Activation of Wnt signaling by mechanical loading is impaired in the bone of old mice

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Holguin, Nilsson¹; Brodt, Michael D.¹; Silva, Matthew J.^1,2

Activation of Wnt signaling by mechanical loading is impaired in the bone of old mice

J Bone Miner Res. December 2016;31(12):2215-2226

©2016 American Society for Bone and Mineral Research

Affiliations

¹Department of Orthopaedic Surgery, Musculoskeletal Research Center, Washington University, St. Louis, MO, USA

²Department of Biomedical Engineering, Washington University, St. Louis, MO, USA
Abstract and keywords

Aging diminishes bone formation engendered by mechanical loads, but the mechanism for this impairment remains unclear. Because Wnt signaling is required for optimal loading-induced bone formation, we hypothesized that aging impairs the load-induced activation of Wnt signaling. We analyzed dynamic histomorphometry of 5-month-old, 12-month-old, and 22-month-old C57Bl/6JN mice subjected to multiple days of tibial compression and corroborated an age-related decline in the periosteal loading response on day 5. Similarly, 1 day of loading increased periosteal and endocortical bone formation in young-adult (5-month-old) mice, but old (22-month-old) mice were unresponsive. These findings corroborated mRNA expression of genes related to bone formation and the Wnt pathway in tibias after loading. Multiple bouts (3 to 5 days) of loading upregulated bone formation–related genes, e.g., Osx and Col1a1, but older mice were significantly less responsive. Expression of Wnt negative regulators, Sost and Dkk1, was suppressed with a single day of loading in all mice, but suppression was sustained only in young-adult mice. Moreover, multiple days of loading repeatedly suppressed Sost and Dkk1 in young-adult, but not in old tibias. The age-dependent response to loading was further assessed by osteocyte staining for Sclerostin and LacZ in tibia of TOPGAL mice. After 1 day of loading, fewer osteocytes were Sclerostin-positive and, corroboratively, more osteocytes were LacZ-positive (Wnt active) in both 5-month-old and 12-month-old mice. However, although these changes were sustained after multiple days of loading in 5-month-old mice, they were not sustained in 12-month-old mice. Last, Wnt1 and Wnt7b were the most load-responsive of the 19 Wnt ligands. However, 4 hours after a single bout of loading, although their expression was upregulated threefold to 10-fold in young-adult mice, it was not altered in old mice. In conclusion, the reduced bone formation response of aged mice to loading may be due to failure to sustain Wnt activity with repeated loading.

Keywords: AGING; WNT/β-CATENIN SIGNALING; ANABOLIC; OSTEOCYTE; OSTEOBLAST
Links

DOI: 10.1002/jbmr.2900

PubMed: 27357062

PubMedCentral: PMC5397287

WoS: 000389445100018
History

Received: 2016-04-20

Revised: 2016-06-24

Accepted: 2016-06-27

Online: 2016-06-30

Cited Works (23)

