Abstract
The regenerative capacity of the mammary gland following post-lactational involution depends on the presence of multipotent stem or progenitor cells. Mammary progenitor cells exist as a quiescent and self-renewing population capable of differentiating into luminal epithelial and myoepithelial cells and generating ductal and alveolar structures. The fate choices of these cells are regulated by several soluble signals as well as their surrounding extracellular matrix. Whereas matrix stiffness has been implicated in organ-specific differentiation of embryonic and mesenchymal stem cells, the effects of substratum compliance on the more limited fate switches typical of tissue-specific progenitor cells are unknown. Here, we examined how the mechanical properties of the microenvironment affect the differentiation of mammary progenitor cells. Immortalized human mammary progenitor cells were cultured on synthetic hydrogels of varying stiffness, and their self-renewal and fate decisions were quantified. We found that cells cultured on soft substrata differentiated preferentially into luminal epithelial cells, whereas those cultured on stiff substrata differentiated preferentially into myoepithelial cells. Furthermore, pharmacological manipulations of cytoskeletal tension in conjunction with analysis of gene expression revealed that mechanical properties of the microenvironment signal through the small GTPase RhoA and cytoskeletal contractility to modulate the differentiation of mammary progenitor cells. These data suggest that subtle variations in the mechanical compliance of a tissue can direct the fate decisions of its resident progenitor cells.
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Abbreviations
- ECM:
-
Extracellular matrix
- ES:
-
Embryonic stem
- FAK:
-
Focal adhesion kinase
- MLCK:
-
Myosin light chain kinase
- MSCs:
-
Mesenchymal stem cells
- PA:
-
Polyacrylamide
- ROCK:
-
Rho-associated kinase
- TDLU:
-
Terminal ductal lobular unit
References
Alcaraz J, Xu R, Mori H, Nelson CM, Mroue R, Spencer VA, Brownfield D, Radisky DC, Bustamante C, Bissell MJ (2008) Laminin and biomimetic extracellular elasticity enhance functional differentiation in mammary epithelia. EMBO J 27(21): 2829–2838
Alenghat FJ, Ingber DE (2002) Mechanotransduction: all signals point to cytoskeleton, matrix, and integrins. Sci STKE [electronic resource]: signal transduction knowledge environment (119)
Amano M, Ito M, Kimura K, Fukata Y, Chihara K, Nakano T, Matsuura Y, Kaibuchi K (1996) Phosphorylation and activation of myosin by rho-associated kinase (rho-kinase). J Biol Chem 271(34): 20246–20249. doi:10.1074/jbc.271.34.20246
Anderson LH, Boulanger CA, Smith GH, Carmeliet P, Watson CJ (2011) Stem cell marker prominin-1 regulates branching morphogenesis, but not regenerative capacity, in the mammary gland. Dev Dyn 240(3): 674–681. doi:10.1002/dvdy.22539
Batistatou A, Stefanou D, Arkoumani E, Agnantis NJ (2003) The usefulness of p63 as a marker of breast myoepithelial cells. In Vivo 17(6): 573–576
Beningo KA, Dembo M, Kaverina I, Small JV, Wang Y-l (2001) Nascent focal adhesions are responsible for the generation of strong propulsive forces in migrating fibroblasts. J Cell Biol 153(4): 881–888. doi:10.1083/jcb.153.4.881
Bershadsky AD, Balaban NQ, Geiger B (2003) Adhesion-dependent cell mechanosensitivity. Annu Rev Cell Dev Biol 19(1): 677–695. doi:10.1146/annurev.cellbio.19.111301.153011
Boudou T, Ohayon J, Picart C, Tracqui P (2006) An extended relationship for the characterization of Young’s modulus and Poisson’s ratio of tunable polyacrylamide gels. Biorheology 43(6): 721–728
