PMID- 28984468 OWN - NLM STAT- MEDLINE DCOM- 20190715 LR - 20240207 IS - 1535-4989 (Electronic) IS - 1044-1549 (Print) IS - 1044-1549 (Linking) VI - 58 IP - 5 DP - 2018 May TI - TGF-beta1 Evokes Human Airway Smooth Muscle Cell Shortening and Hyperresponsiveness via Smad3. PG - 575-584 LID - 10.1165/rcmb.2017-0247OC [doi] AB - Transforming growth factor beta1 (TGF-beta1), a cytokine whose levels are elevated in the airways of patients with asthma, perpetuates airway inflammation and modulates airway structural cell remodeling. However, the role of TGF-beta1 in excessive airway narrowing in asthma, or airway hyperresponsiveness (AHR), remains unclear. In this study, we set out to investigate the direct effects of TGF-beta1 on human airway smooth muscle (HASM) cell shortening and hyperresponsiveness. The dynamics of AHR and single-cell excitation-contraction coupling were measured in human precision-cut lung slices and in isolated HASM cells using supravital microscopy and magnetic twisting cytometry, respectively. In human precision-cut lung slices, overnight treatment with TGF-beta1 significantly augmented basal and carbachol-induced bronchoconstriction. In isolated HASM cells, TGF-beta1 increased basal and methacholine-induced cytoskeletal stiffness in a dose- and time-dependent manner. TGF-beta1-induced single-cell contraction was corroborated by concomitant increases in myosin light chain and myosin phosphatase target subunit 1 phosphorylation levels, which were attenuated by small interfering RNA-mediated knockdown of Smad3 and pharmacological inhibition of Rho kinase. Strikingly, these physiological effects of TGF-beta1 occurred through a RhoA-independent mechanism, with little effect on HASM cell [Ca(2+)](i) levels. Together, our data suggest that TGF-beta1 enhances HASM excitation-contraction coupling pathways to induce HASM cell shortening and hyperresponsiveness. These findings reveal a potential link between airway injury-repair responses and bronchial hyperreactivity in asthma, and define TGF-beta1 signaling as a potential target to reduce AHR in asthma. FAU - Ojiaku, Christie A AU - Ojiaku CA AUID- ORCID: 0000-0002-2592-6710 AD - 1 Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, and. AD - 2 Rutgers Institute for Translational Medicine and Science, Child Health Institute, Rutgers University, New Brunswick, New Jersey; and. FAU - Cao, Gaoyuan AU - Cao G AD - 2 Rutgers Institute for Translational Medicine and Science, Child Health Institute, Rutgers University, New Brunswick, New Jersey; and. FAU - Zhu, Wanqu AU - Zhu W AD - 3 Department of Environmental Health and Engineering, Johns Hopkins Bloomberg School of Public Health, and. FAU - Yoo, Edwin J AU - Yoo EJ AD - 1 Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, and. AD - 2 Rutgers Institute for Translational Medicine and Science, Child Health Institute, Rutgers University, New Brunswick, New Jersey; and. FAU - Shumyatcher, Maya AU - Shumyatcher M AD - 4 Department of Biostatistics, Epidemiology and Informatics, University of Pennsylvania, Philadelphia, Pennsylvania. FAU - Himes, Blanca E AU - Himes BE AD - 4 Department of Biostatistics, Epidemiology and Informatics, University of Pennsylvania, Philadelphia, Pennsylvania. FAU - An, Steven S AU - An SS AD - 3 Department of Environmental Health and Engineering, Johns Hopkins Bloomberg School of Public Health, and. AD - 5 Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland. FAU - Panettieri, Reynold A Jr AU - Panettieri RA Jr AD - 2 Rutgers Institute for Translational Medicine and Science, Child Health Institute, Rutgers University, New Brunswick, New Jersey; and. LA - eng GR - P01 HL114471/HL/NHLBI NIH HHS/United States GR - R01 HL107361/HL/NHLBI NIH HHS/United States GR - R01 HL133433/HL/NHLBI NIH HHS/United States GR - T32 GM008076/GM/NIGMS NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PL - United States TA - Am J Respir Cell Mol Biol JT - American journal of respiratory cell and molecular biology JID - 8917225 RN - 0 (Myosin Light Chains) RN - 0 (SMAD3 protein, human) RN - 0 (Smad3 Protein) RN - 0 (Transforming Growth Factor beta1) RN - 124671-05-2 (RHOA protein, human) RN - EC 2.7.11.1 (rho-Associated Kinases) RN - EC 3.1.3.53 (Myosin-Light-Chain Phosphatase) RN - EC 3.1.3.53 (PPP1R12A protein, human) RN - EC 3.6.5.2 (rhoA GTP-Binding Protein) SB - IM CIN - Am J Respir Cell Mol Biol. 2018 May;58(5):543-544. PMID: 29714630 MH - Asthma/*metabolism/physiopathology MH - Bronchial Hyperreactivity/*metabolism/physiopathology MH - Bronchoconstriction/*drug effects MH - Calcium Signaling/*drug effects MH - Cells, Cultured MH - Excitation Contraction Coupling/drug effects MH - Humans MH - Muscle, Smooth/*drug effects/metabolism/physiopathology MH - Myocytes, Smooth Muscle/*drug effects/metabolism MH - Myosin Light Chains/metabolism MH - Myosin-Light-Chain Phosphatase/metabolism MH - Phosphorylation MH - Smad3 Protein/genetics/*metabolism MH - Transforming Growth Factor beta1/*pharmacology MH - rho-Associated Kinases/metabolism MH - rhoA GTP-Binding Protein/metabolism PMC - PMC5946330 OTO - NOTNLM OT - asthma OT - bronchoconstriction OT - contraction OT - cytokines OT - remodeling EDAT- 2017/10/07 06:00 MHDA- 2019/07/16 06:00 PMCR- 2019/05/01 CRDT- 2017/10/07 06:00 PHST- 2017/10/07 06:00 [pubmed] PHST- 2019/07/16 06:00 [medline] PHST- 2017/10/07 06:00 [entrez] PHST- 2019/05/01 00:00 [pmc-release] AID - 10.1165/rcmb.2017-0247OC [doi] PST - ppublish SO - Am J Respir Cell Mol Biol. 2018 May;58(5):575-584. doi: 10.1165/rcmb.2017-0247OC.