Induction of osteoclastogenesis in an in vitro model of Gaucher disease is mediated by T cells via TNF-α

LISIN, Departamento de Ciencias Biológicas, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata (1900) Argentina.
Gene (Impact Factor: 2.14). 11/2012; 509(1):51-9. DOI: 10.1016/j.gene.2012.07.071
Source: PubMed


Gaucher disease is a lysosomal storage disorder caused by deficiency of glucocerebrosidase enzymatic activity leading to accumulation of its substrate glucocerebrosidase mainly in macrophages. Skeletal disorder of Gaucher disease is the major cause of morbidity and is highly refractory to enzyme replacement therapy. However, pathological mechanisms of bone alterations in Gaucher disease are still poorly understood. We hypothesized that cellular alteration in Gaucher disease produces a proinflammatory milieu leading to bone destruction through enhancement of monocyte differentiation to osteoclasts and osteoclasts resorption activity. Against this background we decided to investigate in an in vitro chemical model of Gaucher disease, the capacity of secreted soluble mediators to induce osteoclastogenesis, and the mechanism responsible for this phenomena. We demonstrated that soluble factors produced by CBE-treated PBMC induced differentiation of osteoclasts precursors into mature and active osteoclasts that express chitotriosidase and secrete proinflammatory cytokines. We also showed a role of TNF-α in promoting osteoclastogenesis in Gaucher disease chemical model. To analyze the biological relevance of T cells in osteoclastogenesis of Gaucher disease, we investigated this process in T cell-depleted PBMC cultures. The findings suggest that T cells play a role in osteoclast formation in Gaucher disease. In conclusion, our data suggests that in vitro GCASE deficiency, along with concomitant glucosylceramide accumulation, generates a state of osteoclastogenesis mediated in part by pro-resorptive cytokines, especially TNF-α. Moreover, T cells are involved in osteoclastogenesis in Gaucher disease chemical model.

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Available from: Victoria Delpino, Sep 24, 2014
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    • "Interestingly, although CHIA also possesses chitinolytic activity and plays a critical role in tissue remodeling [36], its expression in osteoclast differentiation resulted undetectable. CHI3L1 had been linked to tissue remodeling [17] [37], joint injury [38], and significantly elevated levels of CHI3L1 protein have been detected in serum and synovial fluid from OA patients [39] [40]. The plethora of evidences showing that CHI3L1 stimulates proliferation of connective tissue cells and modulates expression levels of chemokines and metalloproteases in inflammatory fibroblasts, and that enhances chemotaxis of endothelial cells [41] [42] strongly indicate that CHI3L1 plays crucial role in stromal cells not only in inflammatory conditions. "
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    ABSTRACT: Mammalian Chitinases consisting of CHIA, CHIT1, CHI3L1, CHI3L2 and CHID1 exerts important biological roles in the monocyte lineage and chronic inflammatory diseases. Pathological bone resorption is a cause of significant morbidity in diseases affecting the skeleton such as rheumatoid arthritis, osteoporosis, periodontitis and cancer metastasis. The biologic role of Chitinases in bone resorption is poorly understood. In this study, we evaluated the expression of the Chitinases family during osteoclast differentiation. The expression of CHIA, CHI3L2 and CHID1 resulted unchanged during osteoclast differentiation, whereas CHIT1 and CHI3L1 increased significantly. We also observed thatCHIT1 and CHI3L1 are involved in osteoclast function. Indeed, silencing CHIT1 and CHI3L1 with siRNA resulted in a significant decrease in bone resorption activity. In addition, transfection with CHIT1 or CHI3L1 siRNA and co-transfection with both, decreased the levels of the pro-differentiative marker MMP9. Overall, these discoveries reveal a novel and crucial role for both CHIT1 and CHI3L1 in promoting bone resorption and identify new potential candidate markers for therapeutic targeting.
    Full-text · Article · Jan 2014 · Bone
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    • "These cells were cultured in AIM-V media (Invitrogen, Carlsbad, CA, USA) at 37 °C in a 5% CO 2 atm to a number of 10 6 cells/ml for 72 h in the presence or absence of conduritolbeta-epoxide (CBE) 500 mM (Sigma, St Louis, MO, USA). The effect of CBE was tested, demonstrating the inhibition of glucocerebrosidase activity and glucocerebroside accumulation, as previously described (Mucci et al., 2012). Conditioned media (CM) was obtained by centrifugation of the cultures. "
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    ABSTRACT: Gaucher disease (GD) is caused by mutations in GBA gene that confer a deficient level of activity of glucocerebrosidase (GCase). This deficiency leads to accumulation of the glycolipid glucocerebroside in the lysosomes of cells of monocyte/macrophage system. Type I GD is the mildest form and is characterized by the absence of neuronopathic affection. Bone compromise in Gaucher disease patients is the most disabling aspect of the disease. However, pathophysiological aspects of skeletal alterations are still poorly understood. The homeostasis of bone tissue is maintained by the balanced processes of bone resorption by osteoclasts and formation by osteoblasts. We decided to test weather bone resorption and/or bone formation could be altered by the use of a chemical in vitro murine model of Gaucher disease. We used two sources of cells from monocyte/macrophages lineage isolated from normal mice, splenocytes (S) and peritoneal macrophages (PM), and were exposed to CBE, the inhibitor of GCase (S-CBE and PM-CBE, respectively). Addition of both conditioned media (CM) from S-CBE and PM-CBE induced the differentiation of osteoclasts precursors from bone marrow to mature and functional osteoclasts. TNF-α could be one of the factors responsible for this effect. On the other side, addition of CM to an osteoblast cell culture resulted in a reduction in expression of alkaline phosphatase and mineralization process. In conclusion, these results suggest implication of changes in both bone formation and bone resorption and are consistent with the idea that both sides of the homeostatic balance are affected in GD.
    Full-text · Article · Sep 2013 · Gene
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    • "Bone mass homeostasis is also dependant on osteoclasts activity. It has been shown, in an vitro CBE chemical model, that conditioned media from CBE-treated PBMC induced ex vivo osteoclastogenesis [28]. We quantified the resorbing potential of CD14+-derived osteoclasts cultured in osteoclastic media on calcium phosphate matrix. "
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    ABSTRACT: Gaucher disease (GD) is an autosomal recessive disorder characterized by lysosomal glucocerebrosidase (GBA) deficiency leading to hematological and skeletal manifestations. Mechanisms underlying these symptoms have not yet been elucidated. In vivo, bone marrow (BM) mesenchymal stem cells (MSCs) have important role in the regulation of bone mass and in the support of hematopoiesis, thus representing potential candidate that could contribute to the disease. GBA deficiency may also directly impair hematopoietic stem/progenitors cells (HSPCs) intrinsic function and induce hematological defect. In order to evaluate the role of BM stem cells in GD pathophysiology, we prospectively analyzed BM-MSCs and HSPCs properties in a series of 10 patients with type 1 GD. GBA activity was decreased in all tested cell subtypes. GD-MSCs had an impaired growth potential, morphological and cell cycle abnormalities, decreased capacities to differentiate into osteoblasts. Moreover, GD-MSCs secreted soluble factors that stimulated osteoclasts resorbing activities. In vitro and in vivo primitive and mature hematopoiesis were similar between patients and controls. However, GD-MSCs had a lower hematopoietic supportive capacity than those from healthy donors. These data suggest that BM microenvironment is altered in GD and that MSCs are key components of the manifestations observed in GD.
    Full-text · Article · Jul 2013 · PLoS ONE
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