Annexin A5 derived from matrix vesicles protects against osteoporotic bone loss via mineralization

Abstract

Matrix vesicles (MVs) have shown strong effects in diseases such as vascular ectopic calcification and pathological calcified osteoarthritis and in wound repair of the skeletal system due to their membranous vesicle characteristics and abundant calcium and phosphorus content. However, the role of MVs in the progression of osteoporosis is poorly understood. Here, we report that annexin A5, an important component of the matrix vesicle membrane, plays a vital role in bone matrix homeostasis in the deterioration of osteoporosis. We first identified annexin A5 from adherent MVs but not dissociative MVs of osteoblasts and found that it could be sharply decreased in the bone matrix during the occurrence of osteoporosis based on ovariectomized mice. We then confirmed its potential in mediating the mineralization of the precursor osteoblast lineage via its initial binding with collagen type I to achieve MV adhesion and the subsequent activation of cellular autophagy. Finally, we proved its protective role in resisting bone loss by applying it to osteoporotic mice. Taken together, these data revealed the importance of annexin A5, originating from adherent MVs of osteoblasts, in bone matrix remodeling of osteoporosis and provided a new strategy for the treatment and intervention of bone loss.

References

[1] Compston JE, McClung MR, Leslie WD. Osteoporosis. Lancet, 2019, 393: 364-376

[2] Estell EG, Rosen CJ. Emerging insights into the comparative effectiveness of anabolic therapies for osteoporosis. Nat. Rev. Endocrinol., 2021, 17: 31-46

[3] Chotiyarnwong P, McCloskey EV. Pathogenesis of glucocorticoid-induced osteoporosis and options for treatment. Nat. Rev. Endocrinol., 2020, 16: 437-447

[4] Bolean M et al. Biophysical aspects of biomineralization. Biophys. Rev., 2017, 9: 747-760

[5] Proudfoot D. Calcium signaling and tissue calcification. Cold Spring Harb. Perspect. Biol., 2019, 11: a035303

[6] New SE et al. Macrophage-derived matrix vesicles: an alternative novel mechanism for microcalcification in atherosclerotic plaques. Circ. Res., 2013, 113: 72-77

[7] Chen NX, O’Neill KD, Moe SM. Matrix vesicles induce calcification of recipient vascular smooth muscle cells through multiple signaling pathways. Kidney Int., 2018, 93: 343-354

[8] Bobryshev YV et al. Are calcifying matrix vesicles in atherosclerotic lesions of cellular origin? Basic Res. Cardiol., 2007, 102: 133-143

[9] Iwayama T et al. Matrix vesicle-mediated mineralization and potential applications. J. Dent. Res., 2022, 101: 1554-1562

[10] Anderson HC. Electron microscopic studies of induced cartilage development and calcification. J. Cell Biol., 1967, 35: 81-101

[11] Bonucci E. Fine structure of early cartilage calcification. J. Ultrastruct. Res., 1967, 20: 33-50

[12] Anderson HC. Vesicles associated with calcification in the matrix of epiphyseal cartilage. J. Cell Biol., 1969, 41: 59-72

[13] Boyan BD et al. The role of matrix-bound extracellular vesicles in the regulation of endochondral bone formation. Cells, 2022, 11: 1619

[14] Garces-Ortiz M, Ledesma-Montes C, Reyes-Gasga J. Presence of matrix vesicles in the body of odontoblasts and in the inner third of dentinal tissue: a scanning electron microscopic study. Med. Oral. Patol. Oral. Cir. Bucal, 2013, 18: e537-e541

[15] Shapiro IM, Landis WJ, Risbud MV. Matrix vesicles: are they anchored exosomes? Bone, 2015, 79: 29-36

[16] Bottini M et al. Matrix vesicles from chondrocytes and osteoblasts: their biogenesis, properties, functions and biomimetic models. Biochim. Biophys. Acta Gen. Subj., 2018, 1862: 532-546

