1.N.1 The Osteoclast Fusion Complex (OFC) Family 

Osteoclast cell fusion to form syncytia is promoted by several proteins.  Most importantly are DC-STAMP and OC-STAMP, but other proteins such as the V-type proton ATPase subunit d2, ATP6v0d2 (TC# 3.A.2.2.6), CD47, a leucocyte surface antigen of 323 aas (Q08722) and syncytin-1 (TC# 1.G.9.1.1) may play roles (Møller et al. 2016). The process is regulated by NFATc1, a master regulator of nuclear factor kappaB ligand-induced osteoclast differentiation. Kim et al. 2008 demonstrated a role for NFATc1 as a positive regulator of nuclear factor kappaB ligand-mediated osteoclast fusion as well as other fusion-inducing factors such as TNF-alpha. Exogenous overexpression of a constitutively active form of NFATc1 in bone marrow-derived monocyte/macrophage cells (BMMs) induces formation of multinucleated osteoclasts as well as the expression of fusion-mediating molecules such as the d2 isoform of vacuolar ATPase V(o) domain (Atp6v0d2) and the dendritic cell-specific transmembrane protein (DC-STAMP). Inactivation of NFATc1 by cyclosporin A treatment attenuates expression of Atp6v0d2 and DC-STAMP and the subsequent fusion of osteoclasts. Kim et al. 2008 showed that NFATc1 binds to the promoter regions of Atp6v0d2 and DC-STAMP in osteoclasts and directly induces their expression. Furthermore, overexpression of Atp6v0d2 and DC-STAMP rescues cell-cell fusion of preosteoclasts despite reduced NFATc1 activity. Thus, the NFATc1/Atp6v0d2 and DC-STAMP signaling axis plays a key role in the osteoclast multinucleation process, which is essential for efficient bone resorption.

The osteoclast is a bone-resorbing cell, which degrades bone tissues by secreting protons and lysosomal enzymes into the resorption pit. The plasma membrane facing bone tissue (ruffled border), generated partly by fusion of lysosomes, may mimic H+ flux mechanisms regulating acidic vesicles. Kuno 2018 identified three electrogenic H+-fluxes in osteoclast plasma membranes, a vacuolar H+- V-type ATPase (TC# 3.A.2), a voltage-gated proton Hv channel (TC# 1.A.51) and an acid-inducible H+-leak. The V-ATPase mediates active H+ efflux, acidifying the resorption pit, while the acid-inducible H+ leak, activated at an extracellular pH < 5.5, diminishes pit acidification, possibly to protect bone from excess degradation. The two-way H+ flux mechanisms may have advantages in fine regulation of pit pH. The Hv channel mediates passive H+ efflux. Although its working ranges are limited, the amount of H+ extrusion is 100 times larger than those of the V-ATPase and may support reactive oxygen species production during osteoclastogenesis. Extracellular Ca2+, H+ and inorganic phosphate, which accumulate in the resorption pit, either stimulate or inhibit these H+ fluxes (Kuno 2018).



Chiu, Y.H., E. Schwarz, D. Li, Y. Xu, T.R. Sheu, J. Li, K.L. de Mesy Bentley, C. Feng, B. Wang, J.C. Wang, L. Albertorio-Saez, R. Wood, M. Kim, W. Wang, and C.T. Ritchlin. (2017). Dendritic Cell-Specific Transmembrane Protein (DC-STAMP) Regulates Osteoclast Differentiation via the Ca2+ /NFATc1 Axis. J Cell Physiol 232: 2538-2549.

Kalita, M.M. and W.B. Fischer. (2017). Decoupled side chain and backbone dynamics for proton translocation - M2 of influenza A. J Mol Model 23: 212.

Kim, K., S.H. Lee, J. Ha Kim, Y. Choi, and N. Kim. (2008). NFATc1 induces osteoclast fusion via up-regulation of Atp6v0d2 and the dendritic cell-specific transmembrane protein (DC-STAMP). Mol Endocrinol 22: 176-185.

Kuno, M. (2018). Cooperative electrogenic proton transport pathways in the plasma membrane of the proton-secreting osteoclast. Pflugers Arch. [Epub: Ahead of Print]

Møller, A.M., J.M. Delaissé, and K. Søe. (2016). Osteoclast Fusion: Time-Lapse Reveals Involvement of CD47 and Syncytin-1 at Different Stages of Nuclearity. J Cell Physiol. [Epub: Ahead of Print]

Takagi, T., H. Inoue, N. Takahashi, R. Katsumata-Tsuboi, and M. Uehara. (2017). Sulforaphene attenuates multinucleation of pre-osteoclasts by suppressing expression of cell-cell fusion-associated genes DC-STAMP, OC-STAMP, and Atp6v0d2. Biosci. Biotechnol. Biochem. 81: 1220-1223.

Zhang, C., C.E. Dou, J. Xu, and S. Dong. (2014). DC-STAMP, the key fusion-mediating molecule in osteoclastogenesis. J Cell Physiol 229: 1330-1335.


TC#NameOrganismal TypeExample

The Osteoclast fusion complex including DC-STAMP (470 aas; 7 TMSs), OC-STAMP (566 aas and 7 TMSs) and Leucocyte surface antigen, CD47 of 323 aas and 5 TMSs (Chiu et al. 2016; Møller et al. 2016; Zhang et al. 2014).  CD47 is homologous to the first 5 TMSs of Presenilin-1 of Homo sapiens (1.A.54.1.1).  It is therefore a presenilin homologue (unpublished observation). Sulforaphene (SFE) downregulates mRNA expression of DC-STAMP, OC-STAMP, and Atp6v0d2, which encode cell-cell fusion molecules (Takagi et al. 2017). SFE attenuates pre-osteoclast multinucleation via suppression of cell-cell fusion. ITIM on DC-STAMP is a functional motif that regulates osteoclast differentiation through the NFATc1/Ca2+ axis (Chiu et al. 2017).

SFC of Homo sapiens