their role on intracellular pathway control have undergone a tremendous progress suggesting their potential use as a therapeutic tool against specific targets. Based on these premises, we aimed to identify miRNAs that can be relevant for the management of MM-related bone disease. Among miRNAs deregulated in MM, miR-29 family has been implicated in bone pathophysiology. Indeed, miR-29 family promotes OBL generation, while miR-29b might interfere with OCL differentiation and function by targeting RANKL axis and Metalloproteinase II (MMP2), that confers the property to degradate type I collagens. Therefore, we studied whether miR-29b can have a detrimental effect for terminally differentiated OCLs. We generated OCLs in vitro upon RANKL/MCSF stimulation starting from CD14+ hematopoietic precursors and found that miR-29b levels decline along OCL differentiation, reaching statistical significance as compared with its precursors (p=0,013). These data suggested that terminally differentiated OCLs do not require miR-29b. In order to assess whether miR-29b reconstitution could affect OCL activity, we transduced OCLs with a miR-29b coding sequence- carrying lentiviral vector (29b OCLs) or an empty lentiviral vector (eOCLs) and evaluated them in morphological and functional assays. As we expected, 29b OCLs showed a faint and irregular expression of tartrate acid phosphatase (TRAcP), which is highly and uniformously distributed within WT/eOCLs. Furthermore, when 29b OCLs were seeded on dentin, generation of lacunae on dentin surface, which recapitulates bone resorption, was significantly reduced as compared with WT/eOCLs (see Figure). According to these results, we measured the release of type I collagen fragments by OCLs and found that degradation of type I collagens was significantly impaired in 29b OCLs, suggesting that constitutive expression of miR-29b strongly antagonizes OCL differentiation and bone lytic functions. In order to reproduce the features of MM related bone disease, we co-cultured OCLs with RPMI-8226 MM cells, which are able to stimulate WT/eOCLs to generate lacunae on dentin slices in the absence of exogenous RANKL/MCSF. We observed that 29b OCLs failed to generate comparable numbers and areas of the pits in presence of MM cells (p=0,035/p=0,04). To support these functional data and ascertain the effects of miR-29b on specific pathways, we evaluated the expression of SP1 and NFATc-1, which are relevant transcription factors for OCL differentiation and work through RANK-L axis. We found that both factors are down-modulated in 29b OCLs as compared to WT/eOCLs. Down modulation was observed also for MMP2, thus mirroring the reduced capability to lyse type I collagens. Overall, our data indicate that miR-29b impairs OCL differentiation and function even in presence of robust stimuli such as RANKL and MCSF. We provided molecular support to these functional data, showing that SP1 and NFATc are down modulated in presence of miR-29b as well as MMP2, which is involved in collagen degradation. Intriguingly, MM cells, which represent a strong pro-osteoclastic factor, were not able to revert OCL functional impairment. We believe that these relevant preclinical findings allow to propose miR-29b mimics as a suitable and attractive candidate to be developed as a novel and innovative treatment of MM-related bone disease. Disclosures: No relevant conflicts of interest to declare.

Involvement of LIGHT in multiple myeloma bone disease

2014 ◽  
Author(s):  
Angela Oranger ◽  
Giacomina Brunetti ◽  
Giorgio Mori ◽  
Claudia Carbone ◽  
Isabella Gigante ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Bone Disease ◽  
Myeloma Bone Disease

Anti-sclerostin treatment prevents multiple myeloma bone disease and reduces tumour burden

2016 ◽  
Author(s):  
Michelle McDonald ◽  
Michaela Reagan ◽  
Rachael Terry ◽  
Jessica Pettitt ◽  
Lawrence Le ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Bone Disease ◽  
Tumour Burden ◽  
Myeloma Bone Disease

Contribution of multiple myeloma-derived exosomes to bone disease

2016 ◽  
Author(s):  
Lavinia Raimondi ◽  
Luca Angela De ◽  
Valeria Carina ◽  
Valentina Agnese ◽  
Simona Fontana ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Bone Disease

Improved Beta-Cell Differentiation and Function in Isolated Islets Established in Dynamic Culture to Reduce Hypoxia

Diabetes ◽  
2018 ◽  
Vol 67 (Supplement 1) ◽  
pp. 303-LB
Author(s):  
NAJWA A. AL-JAHDHAMI ◽  
SCOTT J. ANDERSON ◽  
ALI ALDIBBIAT ◽  
JAMES A. SHAW
Keyword(s):  
Cell Differentiation ◽  
Beta Cell ◽  
Isolated Islets ◽  
Dynamic Culture ◽  
And Function

Bone disease as the first manifestation of systemic AL-amyloidosis

2020 ◽  
Vol 92 (7) ◽  
pp. 85-89
Author(s):  
L. P. Mendeleeva ◽  
I. G. Rekhtina ◽  
A. M. Kovrigina ◽  
I. E. Kostina ◽  
V. A. Khyshova ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Bone Marrow ◽  
Bone Disease ◽  
Plasma Cells ◽  
Interventricular Septum ◽  
Bone Destruction ◽  
Amyloid Deposition ◽  
Bone Lesions ◽  
Al Amyloidosis ◽  
Linear Type

Our case demonstrates severe bone disease in primary AL-amyloidosis without concomitant multiple myeloma. A 30-year-old man had spontaneous vertebral fracture Th8. A computed tomography scan suggested multiple foci of lesions in all the bones. In bone marrow and resected rib werent detected any tumor cells. After 15 years from the beginning of the disease, nephrotic syndrome developed. Based on the kidney biopsy, AL-amyloidosis was confirmed. Amyloid was also detected in the bowel and bone marrow. On the indirect signs (thickening of the interventricular septum 16 mm and increased NT-proBNP 2200 pg/ml), a cardial involvement was confirmed. In the bone marrow (from three sites) was found 2.85% clonal plasma cells with immunophenotype СD138+, СD38dim, СD19-, СD117+, СD81-, СD27-, СD56-. FISH method revealed polysomy 5,9,15 in 3% of the nuclei. Serum free light chain Kappa 575 mg/l (/44.9) was detected. Multiple foci of destruction with increased metabolic activity (SUVmax 3.6) were visualized on PET-CT, and an surgical intervention biopsy was performed from two foci. The number of plasma cells from the destruction foci was 2.5%, and massive amyloid deposition was detected. On CT scan foci of lesions differed from bone lesions at multiple myeloma. Bone fragments of point and linear type (button sequestration) were visualized in most of the destruction foci. The content of the lesion was low density. There was no extraossal spread from large zones of destruction. There was also spontaneous scarring of the some lesions (without therapy). Thus, the diagnosis of multiple myeloma was excluded on the basis based on x-ray signs, of the duration of osteodestructive syndrome (15 years), the absence of plasma infiltration in the bone marrow, including from foci of bone destruction by open biopsy. This observation proves the possibility of damage to the skeleton due to amyloid deposition and justifies the need to include AL-amyloidosis in the spectrum of differential diagnosis of diseases that occur with osteodestructive syndrome.

Sign in / Sign up

Export Citation Format

Share Document