In vitro models of normal mammary epithelium have correlated increased extracellular matrix (ECM) stiffness with malignant phenotypes. However, the role of increased stiffness in this transformation remains unclear because of difficulties in controlling ECM stiffness, composition and architecture independently. Here we demonstrate that interpenetrating networks of reconstituted basement membrane matrix and alginate can be used to modulate ECM stiffness independently of composition and architecture. We find that, in normal mammary epithelial cells, increasing ECM stiffness alone induces malignant phenotypes but that the effect is completely abrogated when accompanied by an increase in basement-membrane ligands. We also find that the combination of stiffness and composition is sensed through β4 integrin, Rac1, and the PI3K pathway, and suggest a mechanism in which an increase in ECM stiffness, without an increase in basement membrane ligands, prevents normal α6β4 integrin clustering into hemidesmosomes.
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The authors acknowledge the help of A. Li, D. Klumpers, A. Mao and other members of the Mooney lab. The authors also thank J. Brugge (Harvard Medical School) for providing the β4-integrin and Rac1 mutant plasmids, L. Lichten (Qiagen) for help with RNA arrays, Louise Jawerth/Weitz lab for help/use of the rheometer, M. Ericsson and L. Trakimas of the Harvard Medical School EM facility for help with transmission electron microscopy, P. Mali (Harvard Medical School) for discussions, and the Bauer Core for flow sorting. This work was supported by an NIH F32 grant to O.C. (CA153802), fellowships from NSERC and HHMI for S.T.K., fellowships from FCT, FCG and FLAD for C.B.d.C., and NIH (R01EB015498) and MRSEC (DMR-0820484) grants to D.J.M. This work was performed in part at the Center for Nanoscale Systems (CNS), a member of the National Nanotechnology Infrastructure Network (NNIN).
The authors declare no competing financial interests.
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Chaudhuri, O., Koshy, S., Branco da Cunha, C. et al. Extracellular matrix stiffness and composition jointly regulate the induction of malignant phenotypes in mammary epithelium. Nature Mater 13, 970–978 (2014). https://doi.org/10.1038/nmat4009
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