Year	Entry
2011	Lynch ME, Main RP, Xu Q, Schmicker TL, Schaffler MB, Wright TM, van der Meulen MCH. Tibial compression is anabolic in the adult mouse skeleton despite reduced responsiveness with aging. Bone. September 2011;49(3):439-446.
2002	Boyden LM, Mao J, Belsky J, Mitzner L, Farhi A, Mitnick MA, Wu D, Insogna K, Lifton RP. High bone density due to a mutation in LDL-receptor–related protein 5. NEJM. May 16, 2002;346(20):1513-1521.
2006	Robinson JA, Chatterjee-Kishore M, Yaworsky PJ, Cullen DM, Zhao W, Li C, Kharode Y, Sauter L, Babij P, Brown EL, Hill AA, Akhter MP, Johnson ML, Recker RR, Komm BS, Bex FJ. Wnt/β-catenin signaling is a normal physiological response to mechanical loading in bone. J Biol Chem. October 20, 2006;281(42):31720-31728.
1992	Rubin CT, Bain SD, McLeod KJ. Suppression of the osteogenic response in the aging skeleton. Calcif Tiss Int. April 1992;50(4):306-313.
2014	Meakin LB, Galea GL, Sugiyama T, Lanyon LE, Price JS. Age-related impairment of bones' adaptive response to loading in mice is associated with sex-related deficiencies in osteoblasts but no change in osteocytes. J Bone Miner Res. August 2014;29(8):1859-1871.
2012	Silva MJ, Brodt MD, Lynch MA, Stephens AL, Wood DJ, Civitelli R. Tibial loading increases osteogenic gene expression and cortical bone volume in mature and middle-aged mice. PLoS One. April 2012;7(4):e34980.
2012	Tu X, Rhee Y, Condon KW, Bivi N, Allen MR, Dwyer D, Stolina M, Turner CH, Robling AG, Plotkin LI, Bellido T. Sost downregulation and local Wnt signaling are required for the osteogenic response to mechanical loading. Bone. January 2012;50(1):209-217.
1995	Turner CH, Takano Y, Owan I. Aging changes mechanical loading thresholds for bone formation in rats. J Bone Miner Res. October 1995;10(10):1544-1549.
2014	Holguin N, Brodt MD, Sanchez ME, Silva MJ. Aging diminishes lamellar and woven bone formation induced by tibial compression in adult C57BL/6. Bone. August 2014;65:83-91.
2001	Gong Y, Slee RB, Fukai N, Rawadi G, Roman-Roman S, Reginato AM, Wang H, Cundy T, Glorieux FH, Lev D, Zacharin M, Oexle K, Marcelino J, Suwairi W, Heeger S, Sabatakos G, Apte S, Adkins WN, Allgrove J, Arslan-Kirchner M, Batch JA, Beighton P, Black GCM, Boles RG, Boon LM, Borrone C, Brunner HG, Carle GF, Dallapiccola B, De Paepe A, Floege B, Halfhide ML, Hall B, Hennekam RC, Hirose T, Jans A, Jüppner H, Kim CA, Keppler-Noreuil K, Kohlschuetter A, LaCombe D, Lambert M, Lemyre E, Letteboer T, Peltonen L, Ramesar RS, Romanengo M, Somer H, Steichen-Gersdorf E, Steinmann B, Sullivan B, Superti-Furga A, Swoboda W, van den Boogaard M-J, Van Hul W, Vikkula M, Votruba M, Zabel B, Garcia T, Baron R, Olsen BR, Warman ML. LDL receptor-related protein 5 (LRP5) affects bone accrual and eye development. Cell. November 16, 2001;107(4):513-523.
2010	Brodt MD, Silva MJ. Aged mice have enhanced endocortical response and normal periosteal response compared with young-adult mice following 1 week of axial tibial compression. J Bone Miner Res. September 2010;25(9):2006-2015.
2008	Bonewald LF, Johnson ML. Osteocytes, mechanosensing and Wnt signaling. Bone. April 2008;42(4):606-615.
2013	Baron R, Kneissel M. WNT signaling in bone homeostasis and disease: from human mutations to treatments. Nat Med. February 2013;19(2):179-192.
2014	Patel TK, Brodt MD, Silva MJ. Experimental and finite element analysis of strains induced by axial tibial compression in young-adult and old female C57Bl/6 mice. J Biomech. January 22, 2014;47(2):451-457.
2012	Sugiyama T, Meakin LB, Browne WJ, Galea GL, Price JS, Lanyon LE. Bones' adaptive response to mechanical loading is essentially linear between the low strains associated with disuse and the high strains associated with the lamellar/woven bone transition. J Bone Miner Res. August 2012;27(8):1784-1793.
2011	Mantila Roosa SM, Liu Y, Turner CH. Gene expression patterns in bone following mechanical loading. J Bone Miner Res. January 2011;26(1):100-112.
2003	Srinivasan S, Agans SC, King KA, Moy NY, Poliachik SL, Gross TS. Enabling bone formation in the aged skeleton via rest-inserted mechanical loading. Bone. December 2003;33(6):946-955.
2012	Moustafa A, Sugiyama T, Prasad J, Zaman G, Gross TS, Lanyon LE, Price JS. Mechanical loading-related changes in osteocyte sclerostin expression in mice are more closely associated with the subsequent osteogenic response than the peak strains engendered. Osteoporos Int. April 2012;23(4):1225-1234.
2003	Ferguson VL, Ayers RA, Bateman TA, Simske SJ. Bone development and age-related bone loss in male C57BL/6J mice. Bone. March 2003;32(3):387-398.
2013	Dempster DW, Compston JE, Drezner MK, Glorieux FH, Kanis JA, Malluche H, Meunier PJ, Ott SM, Recker RR, Parfitt AM. Standardized nomenclature, symbols, and units for bone histomorphometry: a 2012 update of the report of the ASBMR Histomorphometry Nomenclature Committee. J Bone Miner Res. January 2013;28(1):2-17.
1978	Lips P, Courpron P, Meunier PJ. Mean wall thickness of trabecular bone packets in the human iliac crest: changes with age. Calcif Tiss Res. 1978;26(1):13-17.
1994	Raab-Cullen DM, Thiede MA, Petersen DN, Kimmel DB, Recker RR. Mechanical loading stimulates rapid changes in periosteal gene expression. Calcif Tiss Int. December 1994;55(6):473-478.
2008	Robling AG, Niziolek PJ, Baldridge LA, Condon KW, Allen MR, Alam I, Mantila SM, Gluhak-Heinrich J, Bellido TM, Harris SE, Turner CH. Mechanical stimulation of bone in vivo reduces osteocyte expression of Sost/sclerostin. J Biol Chem. February 29, 2008;283(9):5866-5875.