Davies SP, Reddy H, Caivano M, Cohen P (2000) Specificity and mechanism of action of some commonly used protein kinase inhibitors. Biochem J 351(Pt 1): 95–105
Dontu G, Jackson KW, McNicholas E, Kawamura MJ, Abdallah WM, Wicha MS (2004) Role of Notch signaling in cell-fate determination of human mammary stem/progenitor cells. Breast Cancer Res 6(6): R605–615
Emerman JT, Burwen SJ, Pitelka DR (1979) Substrate properties influencing ultrastructural differentiation of mammary epithelial cells in culture. Tissue Cell 11(1): 109–119
Engler AJ, Sen S, Sweeney HL, Discher DE (2006) Matrix elasticity directs stem cell lineage specification. Cell 126(4): 677–689. doi:10.1016/j.cell.2006.06.044
Evans ND, Minelli C, Gentleman E, LaPointe V, Patankar SN, Kallivretaki M, Chen X, Roberts CJ, Stevens MM (2009) Substrate stiffness affects early differentiation events in embryonic stem cells. Eur Cell Mater 18: 1–13 (discussion 13–14)
Eyckmans J, Boudou T, Yu X, Chen CS (2011) A hitchhiker’s guide to mechanobiology. Dev Cell 21(1): 35–47
Gjorevski N, Nelson CM (2010) Endogenous patterns of mechanical stress are required for branching morphogenesis. Integr Biol Camb 2(9): 424–434
Gjorevski N, Nelson CM (2011) Integrated morphodynamic signalling of the mammary gland. Nat Rev Mol Cell Biol 12: 581–593
Ingber DE (2004) The mechanochemical basis of cell and tissue regulation. Mech Chem Biosyst MCB 1(1): 53–68
Ishihara H, Martin BL, Brautigan DL, Karaki H, Ozaki H, Kato Y, Fusetani N, Watabe S, Hashimoto K, Uemura D et al (1989) Calyculin A and okadaic acid: inhibitors of protein phosphatase activity. Biochem Biophys Res Commun 159(3): 871–877
Ishizaki T, Naito M, Fujisawa K, Maekawa M, Watanabe N, Saito Y, Narumiya S (1997) p160ROCK, a Rho-associated coiled-coil forming protein kinase, works downstream of Rho and induces focal adhesions. FEBS Lett 404(2–3): 118–124. doi:10.1016/s0014-5793(97)00107-5
Kimura K, Ito M, Amano M, Chihara K, Fukata Y, Nakafuku M, Yamamori B, Feng J, Nakano T, Okawa K, Iwamatsu A, Kaibuchi K (1996) Regulation of myosin phosphatase by rho and rho-associated kinase (rho-kinase). Science 273(5272): 245–248. doi:10.1126/science.273.5272.245
Korkaya H, Paulson A, Charafe-Jauffret E, Ginestier C, Brown M, Dutcher J, Clouthier SG, Wicha MS (2009) Regulation of mammary stem/progenitor cells by PTEN/Akt/beta-catenin signaling. PLoS Biol 7(6): e1000121
LaBarge MA, Nelson CM, Villadsen R, Fridriksdottir A, Ruth JR, Stampfer MR, Petersen OW, Bissell MJ (2009) Human mammary progenitor cell fate decisions are products of interactions with combinatorial microenvironments. Integr Biol 1: 70–79
Levental KR, Yu H, Kass L, Lakins JN, Egeblad M, Erler JT, Fong SF, Csiszar K, Giaccia A, Weninger W, Yamauchi M, Gasser DL, Weaver VM (2009) Matrix crosslinking forces tumor progression by enhancing integrin signaling. Cell 139: 891–906
Lim E, Wu D, Pal B, Bouras T, Asselin-Labat ML, Vaillant F, Yagita H, Lindeman GJ, Smyth GK, Visvader JE (2010) Transcriptome analyses of mouse and human mammary cell subpopulations reveal multiple conserved genes and pathways. Breast Cancer Res 12(2): R21
Liu S, Dontu G, Mantle ID, Patel S, Ahn NS, Jackson KW, Suri P, Wicha MS (2006) Hedgehog signaling and Bmi-1 regulate self-renewal of normal and malignant human mammary stem cells. Cancer Res 66(12): 6063–6071
Lorenzen J, Sinkus R, Biesterfeldt M, Adam G (2003) Menstrual-cycle dependence of breast parenchyma elasticity: estimation with magnetic resonance elastography of breast tissue during the menstrual cycle. Invest Radiol 38(4): 236–240. doi:10.1097/01.RLI.0000059544.18910.BD