[17] Wei Y et al. Extracellular vesicles derived from the mid-to-late stage of osteoblast differentiation markedly enhance osteogenesis in vitro and in vivo. Biochem. Biophys. Res. Commun., 2019, 514: 252-258

[18] Wang ZX et al. Aged bone matrix-derived extracellular vesicles as a messenger for calcification paradox. Nat. Commun., 2022, 13

[19] Li T et al. Matrix vesicles as a therapeutic target for vascular calcification. Front. Cell Dev. Biol., 2022, 10: 825622

[20] Wang Y et al. Bioinspired extracellular vesicles embedded with black phosphorus for molecular recognition-guided biomineralization. Nat. Commun., 2019, 10

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

[22] Man K et al. Epigenetic reprogramming enhances the therapeutic efficacy of osteoblast-derived extracellular vesicles to promote human bone marrow stem cell osteogenic differentiation. J. Extracell. Vesicles., 2021, 10: e12118

[23] Gerke V, Creutz CE, Moss SE. Annexins: Linking Ca2+ signalling to membrane dynamics. Nat. Rev. Mol. Cell Biol., 2005, 6: 449-461

[24] Xi Y, Ju R, Wang Y. Roles of Annexin A protein family in autophagy regulation and therapy. Biomed. Pharmacother., 2020, 130: 110591

[25] Pei T et al. Fluid shear stress regulates osteogenic differentiation via annexin A6-mediated autophagy in MC3T3-E1 cells. Int. J. Mol. Sci., 2022, 23: 15702

[26] Thouverey C et al. Proteomic characterization of biogenesis and functions of matrix vesicles released from mineralizing human osteoblast-like cells. J. Proteom., 2011, 74: 1123-1134

[27] Wang W, Xu J, Kirsch T. Annexin-mediated Ca2+ influx regulates growth plate chondrocyte maturation and apoptosis. J. Biol. Chem., 2003, 278: 3762-3769

[28] Rogers MA et al. Annexin A1-dependent tethering promotes extracellular vesicle aggregation revealed with single-extracellular vesicle analysis. Sci. Adv., 2020, 6: eabb1244

[29] Kim HJ, Kirsch T. Collagen/annexin V interactions regulate chondrocyte mineralization. J. Biol. Chem., 2008, 283: 10310-10317

[30] Moreau K et al. Transcriptional regulation of Annexin A2 promotes starvation-induced autophagy. Nat. Commun., 2015, 6

[31] Wang X et al. miR-214 targets ATF4 to inhibit bone formation. Nat. Med., 2013, 19: 93-100

[32] Chen CY et al. Extracellular vesicles from human urine-derived stem cells prevent osteoporosis by transferring CTHRC1 and OPG. Bone Res., 2019, 7: 18

[33] Buchet R et al. Isolation and characteristics of matrix vesicles. Methods Mol. Biol., 2013, 1053: 115-124

[34] Wuthier RE, Lipscomb GF. Matrix vesicles: structure, composition, formation and function in calcification. Front. Biosci. (Landmark Ed.), 2011, 16: 2812-2902

[35] Rilla K et al. Extracellular vesicles are integral and functional components of the extracellular matrix. Matrix Biol., 2019, 75–76: 201-219

[36] Al Halawani A et al. Extracellular vesicles: interplay with the extracellular matrix and modulated cell responses. Int. J. Mol. Sci., 2022, 23: 3389

[37] Bolean M et al. Matrix vesicle biomimetics harboring Annexin A5 and alkaline phosphatase bind to the native collagen matrix produced by mineralizing vascular smooth muscle cells. Biochim. Biophys. Acta Gen. Subj., 2020, 1864: 129629

[38] Roszkowska M et al. Collagen promotes matrix vesicle-mediated mineralization by vascular smooth muscle cells. J. Inorg. Biochem., 2018, 186: 1-9

[39] Zheng J et al. Extracellular vesicles degradation pathway based autophagy lysosome pathway. Am. J. Transl. Res., 2019, 11: 1170-1183

[40] Iwayama T et al. Osteoblastic lysosome plays a central role in mineralization. Sci. Adv., 2019, 5: eaax0672

