Osteoclast-derived apoptotic bodies couple bone resorption and formation in bone remodeling

Abstract

Bone remodeling is precisely coordinated by bone resorption and formation. Apoptotic osteoclasts generate large amounts of apoptotic bodies (ABs) marking the end of the bone resorption phase, whereas the functions of osteoclast-derived ABs remain largely unknown. Here, we identified the molecular profile of ABs derived from osteoclasts at distinct differentiation stages and investigated their corresponding functions. ABs were isolated from apoptotic bone marrow macrophages, preosteoclasts, and mature osteoclasts induced by staurosporine. Proteomic signature analysis with liquid chromatography-tandem mass spectrometry suggested marked protein cargo differences among the different ABs. Further bioinformatic analysis showed that the proteomic signatures of the ABs were highly similar to those of their parental cells. Functionally, pOC-ABs induced endothelial progenitor cell differentiation and increased CD31^hiEmcn^hi endothelial cell formation in a murine bone defect model via their PDGF-BB cargo. mOC-ABs induced osteogenic differentiation of mesenchymal stem cells and facilitated osteogenesis via RANKL reverse signaling. In summary, we mapped the detailed proteomic landscapes of ABs derived from osteoclasts and showed that their potential biological roles are important in coupling bone formation with resorption during bone remodeling.

References

[1] Steller H. Mechanisms and genes of cellular suicide. Science, 1995, 267:1445-1449

[2] Croft DR et al. Actin-myosin-based contraction is responsible for apoptotic nuclear disintegration. J. Cell Biol., 2005, 168:245-255

[3] Atkin-Smith GK et al. A novel mechanism of generating extracellular vesicles during apoptosis via a beads-on-a-string membrane structure. Nat. Commun., 2015, 6

[4] Zaborowski MP, Balaj L, Breakefield XO, Lai CP. Extracellular vesicles: composition, biological relevance, and methods of study. Bioscience, 2015, 65:783-797

[5] Poon IK et al. Unexpected link between an antibiotic, pannexin channels and apoptosis. Nature, 2014, 507:329-334

[6] Wickman G, Julian L, Olson MF. How apoptotic cells aid in the removal of their own cold dead bodies. Cell Death Differ., 2012, 19:735-742

[7] Zernecke A et al. Delivery of microRNA-126 by apoptotic bodies induces CXCL12-dependent vascular protection. Sci. Signal., 2009, 2:ra81

[8] Bergsmedh A et al. Horizontal transfer of oncogenes by uptake of apoptotic bodies. Proc. Natl Acad. Sci. USA, 2001, 98:6407-6411

[9] Schiller M et al. Autoantigens are translocated into small apoptotic bodies during early stages of apoptosis. Cell Death Differ., 2008, 15:183-191

[10] Akers JC et al. Biogenesis of extracellular vesicles (EV): exosomes, microvesicles, retrovirus-like vesicles, and apoptotic bodies. J. Neurooncol., 2013, 113:1-11

[11] Berda-Haddad Y et al. Sterile inflammation of endothelial cell-derived apoptotic bodies is mediated by interleukin-1alpha. Proc. Natl Acad. Sci. USA, 2011, 108:20684-20689

[12] Caruso S, Poon IKH. Apoptotic cell-derived extracellular vesicles: more than just debris. Front Immunol., 2018, 9:1486

[13] Boyle WJ, Simonet WS, Lacey DL. Osteoclast differentiation and activation. Nature, 2003, 423:337-342

[14] Yasuda H et al. Osteoclast differentiation factor is a ligand for osteoprotegerin/osteoclastogenesis-inhibitory factor and is identical to TRANCE/RANKL. Proc. Natl Acad. Sci. USA, 1998, 95:3597-3602

[15] Xie H et al. PDGF-BB secreted by preosteoclasts induces angiogenesis during coupling with osteogenesis. Nat. Med., 2014, 20:1270-1278

[16] Ikebuchi Y et al. Coupling of bone resorption and formation by RANKL reverse signalling. Nature, 2018, 561:195-200

[17] Ma Q et al. Mature osteoclast-derived apoptotic bodies promote osteogenic differentiation via RANKL-mediated reverse signaling. J. Biol. Chem., 2019, 294:11240-11247

[18] Dou C et al. Changing expression profiles of lncRNAs, mRNAs, circRNAs and miRNAs during osteoclastogenesis. Sci. Rep., 2016, 6

[19] Marks SC Jr., Seifert MF. The lifespan of osteoclasts: experimental studies using the giant granule cytoplasmic marker characteristic of beige mice. Bone, 1985, 6:451-455

[20] Manolagas SC. Birth and death of bone cells: basic regulatory mechanisms and implications for the pathogenesis and treatment of osteoporosis. Endocr. Rev., 2000, 21:115-137

[21] Brandi ML, Collin-Osdoby P. Vascular biology and the skeleton. J. Bone Miner. Res., 2006, 21:183-192

[22] Chim SM et al. Angiogenic factors in bone local environment. Cytokine Growth Factor Rev., 2013, 24:297-310