Lorenzen J, Sinkus R, Lorenzen M, Dargatz M, Leussler C, Roschmann P, Adam G (2002) MR elastography of the breast:preliminary clinical results. Rofo 174(7): 830–834. doi:10.1055/s-2002-32690
McBeath R, Pirone DM, Nelson CM, Bhadriraju K, Chen CS (2004) Cell shape, cytoskeletal tension, and RhoA regulate stem cell lineage commitment. Dev Cell 6(4): 483–495. doi:10.1016/s1534-5807(04)00075-9
Mitra SK, Hanson DA, Schlaepfer DD (2005) Focal adhesion kinase: in command and control of cell motility. Nat Rev Mol Cell Biol 6(1): 56–68
Paszek MJ, Zahir N, Johnson KR, Lakins JN, Rozenberg GI, Gefen A, Reinhart-King CA, Margulies SS, Dembo M, Boettiger D, Hammer DA, Weaver VM (2005) Tensional homeostasis and the malignant phenotype. Cancer Cell 8(3): 241–254
Pelham RJ, Wang Y-l (1997) Cell locomotion and focal adhesions are regulated by substrate flexibility. Proc Natl Acad Sci USA 94(25): 13661–13665
Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284(5411): 143–147
Samani A, Zubovits J, Plewes D (2007) Elastic moduli of normal and pathological human breast tissues: an inversion-technique-based investigation of 169 samples. Phys Med Biol 52(6): 1565–1576
Schwartz MA (2010) Integrins and extracellular matrix in mechanotransduction. Cold Spring Harb Perspect Biol 2(12): a005066
Srivastava A, Verma Y, Rao KD, Gupta PK (2011) Determination of elastic properties of resected human breast tissue samples using optical coherence tomographic elastography. Strain 47: 75–87
Sternlicht MD, Lochter A, Sympson CJ, Huey B, Rougier JP, Gray JW, Pinkel D, Bissell MJ, Werb Z (1999) The stromal proteinase MMP3/stromelysin-1 promotes mammary carcinogenesis. Cell 98(2): 137–146
Straight AF, Cheung A, Limouze J, Chen I, Westwood NJ, Sellers JR, Mitchison TJ (2003) Dissecting temporal and spatial control of cytokinesis with a myosin II inhibitor. Science 299(5613): 1743–1747
Taddei I, Deugnier MA, Faraldo MM, Petit V, Bouvard D, Medina D, Fassler R, Thiery JP, Glukhova MA (2008) Beta1 integrin deletion from the basal compartment of the mammary epithelium affects stem cells. Nat Cell Biol 10(6): 716–722
Tamada M, Sheetz MP, Sawada Y (2004) Activation of a signaling ascade by cytoskeleton stretch. Dev Cell 7(5): 709–718. doi:10.1016/j.devcel.2004.08.021
Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshall VS, Jones JM (1998) Embryonic stem cell lines derived from human blastocysts. Science 282(5391): 1145–1147
Tiede B, Kang Y (2011) From milk to malignancy: the role of mammary stem cells in development, pregnancy and breast cancer. Cell Res 21(2): 245–257
Villadsen R, Fridriksdottir AJ, Ronnov-Jessen L, Gudjonsson T, Rank F, LaBarge MA, Bissell MJ, Petersen OW (2007) Evidence for a stem cell hierarchy in the adult human breast. J Cell Biol 177(1): 87–101
Visvader JE, Smith GH (2011) Murine mammary epithelial stem cells: discovery, function, and current status. Cold Spring Harb Perspect Biol 3(2): a004879
Woodward WA, Chen MS, Behbod F, Rosen JM (2005) On mammary stem cells. J Cell Sci 118(Pt 16): 3585–3594. doi:10.1242/jcs.02532
Wozniak MA, Desai R, Solski PA, Der CJ, Keely PJ (2003) ROCK-generated contractility regulates breast epithelial cell differentiation in response to the physical properties of a three-dimensional collagen matrix. J Cell Biol 163(3): 583–595
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Lui, C., Lee, K. & Nelson, C.M. Matrix compliance and RhoA direct the differentiation of mammary progenitor cells. Biomech Model Mechanobiol 11, 1241–1249 (2012). https://doi.org/10.1007/s10237-011-0362-7
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DOI: https://doi.org/10.1007/s10237-011-0362-7