PDGFRα reporter activity identifies periosteal progenitor cells critical for bone formation and fracture repair

Abstract

The outer coverings of the skeleton, which is also known as the periosteum, are arranged in concentric layers and act as a reservoir for tissue-specific bone progenitors. The cellular heterogeneity within this tissue depot is being increasingly recognized. Here, inducible PDGFRα reporter animals were found to mark a population of cells within the periosteum that act as a stem cell reservoir for periosteal appositional bone formation and fracture repair. During these processes, PDGFRα reporter⁺ progenitors give rise to Nestin⁺ periosteal cells before becoming osteoblasts and osteocytes. The diphtheria toxin-mediated ablation of PDGFRα reporter⁺ cells led to deficits in cortical bone formation during homeostasis and a diminutive hard callus during fracture repair. After ossicle transplantation, both mouse PDGFRα reporter⁺ periosteal cells and human Pdgfrα⁺ periosteal progenitors expand, ossify, and recruit marrow to a greater extent than their counterpart periosteal cells, whereas PDGFRα reporter⁻ periosteal cells exhibit a predisposition to chondrogenesis in vitro. Total RNA sequencing identified enrichment of the secreted factors Fermt3 and Ptpn6 within PDGFRα reporter⁺ periosteal cells, which partly underlie the osteoblastogenic features of this cell population.

References

[1] Allen MR, Hock JM, Burr DB. Periosteum: biology, regulation, and response to osteoporosis therapies. Bone, 2004, 35: 1003-1012

[2] Seeman E. Periosteal bone formation-a neglected determinant of bone strength. N. Engl. J. Med., 2003, 349: 320-323

[3] Jones DB, Nolte H, Scholubbers JG, Turner E, Veltel D. Biochemical signal transduction of mechanical strain in osteoblast-like cells. Biomaterials, 1991, 12: 101-110

[4] Chyun YS, Kream BE, Raisz LG. Cortisol decreases bone formation by inhibiting periosteal cell proliferation. Endocrinology, 1984, 114: 477-480

[5] Ono N et al. Vasculature-associated cells expressing nestin in developing bones encompass early cells in the osteoblast and endothelial lineage. Dev. Cell, 2014, 29: 330-339

[6] Gao B et al. Macrophage-lineage TRAP+ cells recruit periosteum-derived cells for periosteal osteogenesis and regeneration. J. Clin. Invest., 2019, 129: 2578-2594

[7] Shi Y et al. Gli1 identifies osteogenic progenitors for bone formation and fracture repair. Nat. Commun., 2017, 8

[8] Ortinau LC et al. Identification of functionally distinct Mx1+alphaSMA+ periosteal skeletal stem cells. Cell Stem Cell, 2019, 25: 784-796.e785

[9] Debnath S et al. Discovery of a periosteal stem cell mediating intramembranous bone formation. Nature, 2018, 562: 133-139

[10] Chan CKF et al. Identification of the human skeletal stem cell. Cell, 2018, 175: 43-56 e21

[11] Bohm AM et al. Activation of skeletal stem and progenitor cells for bone regeneration is driven by PDGFRbeta signaling. Dev. Cell, 2019, 51: 236-254.e212

[12] Houlihan DD et al. Isolation of mouse mesenchymal stem cells on the basis of expression of Sca-1 and PDGFR-alpha. Nat. Protoc., 2012, 7: 2103-2111

[13] Arrighi N et al. Characterization of adipocytes derived from fibro/adipogenic progenitors resident in human skeletal muscle. Cell Death Dis., 2015, 6

[14] Wang Y et al. PDGFRalpha marks distinct perivascular populations with different osteogenic potential within adipose tissue. Stem Cells, 2020, 38: 276-290

[15] Cherry, H. M., Roelofs, A. J., Kurth, T. B. & De Bari, C. In vivo phenotypic characterisation of nucleoside label-retaining cells in mouse periosteum. Eur. Cell Mater. 27, 185–195, discussion 195 (2014).

