title: Integrins beta1, alpha6, and alpha3 contribute to mechanical strain-induced differentiation of fetal lung type II epithelial cells via distinct mechanisms.
abstract: Mechanical forces regulate lung maturation in the fetus by promoting type II epithelial differentiation. However, the cell surface receptors that transduce these mechanical cues into cellular responses remain largely unknown. When distal lung type II epithelial cells isolated from embryonic day 19 rat fetuses were cultured on flexible plates coated with laminin, fibronectin, vitronectin, collagen, or elastin and exposed to a level of mechanical strain (5%) similar to that observed in utero, transmembrane signaling responses were induced under all conditions, as measured by ERK activation. However, mechanical stress maximally increased expression of the type II cell differentiation marker surfactant protein C when cells were cultured on laminin substrates. Strain-induced alveolar epithelial differentiation was inhibited by interfering with cell binding to laminin using soluble laminin peptides (IKVIV or YIGSR) or blocking antibodies against integrin beta1, alpha3, or alpha6. Additional studies were carried out with substrates coated directly with different nonactivating anti-integrin antibodies. Blocking integrin beta1 and alpha6 binding sites inhibited both cell adhesion and differentiation, whereas inhibition of alpha3 prevented differentiation without altering cell attachment. These data demonstrate that various integrins contribute to mechanical control of type II lung epithelial cell differentiation on laminin substrates. However, they may act via distinct mechanisms, including some that are independent of their cell anchoring role.
authors.affiliation: Dept. of Pediatrics, Women and Infants Hospital of Rhode Island, 101 Dudley St., and Department of Medicine, Brown Medical School, Providence, RI 02905, USA. jsanchezesteban@wihri.org
authors.forename: Juan
authors.initials: J
authors.lastname: Sanchez-Esteban
authors.forename: Yulian
authors.initials: Y
authors.lastname: Wang
authors.forename: Edward J
authors.initials: EJ
authors.lastname: Filardo
authors.forename: Lewis P
authors.initials: LP
authors.lastname: Rubin
authors.forename: Donald E
authors.initials: DE
authors.lastname: Ingber
mesh_terms: D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D002454:Cell Differentiation / Q000502:physiology*; D004789:Enzyme Activation; D004847:Epithelial Cells / Q000166:cytology*; D005109:Extracellular Matrix / Q000502:physiology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005333:Fetus / Q000166:cytology; D039422:Integrin alpha3 / Q000502:physiology*; D039503:Integrin alpha6 / Q000502:physiology*; D019012:Integrin beta1 / Q000502:physiology*; D007797:Laminin / Q000502:physiology; D008168:Lung / Q000166:cytology*; D010455:Peptides / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013314:Stress, Mechanical
chemical_list: D039422:Integrin alpha3; D039503:Integrin alpha6; D019012:Integrin beta1; D007797:Laminin; D010455:Peptides; C077063:Sftpc protein, rat; D048049:Extracellular Signal-Regulated MAP Kinases