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2020	Morse A, Ko FC, McDonald MM, Lee LR, Schindeler A, van der Meulen MCH, Little DG. Increased anabolic bone response in Dkk1 KO mice following tibial compressive loading. Bone. February 2020;131:115054.
2021	Coates BA, McKenzie JA, Yoneda S, Silva MJ. Interleukin-6 (IL-6) deficiency enhances intramembranous osteogenesis following stress fracture in mice. Bone. February 2021;143:115737.
2021	Harris TL, Silva MJ. Gene expression of intracortical bone demonstrates loading-induced increases in Wnt1 and Ngf and inhibition of bone remodeling processes. Bone. September 2021;150:116019.
2022	Abdurahman A, Li X, Li J, Liu D, Zhai L, Wang X, Zhang Y, Meng Y, Yokota H, Zhang P. Loading-driven PI3K/Akt signaling and erythropoiesis enhanced angiogenesis and osteogenesis in a postmenopausal osteoporosis mouse model. Bone. April 2022;157:116346.
2022	Rooney AM, Ayobami OO, Kelly NH, Schimenti JC, Ross FP, van der Meulen MCH. Bone mass and adaptation to mechanical loading are sexually dimorphic in adult osteoblast-specific ERα knockout mice. Bone. May 2022;158:116349.
2020	Carina V, Della Bella E, Costa V, Bellavia D, Veronesi F, Cepollaro S, Fini M, Giavaresi G. Bone's response to mechanical loading in aging and osteoporosis: molecular mechanisms. Calcif Tiss Int. October 2020;107(4):301-318.
2020	Chermside-Scabbo CJ, Harris TL, Brodt MD, Braenne I, Zhang B, Farber CR, Silva MJ. Old mice have less transcriptional activation but similar periosteal cell proliferation compared to young‐adult mice in response to in vivo mechanical loading. J Bone Miner Res. September 2020;35(9):1751-1764.
2022	Lawson LY, Brodt MD, Migotsky N, Chermside-Scabbo CJ, Palaniappan R, Silva MJ. Osteoblast-specific wnt secretion is required for skeletal homeostasis and loading-induced bone formation in adult mice. J Bone Miner Res. January 2022;37(1):108-120.
2022	McGregor NE, Walker EC, Chan ASM, Poulton IJ, Cho EH-J, Windahl SH, Sims NA. STAT3 hyperactivation due to SOCS3 deletion in murine osteocytes accentuates responses to exercise- and load-induced bone formation. J Bone Miner Res. March 2022;37(3):547-558.
2022	Kroon T, Bhadouria N, Niziolek P, Edwards D III, Choi R, Clinkenbeard EL, Robling A, Holguin N. Suppression of sost/sclerostin and dickkopf-1 augment intervertebral disc structure in mice. J Bone Miner Res. June 2022;37(6):1156-1169.
2022	Chlebek C, Moore JA, Ross FP, van der Meulen MCH. Molecular identification of spatially distinct anabolic responses to mechanical loading in murine cortical bone. J Bone Miner Res. November 2022;37(11):2277-2287.
2018	Sun D, Brodt MD, Zannit HM, Holguin N, Silva MJ. Evaluation of loading parameters for murine axial tibial loading: stimulating cortical bone formation while reducing loading duration. J Orthop Res. February 2018;36(2):682-691.
2022	Ko FC, Moran MM, Ross RD, Sumner DR. Activation of canonical Wnt signaling accelerates intramembranous bone regeneration in male mice. J Orthop Res. August 2022;40(8):1834-1843.
2021	Wilmoth RL. Development of a 3D Ex Vivo Culture System to Study Osteocyte Mechanobiology [PhD thesis]. University of Colorado; 2021.
2019	Campi AE. Mechanosensitive Ca²⁺ Signaling of Ex Vivo Osteocytes in Aging and Treatment [PhD thesis]. Columbia University; 2019.
2020	Rooney A. Interactions Among Estrogen Receptor-Alpha, Parathyroid Hormone, and Mechanical Loading in Skeletal Health [PhD thesis]. Ithaca, NY: Cornell University; December 2020.
2022	Chlebek C. Identification of Novel Cellular Pathways Involved in Bone Mechanotransduction Using Transcriptomics [PhD thesis]. Ithaca, NY: Cornell University; May 2022.
2019	Piet J. Restoring Mechano-Adaptation in Aged Murine Bone [PhD thesis]. Northeastern University; March 2019.
2019	Podlisny DR. Effects of Poroelasticity on Mechanoadaptation in Bone [Master's thesis]. Northeastern University; May 2019.
2017	Morse AR. The Role of Wnt/β-Catenin Antagonists - Sclerostin and DKK1 - in Bone Homeostasis and Mechanotransduction [PhD thesis]. University of Sydney; 2017.
2019	Mazur C. Osteocyte Regulation of Bone Quality in Homeostasis and in Skeletal Disease [PhD thesis]. San Francisco, University of California; 2019.
2020	Pei S. Osteocyte Mechanosensing: Interstitial Fluid Flow, Cellular Response, and Turnover of Pericellular Matrix [PhD thesis]. University of Delaware; 2020.
2020	Coates BA. The Role of Gene Transcription and Inflammatory Cytokines in Bone Fracture Repair [PhD thesis]. St. Louis, MO: Washington University in St. Louis; May 2020.
2021	Harris TL. Mechanisms for Osteoblast and Osteocyte Initiation and Sustainment of Bone Formation in Young-Adult and Aged Mice [PhD thesis]. St. Louis, MO: Washington University in St. Louis; August 2021.