Primary Cilia‐Mediated Mechanotransduction in Human Mesenchymal Stem Cells

Abstract

Physical loading is a potent stimulus required to maintain bone homeostasis, partly through the renewal and osteogenic differentiation of mesenchymal stem cells (MSCs). However, the mechanism by which MSCs sense a biophysical force and translate that into a biochemical bone forming response (mechanotransduction) remains poorly understood. The primary cilium is a single sensory cellular extension, which has recently been shown to demonstrate a role in cellular mechanotransduction and MSC lineage commitment. In this study, we present evidence that short periods of mechanical stimulation in the form of oscillatory fluid flow (OFF) is sufficient to enhance osteogenic gene expression and proliferation of human MSCs (hMSCs). Furthermore, we demonstrate that the cilium mediates fluid flow mechanotransduction in hMSCs by maintaining OFF-induced increases in osteogenic gene expression and, surprisingly, to limit OFF-induced increases in proliferation. These data therefore demonstrate a pro-osteogenic mechanosensory role for the primary cilium, establishing a novel mechanotransduction mechanism in hMSCs. Based on these findings, the application of OFF may be a beneficial component of bioreactor-based strategies to form bone-like tissues suitable for regenerative medicine and also highlights the cilium as a potential therapeutic target for efforts to mimic loading with the aim of preventing bone loss during diseases such as osteoporosis. Furthermore, this study demonstrates a role for the cilium in controlling mechanically mediated increases in the proliferation of hMSCs, which parallels proposed models of polycystic kidney disease. Unraveling the mechanisms leading to rapid proliferation of mechanically stimulated MSCs with defective cilia could provide significant insights regarding ciliopathies and cystic diseases.

Shown for reading. Its publisher allows quoting it, not republishing it.

References 73

1. 1.

   Corral DA, Amling M, Priemel M, et al. Dissociation between bone resorption and bone formation in osteopenic transgenic mice. Proc Natl Acad Sci USA. 1998;95:13835-13840.

   Not in our library10.1073/pnas.95.23.13835

2. 2.

   Gurkan UA, Akkus O. The mechanical environment of bone marrow: A review. Ann Biomed Eng. 2008;36:1978-1991.

   Not in our library10.1007/s10439-008-9577-x

3. 3.

   Turner CH, Owan I, Alvey T, et al. Recruitment and proliferative responses of osteoblasts after mechanical loading in vivo determined using sustained-release bromodeoxyuridine. Bone. 1998;22:463-469.

   Not in our library10.1016/s8756-3282(98)00041-6

4. 4.

   Rubin CT, Capilla E, Luu YK, et al. Adipogenesis is inhibited by brief, daily exposure to high-frequency, extremely low-magnitude mechanical signals. Proc Natl Acad Sci USA. 2007;104:17879-17984.

   Not in our library10.1073/pnas.0708467104

5. 5.

   David V, Martin A, Lafage-Proust MH, et al. Mechanical loading down-regulates peroxisome proliferator-activated receptor gamma in bone marrow stromal cells and favors osteoblastogenesis at the expense of adipogenesis. Endocrinology. 2007;148:2553-2562.

   Not in our library10.1210/en.2006-1704

6. 6.

   Barou O, Palle S, Vico L, et al. Hindlimb unloading in rat decreases preosteoblast proliferation assessed in vivo with BrdU incorporation. Am J Physiol. 1998;274(1 pt 1):E108-E114.

   Not in our library10.1152/ajpendo.1998.274.1.e108

7. 7.

   Wronski TJ, Morey-Holton ER, Doty SB, et al. Histomorphometric analysis of rat skeleton following spaceflight. Am J Physiol. 1987;252(2 pt 2):R252-R255.

   Not in our library10.1152/ajpregu.1987.252.2.r252

8. 8.
   Mechanotherapy: how physical therapists’ prescription of exercise promotes tissue repair · 2009

   No notice
9. 9.

   McCoy RJ, O'Brien FJ. Influence of shear stress in perfusion bioreactor cultures for the development of three-dimensional bone tissue constructs: A review. Tissue Eng Part B, Rev. 2010;16:587-601.

   Not in our library10.1089/ten.teb.2010.0370

10. 10.

    Potier E, Noailly J, Ito K. Directing bone marrow-derived stromal cell function with mechanics. J Biomech. 2010;43:807-817.

    Not in our library10.1016/j.jbiomech.2009.11.019

11. 11.

