Confinement Sensing and Signal Optimization via Piezo1/PKA and Myosin II Pathways
Wei-Chien Hung,
Jessica R. Yang,
Christopher L. Yankaskas,
Bin Sheng Wong,
Pei-Hsun Wu,
Carlos Pardo-Pastor,
Selma A. Serra,
Meng-Jung Chiang,
Zhizhan Gu,
Denis Wirtz,
Miguel A. Valverde,
Joy T. Yang,
Jin Zhang,
Konstantinos Konstantopoulos
Affiliations
Wei-Chien Hung
Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD 21218, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA
Jessica R. Yang
Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA
Christopher L. Yankaskas
Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD 21218, USA
Bin Sheng Wong
Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD 21218, USA
Pei-Hsun Wu
Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD 21218, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA
Carlos Pardo-Pastor
Laboratory of Molecular Physiology and Channelopathies, Department of Experimental and Health Sciences, Universitat Pompeu Fabra, Carrera del Doctor Aiguader 88, Barcelona 08003, Spain
Selma A. Serra
Laboratory of Molecular Physiology and Channelopathies, Department of Experimental and Health Sciences, Universitat Pompeu Fabra, Carrera del Doctor Aiguader 88, Barcelona 08003, Spain
Meng-Jung Chiang
Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA
Zhizhan Gu
Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD 21218, USA
Denis Wirtz
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
Miguel A. Valverde
Laboratory of Molecular Physiology and Channelopathies, Department of Experimental and Health Sciences, Universitat Pompeu Fabra, Carrera del Doctor Aiguader 88, Barcelona 08003, Spain
Joy T. Yang
Department of Cell Biology, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Corresponding author
Jin Zhang
Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Corresponding author
Konstantinos Konstantopoulos
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; Corresponding author
Summary: 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 Ca2+ level is increased, leading to phosphodiesterase 1 (PDE1)-dependent suppression of PKA activity. This Ca2+ 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. : Hung et al. demonstrate that a Piezo1-dependent intracellular calcium increase negatively regulates protein kinase A (PKA) as cells transit from unconfined to confined spaces. The Piezo1/PKA and myosin II signaling modules constitute two confinement-sensing mechanisms. This study provides a paradigm by which signaling enables cells to sense and adapt to different microenvironments.