[41] Chen X et al. Nrf2 epigenetic derepression induced by running exercise protects against osteoporosis. Bone Res., 2021, 9: 15

[42] Du X et al. The role of TGF-beta3 in cartilage development and osteoarthritis. Bone Res., 2023, 11: 2

[43] Zhou C et al. Hydrogel platform with tunable stiffness based on magnetic nanoparticles cross-linked GelMA for cartilage regeneration and its intrinsic biomechanism. Bioact. Mater., 2023, 25: 615-628

[44] Zhang et al Osteoporosis-decreased extracellular matrix stiffness impairs connexin 43-mediated gap junction intercellular communication in osteocytes. Acta Biochim. Biophys. Sin. (Shanghai), 2020, 52: 517-526

[45] Rutkovskiy A, Stenslokken KO, Vaage IJ. Osteoblast differentiation at a glance. Med. Sci. Monit. Basic. Res., 2016, 22: 95-106

[46] Yang Y et al. Transforming growth factor-β1-induced N-cadherin drives cell-cell communication through connexin43 in osteoblast lineage. Int. J. Oral. Sci., 2021, 13: 15

[47] Weitzmann MN, Ofotokun I. Physiological and pathophysiological bone turnover - role of the immune system. Nat. Rev. Endocrinol., 2016, 12: 518-532

[48] Liu Y et al. The role of mechano growth factor in chondrocytes and cartilage defects: a concise review. Acta Biochim. Biophys. Sin. (Shanghai), 2023, 55: 701-712

[49] Zhou C et al. Runx1 protects against the pathological progression of osteoarthritis. Bone Res, 2021, 9: 50

[50] Liu Y et al. Biomimetic fibers derived from an equidistant micropillar platform dictate osteocyte fate via mechanoreception. Nano Lett., 2023, 23: 7950-7960

[51] Chen H et al. Research progress on the role of extracellular vesicles derived from aging cells in osteoporosis. Biosci. Rep., 2023, 43: BSR20221775

[52] Zhang W et al. The role of extracellular vesicles in osteoporosis: a scoping review. Membr. (Basel), 2022, 12: 324

[53] Schmidt JR et al. Osteoblast-released matrix vesicles, regulation of activity and composition by sulfated and non-sulfated glycosaminoglycans. Mol. Cell Proteom., 2016, 15: 558-572

[54] Chaudhary SC et al. Phosphate induces formation of matrix vesicles during odontoblast-initiated mineralization in vitro. Matrix Biol., 2016, 52-54: 284-300

[55] Kapustin AN et al. Vascular smooth muscle cell calcification is mediated by regulated exosome secretion. Circ. Res., 2015, 116: 1312-1323

[56] Bakhshian Nik A, Hutcheson JD, Aikawa E. Extracellular vesicles as mediators of cardiovascular calcification. Front. Cardiovasc. Med., 2017, 4: 78

[57] Sariano PA et al. Convection and extracellular matrix binding control interstitial transport of extracellular vesicles. J. Extracell. Vesicles, 2023, 12: e12323

[58] Itel F, Skovhus Thomsen J, Städler B. Matrix vesicles-containing microreactors as support for bonelike osteoblasts to enhance biomineralization. ACS Appl. Mater. Interfaces, 2018, 10: 30180-30190

[59] Pepe J et al. Characterization of extracellular vesicles in osteoporotic patients compared to osteopenic and healthy controls. J. Bone Miner. Res., 2022, 37: 2186-2200

[60] van Niel G, D’Angelo G, Raposo G. Shedding light on the cell biology of extracellular vesicles. Nat. Rev. Mol. Cell Bio., 2018, 19: 213-228

[61] Gerke V, Moss SE. Annexins: from structure to function. Physiol. Rev., 2002, 82: 331-371

[62] Grewal T et al. Annexins in adipose tissue: novel players in obesity. Int. J. Mol. Sci., 2019, 20: 3449

[63] Balcerzak M et al. Proteome analysis of matrix vesicles isolated from femurs of chicken embryo. Proteomics, 2008, 8: 192-205