    Castillo AB, Jacobs CR. Mesenchymal stem cell mechanobiology. Curr Osteoporosis Rep. 2010;8:98-104.

    Not in our library10.1007/s11914-010-0015-2

12. 12.

    Kelly DJ, Jacobs CR. The role of mechanical signals in regulating chondrogenesis and osteogenesis of mesenchymal stem cells. Birth Defects Res C Embryo Today. 2010;90:75-85.

    Not in our library10.1002/bdrc.20173

13. 13.

    Kreke MR, Huckle WR, Goldstein AS. Fluid flow stimulates expression of osteopontin and bone sialoprotein by bone marrow stromal cells in a temporally dependent manner. Bone. 2005;36:1047-1055.

    Not in our library10.1016/j.bone.2005.03.008

14. 14.

    Bancroft GN, Sikavitsas VI, van den DolderJ, et al. Fluid flow increases mineralized matrix deposition in 3D perfusion culture of marrow stromal osteoblasts in a dose-dependent manner. Proc Natl Acad Sci USA. 2002;99:12600-12605.

    Not in our library10.1073/pnas.202296599

15. 15.

    McGarry JG, Klein-Nulend J, Prendergast PJ. The effect of cytoskeletal disruption on pulsatile fluid flow-induced nitric oxide and prostaglandin E2 release in osteocytes and osteoblasts. Biochem Biophys Res Commun. 2005;330:341-348.

    Not in our library10.1016/j.bbrc.2005.02.175

16. 16.

    Keogh MB, Partap S, Daly JS, et al. Three hours of perfusion culture prior to 28 days of static culture, enhances osteogenesis by human cells in a collagen gag scaffold. Biotechnol Bioeng. 2011;108:1203-1210.

    Not in our library10.1002/bit.23032

17. 17.

    Knippenberg M, Helder MN, Doulabi BZ, et al. Adipose tissue-derived mesenchymal stem cells acquire bone cell-like responsiveness to fluid shear stress on osteogenic stimulation. Tissue engineering. 2005;11:1780-1788.

    Not in our library10.1089/ten.2005.11.1780

18. 18.

    Liu L, Yuan W, Wang J. Mechanisms for osteogenic differentiation of human mesenchymal stem cells induced by fluid shear stress. Biomech Model Mechanobiol. 2010;9:659-670.

    Not in our library10.1007/s10237-010-0206-x

19. 19.

    Scaglione S, Wendt D, Miggino S, et al. Effects of fluid flow and calcium phosphate coating on human bone marrow stromal cells cultured in a defined 2D model system. J Biomed Mater Res. Part A. 2008;86:411-419.

    Not in our library10.1002/jbm.a.31607

20. 20.

    Bjerre L, Bunger C, Baatrup A, et al. Flow perfusion culture of human mesenchymal stem cells on coralline hydroxyapatite scaffolds with various pore sizes. J Biomed Mater Res. Part A. 2011;97:251-263.

    Not in our library10.1002/jbm.a.33051

21. 21.

    Bjerre L, Bunger CE, Kassem M, et al. Flow perfusion culture of human mesenchymal stem cells on silicate-substituted tricalcium phosphate scaffolds. Biomaterials. 2008;29:2616-2627.

    Not in our library10.1016/j.biomaterials.2008.03.003

22. 22.

    Zhao F, Chella R, Ma T. Effects of shear stress on 3-D human mesenchymal stem cell construct development in a perfusion bioreactor system: Experiments and hydrodynamic modeling. Biotechnol Bioeng. 2007;96:584-595.

    Not in our library10.1002/bit.21184

23. 23.

    Tjabringa GS, Vezeridis PS, Zandieh-Doulabi B, et al. Polyamines modulate nitric oxide production and cox-2 gene expression in response to mechanical loading in human adipose tissue-derived mesenchymal stem cells. Stem Cells. 2006;24:2262-2269.

    Not in our library10.1634/stemcells.2005-0625

24. 24.

    Li YJ, Batra NN, You L, et al. Oscillatory fluid flow affects human marrow stromal cell proliferation and differentiation. J Orthop Res. 2004;22:1283-1239.

    Not in our library10.1016/j.orthres.2004.04.002

25. 25.

    Riddle RC, Taylor AF, Rogers JR, et al. Atp release mediates fluid flow-induced proliferation of human bone marrow stromal cells. J Bone Miner Res. 2007;22:589-600.