[23] Zhu S et al. Coupling factors and exosomal packaging microRNAs involved in the regulation of bone remodelling. Biol. Rev. Camb. Philos. Soc., 2018, 93:469-480

[24] Li D et al. Osteoclast-derived exosomal miR-214-3p inhibits osteoblastic bone formation. Nat. Commun., 2016, 7

[25] Huynh N et al. Characterization of regulatory extracellular vesicles from osteoclasts. J. Dent. Res., 2016, 95:673-679

[26] Belmokhtar CA, Hillion J, Segal-Bendirdjian E. Staurosporine induces apoptosis through both caspase-dependent and caspase-independent mechanisms. Oncogene, 2001, 20:3354-3362

[27] Atkin-Smith GK et al. Isolation of cell type-specific apoptotic bodies by fluorescence-activated cell sorting. Sci. Rep., 2017, 7

[28] Tyanova S, Temu T, Cox J. The MaxQuant computational platform for mass spectrometry-based shotgun proteomics. Nat. Protoc., 2016, 11:2301-2319

[29] Bar-Shavit Z. The osteoclast: a multinucleated, hematopoietic-origin, bone-resorbing osteoimmune cell. J. Cell. Biochem., 2007, 102:1130-1139

[30] Kusumbe AP, Ramasamy SK, Adams RH. Coupling of angiogenesis and osteogenesis by a specific vessel subtype in bone. Nature, 2014, 507:323-328

[31] Ding N et al. Alendronate induces osteoclast precursor apoptosis via peroxisomal dysfunction mediated ER stress. J. Cell. Physiol., 2018, 233:7415-7423

[32] Robbins PD, Morelli AE. Regulation of immune responses by extracellular vesicles. Nat. Rev. Immunol., 2014, 14:195-208

[33] Buzas EI et al. Emerging role of extracellular vesicles in inflammatory diseases. Nat. Rev. Rheumatol., 2014, 10:356-364

[34] Schorey JS, Cheng Y, Singh PP, Smith VL. Exosomes and other extracellular vesicles in host-pathogen interactions. EMBO Rep., 2015, 16:24-43

[35] Zhu Z et al. Macrophage-derived apoptotic bodies promote the proliferation of the recipient cells via shuttling microRNA-221/222. J. Leukoc. Biol., 2017, 101:1349-1359

[36] Becker A et al. Extracellular vesicles in cancer: cell-to-cell mediators of metastasis. Cancer Cell, 2016, 30:836-848

[37] Furlan A et al. Ets-1 drives breast cancer cell angiogenic potential and interactions between breast cancer and endothelial cells. Int. J. Oncol., 2019, 54:29-40

[38] Viallard C, Larrivee B. Tumor angiogenesis and vascular normalization: alternative therapeutic targets. Angiogenesis, 2017, 20:409-426

[39] Zhou W et al. Cancer-secreted miR-105 destroys vascular endothelial barriers to promote metastasis. Cancer Cell, 2014, 25:501-515

[40] Hoshino A et al. Tumour exosome integrins determine organotropic metastasis. Nature, 2015, 527:329-335

[41] Liu M, Sun Y, Zhang Q. Emerging role of extracellular vesicles in bone remodeling. J. Dent. Res., 2018, 97:859-868

[42] Xiao Z et al. Analysis of the extracellular matrix vesicle proteome in mineralizing osteoblasts. J. Cell. Physiol., 2007, 210:325-335

[43] Deng L et al. Osteoblast-derived microvesicles: a novel mechanism for communication between osteoblasts and osteoclasts. Bone, 2015, 79:37-42

[44] Qin Y et al. Myostatin inhibits osteoblastic differentiation by suppressing osteocyte-derived exosomal microRNA-218: a novel mechanism in muscle-bone communication. J. Biol. Chem., 2017, 292:11021-11033

[45] Chen C et al. Mesenchymal stem cell transplantation in tight-skin mice identifies miR-151-5p as a therapeutic target for systemic sclerosis. Cell Res., 2017, 27:559-577

[46] Tang Y et al. TGF-beta1-induced migration of bone mesenchymal stem cells couples bone resorption with formation. Nat. Med., 2009, 15:757-765

[47] Su J et al. Phylogenetic distinction of iNOS and IDO function in mesenchymal stem cell-mediated immunosuppression in mammalian species. Cell Death Differ., 2014, 21:388-396

[48] Ma Q et al. Osteoclast-derived apoptotic bodies show extended biological effects of parental cell in promoting bone defect healing. Theranostics, 2020, 10:6825-6838

[49] Yu G, Wang LG, Han Y, He QY. clusterProfiler: an R package for comparing biological themes among gene clusters. OMICS, 2012, 16:284-287

[50] Dossa T et al. Osteoclast-specific inactivation of the integrin-linked kinase (ILK) inhibits bone resorption. J. Cell. Biochem., 2010, 110:960-967

Funding

National Natural Science Foundation of China (National Science Foundation of China)(81802166)

Nature Science Foundation of China (81802166) first class General Financial Grant from the China Postdoctoral Science Foundation (2017M613315) TMMU funding for young investigators (2017MPRC-04)