[64] Genetos DC et al. Impaired osteoblast differentiation in annexin A2- and -A5-deficient cells. PLoS One, 2014, 9: e107482

[65] Brachvogel B et al. Sequential expression of annexin A5 in the vasculature and skeletal elements during mouse development. Mech. Dev., 2001, 109: 389-393

[66] Haut Donahue TL et al. Annexin V disruption impairs mechanically induced calcium signaling in osteoblastic cells. Bone, 2004, 35: 656-663

[67] Lenzini S et al. Matrix mechanics and water permeation regulate extracellular vesicle transport. Nat. Nanotechnol., 2020, 15: 217-223

[68] Ma Q et al. Osteoclast-derived apoptotic bodies couple bone resorption and formation in bone remodeling. Bone Res., 2021, 9: 5

[69] Landis WJ et al. An overview of vertebrate mineralization with emphasis on collagen-mineral interaction. Gravit. Space Biol. Bull., 1999, 12: 15-26

[70] Hasegawa T. Ultrastructure and biological function of matrix vesicles in bone mineralization. Histochem. Cell Biol., 2018, 149: 289-304

[71] Kirsch T et al. The roles of annexins and types II and X collagen in matrix vesicle-mediated mineralization of growth plate cartilage. J. Biol. Chem., 2000, 275: 35577-35583

[72] Zhang C et al. Coupling of integrin alpha5 to annexin A2 by flow drives endothelial activation. Circ. Res., 2020, 127: 1074-1090

[73] Blaser MC, Aikawa E. Roles and regulation of extracellular vesicles in cardiovascular mineral metabolism. Front. Cardiovasc. Med., 2018, 5: 187

[74] Behera J et al. Mechanisms of autophagy and mitophagy in skeletal development, diseases and therapeutics. Life Sci., 2022, 301: 120595

[75] Wang, J. et al. The role of autophagy in bone metabolism and clinical significance. Autophagy 1-19 (2023).

[76] Zhang HT et al. TRIM21-regulated Annexin A2 plasma membrane trafficking facilitates osteosarcoma cell differentiation through the TFEB-mediated autophagy. Cell Death. Dis., 2021, 12

[77] Ji T, Kohane DS. Nanoscale systems for local drug delivery. Nano Today, 2019, 28: 100765

[78] Zhang H et al. Bone/cartilage targeted hydrogel: Strategies and applications. Bioact. Mater., 2022, 23: 156-169

[79] Rotman SG et al. Drug delivery systems functionalized with bone mineral seeking agents for bone targeted therapeutics. J. Control. Release, 2018, 269: 88-99

[80] Kang TH et al. Annexin A5 as an immune checkpoint inhibitor and tumor-homing molecule for cancer treatment. Nat. Commun., 2020, 11

[81] Haeckel A et al. Long-circulating XTEN864-annexin A5 fusion protein for phosphatidylserine-related therapeutic applications. Apoptosis, 2021, 26: 534-547

[82] Zhang X et al. Phosphatidylserine externalized on the colonic capillaries as a novel pharmacological target for IBD therapy. Signal Transduct. Target Ther., 2021, 6: 235

[83] Zhu H et al. A protocol for isolation and culture of mesenchymal stem cells from mouse compact bone. Nat. Protoc., 2010, 5: 550-560

[84] Liu B et al. A protocol for isolation and identification and comparative characterization of primary osteoblasts from mouse and rat calvaria. Cell Tissue Bank, 2019, 20: 173-182

[85] Feng T et al. Fluid shear stress-induced exosomes from liver cancer cells promote activation of cancer-associated fibroblasts via IGF2-PI3K axis. Front. Biosci. (Landmark Ed.), 2022, 27: 104

[86] Yu H et al. Fluid shear stress activates YAP to promote epithelial-mesenchymal transition in hepatocellular carcinoma. Mol. Oncol., 2021, 15: 3164-3183

Funding

National Natural Science Foundation of China (National Science Foundation of China)(31971239)