    Not in our library10.1359/jbmr.070113

26. 26.

    Riddle RC, Taylor AF, Genetos DC, et al. Map kinase and calcium signaling mediate fluid flow-induced human mesenchymal stem cell proliferation. Am J Physiol Cell Physiol. 2006;290:C776-C784.

    Not in our library10.1152/ajpcell.00082.2005

27. 27.

    Singla V, Reiter JF. The primary cilium as the cell's antenna: Signaling at a sensory organelle. Science. 2006;313:629-633.

    Not in our library10.1126/science.1124534

28. 28.

    Praetorius HA, Spring KR. A physiological view of the primary cilium. Annu Rev Physiol. 2005;67:515-529.

    Not in our library10.1146/annurev.physiol.67.040403.101353

29. 29.

    Lee KL, Hoey DA, Jacobs CR. Primary cilia-mediated mechanotransduction in bone. Clin Rev Bone Miner Metab. 2012;8:201-212.

    Not in our library

30. 30.

    Hoey DA, Downs ME, Jacobs CR. The mechanics of the primary cilium: An intricate structure with complex function. J Biomech. 2012;45:17-26.

    Not in our library10.1016/j.jbiomech.2011.08.008

31. 31.

    Ecker A. Flimmerbewegung im Gehörorgan von Petromyzon marinus. Arch Anat Physiol Wiss Med. 1844;520-521.

    Not in our library

32. 32.

    Zimmerman KW. Beitrage zur Kenntniss einiger Drusen und Epithelien. Arch Mikrosk Anat. 1898;52:552-706.

    Not in our library

33. 33.

    Hildebrandt F, Benzing T, Katsanis N. Ciliopathies. N Engl J Med. 2011;364:1533-1543.

    Not in our library10.1056/nejmra1010172

34. 34.

    Berbari NF, O'Connor AK, Haycraft CJ, et al. The primary cilium as a complex signaling center. Curr Biol. 2009;19:R526-R535.

    Not in our library10.1016/j.cub.2009.05.025

35. 35.

    Praetorius HA, Spring KR. Bending the MDCK cell primary cilium increases intracellular calcium. J Membr Biol. 2001;184:71-79.

    Not in our library10.1007/s00232-001-0075-4

36. 36.

    Nauli SM, Alenghat FJ, Luo Y, et al. Polycystins 1 and 2 mediate mechanosensation in the primary cilium of kidney cells. Nat Genet. 2003;33:129-137.

    Not in our library10.1038/ng1076

37. 37.

    Abdul-Majeed S, Moloney BC, Nauli SM. Mechanisms regulating cilia growth and cilia function in endothelial cells. Cell Mol Life Sci. 2012;69:165-173.

    Not in our library10.1007/s00018-011-0744-0

38. 38.

    Masyuk AI, Masyuk TV, Splinter PL, et al. Cholangiocyte cilia detect changes in luminal fluid flow and transmit them into intracellular Ca2+ and CAMP signaling. Gastroenterology. 2006;131:911-920.

    Not in our library10.1053/j.gastro.2006.07.003

39. 39.

    Masyuk AI, Gradilone SA, Banales JM, et al. Cholangiocyte Primary cilia are chemosensory organelles that detect biliary nucleotides via P2Y12 purinergic receptors. Am J Physiol Gastrointest Liver Physiol. 2008;295:G725-G734.

    Not in our library10.1152/ajpgi.90265.2008

40. 40.

    Shao YY, Wang L, Welter JF, et al. Primary cilia modulate Ihh signal transduction in response to hydrostatic loading of growth plate chondrocytes. Bone. 2012;50:79-84.

    Not in our library10.1016/j.bone.2011.08.033

41. 41.

    Malone AM, Anderson CT, Tummala P, et al. Primary cilia mediate mechanosensing in bone cells by a calcium-independent mechanism. Proc Natl Acad Sci USA. 2007;104:13325-13330.

    Not in our library10.1073/pnas.0700636104

42. 42.

    Hoey DA, Kelly DJ, Jacobs CR. A role for the primary cilium in paracrine signaling between mechanically stimulated osteocytes and mesenchymal stem cells. Biochem Biophys Res Commun. 2011;412:182-187.

    Not in our library10.1016/j.bbrc.2011.07.072

43. 43.

    Kwon RY, Temiyasathit S, Tummala P, et al. Primary cilium-dependent mechanosensing is mediated by adenylyl cyclase 6 and cyclic AMP in bone cells. FASEB J. 2010;24:2859-2868.

