Emerging Roles of Osteoclasts in the Modulation of Bone Microenvironment and Immune Suppression in Multiple Myeloma

Mansour, Anna; Wakkach, Abdelilah; Blin-Wakkach, Claudine

doi:10.3389/fimmu.2017.00954

Cited by 34 publications

(31 citation statements)

References 121 publications

(150 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…We further determined the benefit provided by this procedure on the analysis of OCL specific properties by comparing their expression levels of well-established OCL markers. Real-time quantitative PCR analysis revealed that, compared to their bone marrow CD11b + precursor cells, sorted murine ≥3N cells expressed very high RNA levels of Tcirg1 (a3 subunit of the V-ATPAse), Calcr (calcitonin receptor), Acp5 (TRAcP), Mmp9 (matrix metalloproteinase 9), Itgb3 (Integrin-β3) and Ctsk (cathepsin K) (Figure 3 ) demonstrating that sorted multinucleated cells have the molecular signature of mature OCLs ( 3 , 20 , 21 ). Importantly, sorted 1-2N cells expressed substantially and significantly lower levels of all OCL markers than sorted mature ≥3N OCLs.…”

Section: Resultsmentioning

confidence: 99%

A Novel Reliable and Efficient Procedure for Purification of Mature Osteoclasts Allowing Functional Assays in Mouse Cells

et al. 2018

Self Cite

View full text Add to dashboard Cite

Osteoclasts (OCLs) are multinucleated phagocytes of monocytic origin responsible for physiological and pathological bone resorption including aging processes, chronic inflammation and cancer. Besides bone resorption, they are also involved in the modulation of immune responses and the regulation of hematopoietic niches. Accordingly, OCLs are the subject of an increasing number of studies. Due to their rarity and the difficulty to isolate them directly ex vivo, analyses on OCLs are usually performed on in vitro differentiated cells. In this state, however, OCLs represent a minority of differentiated cells. Since up to date a reliable purification procedure is still lacking for mature OCLs, all cells present in the culture are analyzed collectively to answer OCL-specific questions. With the development of in-depth transcriptomic and proteomic analyses, such global analyses on unsorted cells can induce severe bias effects in further results. In addition, for instance, analysis on OCL immune function requires working on purified OCLs to avoid contamination effects of monocytic precursors that may persist during the culture. This clearly highlights the need for a reliable OCL purification procedure. Here, we describe a novel and reliable method to sort OCLs based on cell multinucleation while preserving cell viability. Using this method, we successfully purified multinucleated murine cells. We showed that they expressed high levels of OCL markers and retained a high capacity of bone resorption, demonstrating that these are mature OCLs. The same approach was equally applied for the purification of human mature OCLs. Comparison of purified OCLs with mononucleated cells or unsorted cells revealed significant differences in the expression of OCL-specific markers at RNA and/or protein level. This exemplifies that substantially better outcomes for OCLs are achieved after the exclusion of mononucleated cells. Our results clearly demonstrate that the in here presented procedure for the analysis and sorting of pure OCLs represents a novel, robust and reliable method for the detailed examination of bona fide mature OCLs in a range that was previously impossible. Noteworthy, this procedure will open new perspectives into the biology of osteoclasts and osteoclast-related diseases.

show abstract

Section: Resultsmentioning

confidence: 99%

A Novel Reliable and Efficient Procedure for Purification of Mature Osteoclasts Allowing Functional Assays in Mouse Cells

et al. 2018

Self Cite

View full text Add to dashboard Cite

show abstract

“…OCs regulate the bone marrow niches for hematopoietic stem cells ( 29 ), B-cell progenitors, and the proliferation of malignant plasma cells ( 30 ). They are capable of driving immune T-cell response toward immunosuppression or inflammation according to their origin and to their environment ( 31 , 32 ), and participate in the modulation of bone microenvironment and immune suppression in multiple myeloma ( 33 , 34 ).…”

Section: Introductionmentioning

confidence: 99%

An Optimized Method to Generate Human Active Osteoclasts From Peripheral Blood Monocytes