[16] Morikawa S et al. Prospective identification, isolation, and systemic transplantation of multipotent mesenchymal stem cells in murine bone marrow. J. Exp. Med, 2009, 206: 2483-2496

[17] Noseda M et al. PDGFRalpha demarcates the cardiogenic clonogenic Sca1⁺ stem/progenitor cell in adult murine myocardium. Nat. Commun., 2015, 6

[18] Farahani RM, Xaymardan M. Platelet-derived growth factor receptor alpha as a marker of mesenchymal stem cells in development and stem cell biology. Stem Cells Int., 2015, 2015: 362753

[19] Mitlak BH et al. The effect of systemically administered PDGF-BB on the rodent skeleton. J. Bone Min. Res., 1996, 11: 238-247

[20] Wang X et al. PDGF modulates BMP2-induced osteogenesis in periosteal progenitor cells. JBMR Plus, 2019, 3: e10127

[21] Augustin G, Antabak A, Davila S. The periosteum. Part 1: anatomy, histology and molecular biology. Injury, 2007, 38: 1115-1130

[22] Qi C, Xiaofeng X, Xiaoguang W. Effects of toll-like receptors 3 and 4 in the osteogenesis of stem cells. Stem Cells Int., 2014, 2014: 917168

[23] Lee CH, Huang YL, Liao JF, Chiou WF. Ugonin K-stimulated osteogenesis involves estrogen receptor-dependent activation of non-classical Src signaling pathway and classical pathway. Eur. J. Pharm., 2012, 676: 26-33

[24] Kristianto J, Johnson MG, Afzal R, Blank RD. Endothelin signaling in bone. Endocrinol. Metab. Clin. North Am., 2017, 46: 51-62

[25] Saelao P et al. Novel insights into the host immune response of chicken Harderian gland tissue during Newcastle disease virus infection and heat treatment. BMC Vet. Res., 2018, 14: 280

[26] Yao Y et al. Global profiling of the gene expression and alternative splicing events during hypoxia-regulated chondrogenic differentiation in human cartilage endplate-derived stem cells. Genomics, 2016, 107: 170-177

[27] Kita K, Kimura T, Nakamura N, Yoshikawa H, Nakano T. PI3K/Akt signaling as a key regulatory pathway for chondrocyte terminal differentiation. Genes Cells, 2008, 13: 839-850

[28] Guo W, Fan Z, Wang S, Du J. ALK5 is essential for tooth germ differentiation during tooth development. Biotech. Histochem., 2019, 94: 481-490

[29] Wang S et al. Insulin-like growth factor 1 can promote the osteogenic differentiation and osteogenesis of stem cells from apical papilla. Stem Cell Res., 2012, 8: 346-356

[30] Chang J et al. Noncanonical Wnt-4 signaling enhances bone regeneration of mesenchymal stem cells in craniofacial defects through activation of p38 MAPK. J. Biol. Chem., 2007, 282: 30938-30948

[31] Shen J et al. Effects of WNT3A and WNT16 on the osteogenic and adipogenic differentiation of perivascular stem/stromal cells. Tissue Eng. Part A, 2018, 24: 68-80

[32] Dranse HJ, Sampaio AV, Petkovich M, Underhill TM. Genetic deletion of Cyp26b1 negatively impacts limb skeletogenesis by inhibiting chondrogenesis. J. Cell Sci., 2011, 124: 2723-2734

[33] Delprat B et al. Downregulation of otospiralin, a novel inner ear protein, causes hair cell degeneration and deafness. J. Neurosci., 2002, 22: 1718-1725

[34] Tamamura Y, Katsube K, Mera H, Itokazu M, Wakitani S. Irx3 and Bmp2 regulate mouse mesenchymal cell chondrogenic differentiation in both a Sox9-dependent and -independent manner. J. Cell Physiol., 2017, 232: 3317-3336

[35] Islam A et al. In vitro chondrogenic potency of surplus chondrocytes from autologous transplantation procedures does not predict short-term clinical outcomes. BMC Musculoskelet. Disord., 2019, 20

[36] Bao Q et al. alphaB-crystallin (CRYAB) regulates the proliferation, apoptosis, synthesis and degradation of extracellular matrix of chondrocytes in osteoarthritis. Exp. Cell Res., 2019, 382: 111459

[37] Schmidt S et al. Kindlin-3-mediated signaling from multiple integrin classes is required for osteoclast-mediated bone resorption. J. Cell Biol., 2011, 192: 883-897

[38] Kerr BA et al. Kindlin-3 Mutation in Mesenchymal Stem Cells Results in Enhanced Chondrogenesis. Exp. Cell Res., 2021, 399: 112456