    Not in our library10.1096/fj.09-148007

44. 44.

    Tummala P, Arnsdorf EJ, Jacobs CR. The role of primary cilia in mesenchymal stem cell differentiation: A pivotal switch in guiding lineage commitment. Cell Mol Bioeng. 2010;3:207-212.

    Not in our library10.1007/s12195-010-0127-x

45. 45.

    Xiao ZS, Quarles LD. Role of the polycytin-primary cilia complex in bone development and mechanosensing. Ann N Y Acad Sci. 2010;1192:410-421.

    Not in our library10.1111/j.1749-6632.2009.05239.x

46. 46.

    Xiao Z, Zhang S, Cao L, et al. Conditional disruption of Pkd1 in osteoblasts results in osteopenia due to direct impairment of bone formation. J Biol Chem. 2010;285:1177-1187.

    Not in our library10.1074/jbc.m109.050906

47. 47.

    Temiyasathit S, Tang WJ, Leucht P, et al. Mechanosensing by the primary cilium: Deletion of Kif3A reduces bone formation due to loading. PLoS One. 2012;7:e33368.

    Not in our library10.1371/journal.pone.0033368

48. 48.

    Bianco P, Riminucci M, Gronthos S, et al. Bone marrow stromal stem cells: Nature, biology, and potential applications. Stem Cells. 2001;19:180-192.

    Not in our library10.1634/stemcells.19-3-180

49. 49.

    Jaiswal N, Haynesworth SE, Caplan AI, et al. Osteogenic differentiation of purified, culture-expanded human mesenchymal stem cells in vitro. J Cell Biochem. 1997;64:295-312.

    Not in our library

50. 50.

    Jacobs CR, Yellowley CE, Davis BR, et al. Differential effect of steady versus oscillating flow on bone cells. J Biomech. 1998;31:969-976.

    Not in our library10.1016/s0021-9290(98)00114-6

51. 51.

    Taulman PD, Haycraft CJ, Balkovetz DF, et al. Polaris, a protein involved in left-right axis patterning, localizes to basal bodies and cilia. Mol Biol Cell. 2001;12:589-599.

    Not in our library10.1091/mbc.12.3.589

52. 52.

    Nowlan NC, Sharpe J, Roddy KA, et al. Mechanobiology of embryonic skeletal development: Insights from animal models. Birth Defects Res C Embryo Today. 2010;90:203-213.

    Not in our library10.1002/bdrc.20184

53. 53.

    Lee HW, Kim SY, Kim AY, et al. Adiponectin stimulates osteoblast differentiation through induction of COX2 in mesenchymal progenitor cells. Stem Cells. 2009;27:2254-2262.

    Not in our library10.1002/stem.144

54. 54.

    Zhang X, Schwarz EM, Young DA, et al. Cyclooxygenase-2 regulates mesenchymal cell differentiation into the osteoblast lineage and is critically involved in bone repair. J Clin Invest. 2002;109:1405-1415.

    Not in our library10.1172/jci15681

55. 55.

    Forwood MR. Inducible cyclo-oxygenase (COX-2) mediates the induction of bone formation by mechanical loading in vivo. J Bone Miner Res. 1996;11:1688-1693.

    Not in our library10.1002/jbmr.5650111112

56. 56.

    Gerasimovskaya EV, Tucker DA, Weiser-Evans M, et al. Extracellular ATP-induced proliferation of adventitial fibroblasts requires phosphoinositide 3-kinase, Akt, mammalian target of rapamycin, and p70 S6 kinase signaling pathways. J Biol Chem. 2005;280:1838-1848.

    Not in our library10.1074/jbc.m409466200

57. 57.

    Lee DY, Li YS, Chang SF, et al. Oscillatory flow-induced proliferation of osteoblast-like cells is mediated by alphavbeta3 and beta1 integrins through synergistic interactions of focal adhesion kinase and Shc with phosphatidylinositol 3-kinase and the Akt/mTOR/p70S6K pathway. J Biol Chem. 2010;285:30-42.

    Not in our library10.1074/jbc.m109.010512

58. 58.

    Cullen DM, Smith RT, Akhter MP. Bone-loading response varies with strain magnitude and cycle number. J Appl Physiol. 2001;91:1971-1976.

    Not in our library10.1152/jappl.2001.91.5.1971

59. 59.

