title: Confinement Sensing and Signal Optimization via Piezo1/PKA and Myosin II Pathways.
abstract: Cells adopt distinct signaling pathways to optimize cell locomotion in different physical microenvironments. However, the underlying mechanism that enables cells to sense and respond to physical confinement is unknown. Using microfabricated devices and substrate-printing methods along with FRET-based biosensors, we report that, as cells transition from unconfined to confined spaces, intracellular Ca(2+) level is increased, leading to phosphodiesterase 1 (PDE1)-dependent suppression of PKA activity. This Ca(2+) elevation requires Piezo1, a stretch-activated cation channel. Moreover, differential regulation of PKA and cell stiffness in unconfined versus confined cells is abrogated by dual, but not individual, inhibition of Piezo1 and myosin II, indicating that these proteins can independently mediate confinement sensing. Signals activated by Piezo1 and myosin II in response to confinement both feed into a signaling circuit that optimizes cell motility. This study provides a mechanism by which confinement-induced signaling enables cells to sense and adapt to different physical microenvironments.
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD 21218, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA.
authors.forename: Wei-Chien
authors.initials: WC
authors.lastname: Hung
authors.affiliation: Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
authors.forename: Jessica R
authors.initials: JR
authors.lastname: Yang
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD 21218, USA.
authors.forename: Christopher L
authors.initials: CL
authors.lastname: Yankaskas
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD 21218, USA.
authors.forename: Bin Sheng
authors.initials: BS
authors.lastname: Wong
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD 21218, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA.
authors.forename: Pei-Hsun
authors.initials: PH
authors.lastname: Wu
authors.affiliation: Laboratory of Molecular Physiology and Channelopathies, Department of Experimental and Health Sciences, Universitat Pompeu Fabra, Carrera del Doctor Aiguader 88, Barcelona 08003, Spain.
authors.forename: Carlos
authors.initials: C
authors.lastname: Pardo-Pastor
authors.affiliation: Laboratory of Molecular Physiology and Channelopathies, Department of Experimental and Health Sciences, Universitat Pompeu Fabra, Carrera del Doctor Aiguader 88, Barcelona 08003, Spain.
authors.forename: Selma A
authors.initials: SA
authors.lastname: Serra
authors.affiliation: Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
authors.forename: Meng-Jung
authors.initials: MJ
authors.lastname: Chiang
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD 21218, USA.
authors.forename: Zhizhan
authors.initials: Z
authors.lastname: Gu
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD 21218, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA; Department of Oncology, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
authors.forename: Denis
authors.initials: D
authors.lastname: Wirtz
authors.affiliation: Laboratory of Molecular Physiology and Channelopathies, Department of Experimental and Health Sciences, Universitat Pompeu Fabra, Carrera del Doctor Aiguader 88, Barcelona 08003, Spain.
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Valverde
authors.affiliation: Department of Cell Biology, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. Electronic address: jyang@jhmi.edu.
authors.forename: Joy T
authors.initials: JT
authors.lastname: Yang
authors.affiliation: Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. Electronic address: jzhang32@ucsd.edu.
authors.forename: Jin
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD 21218, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA; Department of Oncology, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. Electronic address: konstant@jhu.edu.
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Konstantopoulos
mesh_terms: D000818:Animals; D016466:CHO Cells; D002118:Calcium / Q000378:metabolism / Q000494:pharmacology; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D002465:Cell Movement; D006224:Cricetinae; D003412:Cricetulus; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D054677:Cyclic Nucleotide Phosphodiesterases, Type 1 / Q000378:metabolism; D042541:Intracellular Space / Q000378:metabolism; D007473:Ion Channels / Q000378:metabolism*; D040542:Mechanotransduction, Cellular / Q000187:drug effects; D008545:Melanoma / Q000378:metabolism / Q000473:pathology; D024462:Myosin Type II / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: D007473:Ion Channels; C515204:PIEZO1 protein, human; D017868:Cyclic AMP-Dependent Protein Kinases; D054677:Cyclic Nucleotide Phosphodiesterases, Type 1; D024462:Myosin Type II; D002118:Calcium