[39] Jiang M et al. SHP1 regulates bone mass by directing mesenchymal stem cell differentiation. Cell Rep., 2016, 16: 769-780

[40] Murakami J et al. Vascular endothelial growth factor-C induces osteogenic differentiation of human mesenchymal stem cells through the ERK and RUNX2 pathway. Biochem. Biophys. Res. Commun., 2017, 484: 710-718

[41] Duchamp de Lageneste O et al. Periosteum contains skeletal stem cells with high bone regenerative potential controlled by Periostin. Nat. Commun., 2018, 9

[42] Leucht P et al. Embryonic origin and Hox status determine progenitor cell fate during adult bone regeneration. Development, 2008, 135: 2845-2854

[43] Chan CK et al. Identification and specification of the mouse skeletal stem cell. Cell, 2015, 160: 285-298

[44] He X et al. Sox9 positive periosteal cells in fracture repair of the adult mammalian long bone. Bone, 2017, 103: 12-19

[45] Li A et al. PDGF-AA promotes osteogenic differentiation and migration of mesenchymal stem cell by down-regulating PDGFRalpha and derepressing BMP-Smad1/5/8 signaling. PLoS One, 2014, 9: e113785

[46] Zhang M et al. The effects of platelet-derived growth factor-BB on bone marrow stromal cell-mediated vascularized bone regeneration. Stem Cells Int., 2018, 2018: 3272098

[47] Caplan AI, Correa D. PDGF in bone formation and regeneration: new insights into a novel mechanism involving MSCs. J. Orthop. Res., 2011, 29: 1795-1803

[48] Doherty L et al. A PDGFRbeta-PI3K signaling axis mediates periosteal cell activation during fracture healing. PLoS One, 2019, 14: e0223846

[49] Yakar S, Werner H, Rosen CJ. Insulin-like growth factors: actions on the skeleton. J. Mol. Endocrinol., 2018, 61: T115-T137

[50] Moverare-Skrtic S et al. Osteoblast-derived WNT16 represses osteoclastogenesis and prevents cortical bone fragility fractures. Nat. Med., 2014, 20: 1279-1288

[51] Anais J et al. Direct contribution of skeletal muscle mesenchymal progenitors to bone repair. Nat. Commun., 2021, 12: 2860

[52] Kurenkova AD, Medvedeva EV, Newton PT, Chagin AS. Niches for skeletal stem cells of mesenchymal origin. Front. Cell Dev. Biol., 2020, 8: 592

[53] Thuillier R et al. Interdependence of platelet-derived growth factor and estrogen-signaling pathways in inducing neonatal rat testicular gonocytes proliferation. Biol. Reprod., 2010, 82: 825-836

[54] Ambhore NS et al. Estrogen receptor beta signaling inhibits PDGF induced human airway smooth muscle proliferation. Mol. Cell Endocrinol., 2018, 476: 37-47

[55] Finlay GA, Hunter DS, Walker CL, Paulson KE, Fanburg BL. Regulation of PDGF production and ERK activation by estrogen is associated with TSC2 gene expression. Am. J. Physiol. Cell Physiol., 2003, 285: C409-C418

[56] Kang SH, Fukaya M, Yang JK, Rothstein JD, Bergles DE. NG2+ CNS glial progenitors remain committed to the oligodendrocyte lineage in postnatal life and following neurodegeneration. Neuron, 2010, 68: 668-681

[57] Kramann R et al. Perivascular Gli1+ progenitors are key contributors to injury-induced organ fibrosis. Cell Stem Cell, 2015, 16: 51-66

[58] Li Z et al. Fracture repair requires TrkA signaling by skeletal sensory nerves. J. Clin. Invest., 2019, 129: 5137-5150

Funding

U.S. Department of Health & Human Services | NIH | National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)(R01 AR070773)

U.S. Department of Health & Human Services | NIH | National Institute of Dental and Craniofacial Research (NIDCR)(R21 DE027922)

U.S. Department of Defense (United States Department of Defense)(W81XWH-18-1-0121)

American Cancer Society (American Cancer Society, Inc.)(Research Scholar Grant, RSG-18-027-01-CSM)

Maryland Stem Cell Research Fund (MSCRF)

MTF Biologics