    Plaisant M, Fontaine C, Cousin W, et al. Activation of Hedgehog signaling inhibits osteoblast differentiation of human mesenchymal stem cells. Stem Cells. 2009;27:703-713.

    Not in our library10.1634/stemcells.2008-0888

60. 60.

    Kiprilov EN, Awan A, Desprat R, et al. Human embryonic stem cells in culture possess primary cilia with Hedgehog signaling machinery. J Cell Biol. 2008;180:897-904.

    Not in our library10.1083/jcb.200706028

61. 61.

    Whitfield JF, Chakravarthy BR. The neuronal primary cilium: Driver of neurogenesis and memory formation in the hippocampal dentate gyrus? Cell Signal. 2009;21:1351-1355.

    Not in our library10.1016/j.cellsig.2009.02.013

62. 62.

    Clement CA, Kristensen SG, Mollgard K, et al. The primary cilium coordinates early cardiogenesis and hedgehog signaling in cardiomyocyte differentiation. J Cell Sci. 2009;122(pt 17):3070-3082.

    Not in our library10.1242/jcs.049676

63. 63.

    Nakamura T, Aikawa T, Iwamoto-Enomoto M, et al. Induction of osteogenic differentiation by Hedgehog proteins. Biochem Biophys Res Commun. 1997;237:465-469.

    Not in our library10.1006/bbrc.1997.7156

64. 64.

    James AW, Leucht P, Levi B, et al. Sonic Hedgehog influences the balance of osteogenesis and adipogenesis in mouse adipose-derived stromal cells. Tissue Eng Part A. 2010;16:2605-2616.

    Not in our library10.1089/ten.tea.2010.0048

65. 65.

    Murakami M, Ichisaka T, Maeda M, et al. Mtor is essential for growth and proliferation in early mouse embryos and embryonic stem cells. Mol Cell Biol. 2004;24:6710-67184.

    Not in our library10.1128/mcb.24.15.6710-6718.2004

66. 66.

    Shaw RJ, Cantley LC, Ras. PI(3)K and mTOR signalling controls tumour cell growth. Nature. 2006;441:424-430.

    Not in our library10.1038/nature04869

67. 67.

    Boehlke C, Kotsis F, Patel V, et al. Primary cilia regulate mTORC1 activity and cell size through Lkb1. Nat Cell Biol. 2010;12:1115-1122.

    Not in our library10.1038/ncb2117

68. 68.

    Shillingford JM, Murcia NS, Larson, et al. CH, The Mtor pathway is regulated by polycystin-1, and its inhibition reverses renal cystogenesis in polycystic kidney disease. Proc Natl Acad Sci USA. 2006;103:5466-5471.

    Not in our library10.1073/pnas.0509694103

69. 69.

    Weimbs T. Polycystic kidney disease and renal injury repair: Common pathways, fluid flow, and the function of polycystin-1. Am J Physiol Renal Physiol. 2007;293:F1423-F1432.

    Not in our library10.1152/ajprenal.00275.2007

70. 70.

    Litzenberger JB, Kim JB, Tummala P, et al. Beta1 integrins mediate mechanosensitive signaling pathways in osteocytes. Calcif Tissue Int. 2010;86:325-332.

    Not in our library10.1007/s00223-010-9343-6

71. 71.

    Young SRL, Gerard-O'Riley R, Kim JB, et al. Focal adhesion kinase is important for fluid shear stress-induced mechanotransduction in osteoblasts. J Bone Miner Res. 2009;24:411-424.

    Not in our library10.1359/jbmr.081102

72. 72.

    McGlashan SR, Jensen CG, Poole CA. Localization of extracellular matrix receptors on the chondrocyte primary cilium. J Histochem Cytochem. 2006;54:1005-1014.

    Not in our library10.1369/jhc.5a6866.2006

73. 73.

    Praetorius HA, Frokiaer J, Praetorius J, et al. beta 1-integrin is expressed on the primary cilia of MDCK cells. J Am Soc Nephrology. 2002;13:739a-739a.

    Not in our library

Cited by papers we hold 3

• Mechanical regulation of bone remodeling · 2022
  No notice
• FBW7 couples structural integrity with functional output of primary cilia · 2021
  Not checked
• Molecular mechanosensors in osteocytes · 2020
  No notice

Reads

A person counts once a day per paper. We keep no cookie and no address: a reader is a hash that changes every day. Any month under five reads shows no number.