et al. 2018

View full text Add to dashboard Cite

Osteoclasts (OCs), the bone-resorbing cells, play a key role in skeletal development and adult bone remodeling. They also participate in the pathogenesis of various bone disorders. One of the major technical difficulties in the generation of OCs, when working on human material, is the ability to achieve large differentiation of mature OCs from human peripheral blood mononuclear cells (PBMCs). Access to a standardized source of active OCs is needed to better analyze the roles of human OCs. The aim of this study was to develop a procedure yielding active and mature OCs from fresh human PBMCs. We therefore examined the differentiation of PBMCs to OCs in different cell culture media, using non-stripped and charcoal-stripped sera in the presence of macrophage colony-stimulating factor (M-CSF) and receptor activator of nuclear factor kappa-B ligand (RANKL). We also studied the effects of vitamin D3 in the differentiation level of PBMCs to OCs. Phalloidin-AlexaFluor®488/DAPI fluorescent stainings and dentin resorption analyses by scanning electron microscopy were used to identify the number and size of differentiated OCs, number of nuclei per cell and resorption activities of OCs for a 7–14–21-day culture period. This study reports an optimized method for an efficient production of human active OCs from a low seeding density of PBMCs, after a 14-day culture period by using a medium containing fetal bovine charcoal-stripped serum in the presence of M-CSF and RANKL, and in the absence of vitamin D3.

show abstract

“…MM is characterized by the production of monoclonal intact immunoglobulins or immunoglobulin free light chains, leading on to renal failure, anemia, immunosuppression, and osteolytic bone disease, that occurs in most of these patients ( 3 ). The latter is, indeed, detected in ~70% of cases at the initial diagnosis of MM; it persists even in the absence of active disease, being a major cause of morbidity and mortality in MM patients ( 4 ). Despite the improvement in treatment options, during the clinical course of the disease 80–90% of patients experience skeletal-related events (SRE) such as bone pain (70–80% of the patients), spontaneous fractures (50–60%), hypercalcemia (15%), and spinal cord compression (2–3%) ( 4 ).…”

Section: Introductionmentioning

confidence: 99%

“…The latter is, indeed, detected in ~70% of cases at the initial diagnosis of MM; it persists even in the absence of active disease, being a major cause of morbidity and mortality in MM patients ( 4 ). Despite the improvement in treatment options, during the clinical course of the disease 80–90% of patients experience skeletal-related events (SRE) such as bone pain (70–80% of the patients), spontaneous fractures (50–60%), hypercalcemia (15%), and spinal cord compression (2–3%) ( 4 ). The development of bone lytic lesions is due to an alteration of the dynamic balance between bone-resorption and bone-formation, caused by an enhanced OC formation and activity, and an impaired osteoblast function ( 5 , 6 ).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

LIGHT/TNFSF14 as a New Biomarker of Bone Disease in Multiple Myeloma Patients Experiencing Therapeutic Regimens

et al. 2018

View full text Add to dashboard Cite

We have previously shown that through the production of high LIGHT levels, immune cells contribute to both osteoclastogenesis and bone destruction in Multiple Myeloma (MM)-related bone disease. With the aim of further exploring the mechanisms underlying the development of MM-related bone disease, here we focused on a possible role of LIGHT in MM patients with active bone disease despite the treatment received. We detected LIGHT over-expression by circulating CD14+ monocytes from MM patients still showing active bone disease, despite the treatment. In addition, we found over-expression of receptor activator of nuclear factor kappa-B ligand (RANKL), whose pro-osteoclastogenic role is well-known, in T-lymphocytes isolated from the same patients. Although the percentages of circulating osteoclast progenitors, CD14+CD16+ monocytes, were higher in all the MM patients than in the controls spontaneous osteoclastogenesis occurred only in the cultures derived from PBMCs of MM patients with unresponsive bone disease. Of note, in the same cultures osteoclastogenesis was partially or completely inhibited, in a dose-dependent manner, by the addition of RANK-Fc or anti-LIGHT neutralizing antibody, demonstrating the contribution of both LIGHT and RANKL to the enhanced osteoclast formation observed. In addition, high serum levels of TRAP5b and CTX, the two markers of osteoclast activity, were detected in MM patients with bone disease not responsive to treatment. In conclusion, our study indicates a prominent role of LIGHT in the crosstalk among osteoclasts and immune cells, co-involved together with RANKL in the pathophysiological mechanisms leading to MM-related bone disease. This TNF superfamily member may thus be a possible new therapeutic target in MM-related bone disease.

show abstract

Emerging Roles of Osteoclasts in the Modulation of Bone Microenvironment and Immune Suppression in Multiple Myeloma

Cited by 34 publications

References 121 publications

A Novel Reliable and Efficient Procedure for Purification of Mature Osteoclasts Allowing Functional Assays in Mouse Cells

A Novel Reliable and Efficient Procedure for Purification of Mature Osteoclasts Allowing Functional Assays in Mouse Cells

An Optimized Method to Generate Human Active Osteoclasts From Peripheral Blood Monocytes

LIGHT/TNFSF14 as a New Biomarker of Bone Disease in Multiple Myeloma Patients Experiencing Therapeutic Regimens

Contact Info

Product

Resources

About