Daniel R Potter1, Byron Y Miyazawa, Stuart L Gibb, Xutao Deng, Padma P Togaratti, Roxanne H Croze, Amit K Srivastava, Alpa Trivedi, Michael Matthay, John B Holcomb, Martin A Schreiber, Shibani Pati. 1. From The Department of Laboratory Medicine (D.R.P., B.Y.M., S.L.G., A.T., S.P.), University of California at San Francisco; Blood Systems Research Institute (X.D., P.P.T., S.P.), San Francisco, California; Department of Pediatric Surgery (A.K.S.), McGovern Medical School, University of Texas Health Sciences Center at Houston, Houston, Texas; Cardiovascular Research Institute (R.H.C.), University of California at San Francisco, San Francisco, California; Department of Surgery (J.B.H.), University of Texas, Houston, Texas; and Department of Surgery (M.A.S.), Oregon Health & Science University, Portland, Oregon.
Abstract
BACKGROUND: Mesenchymal stem cells (MSCs) have been shown to mitigate vascular permeability in hemorrhagic shock (HS) and trauma-induced brain and lung injury. Mechanistically, paracrine factors secreted from MSCs have been identified that can recapitulate many of the potent biologic effects of MSCs in animal models of disease. Interestingly, MSC-derived extracellular vesicles (EVs), contain many of these key soluble factors, and have therapeutic potential independent of the parent cells. In this study we sought to determine whether MSC-derived EVs (MSC EVs) could recapitulate the beneficial therapeutic effects of MSCs on lung vascular permeability induced by HS in mice. METHODS: Mesenchymal stem cell EVs were isolated from human bone marrow-derived MSCs by ultracentrifugation. A mouse model of fixed pressure HS was used to study the effects of shock, shock + MSCs and shock + MSC EVs on lung vascular endothelial permeability. Mice were administered MSCs, MSC EVs, or saline IV. Lung tissue was harvested and assayed for permeability, RhoA/Rac1 activation, and for differential phosphoprotein expression. In vitro, human lung microvascular cells junctional integrity was evaluated by immunocytochemistry and endothelial cell impedance assays. RESULTS: Hemorrhagic shock-induced lung vascular permeability was significantly decreased by both MSC and MSC EV infusion. Phosphoprotein profiling of lung tissue revealed differential activation of proteins and pathways related to cytoskeletal rearrangement and regulation of vascular permeability by MSCs and MSC EVs. Lung tissue from treatment groups demonstrated decreased activation of the cytoskeletal GTPase RhoA. In vitro, human lung microvascular cells, MSC CM but not MSC-EVs prevented thrombin-induced endothelial cell permeability as measured by electrical cell-substrate impedance sensing system and immunocytochemistry of VE-cadherin and actin. CONCLUSION: Mesenchymal stem cells and MSC EVs modulate cytoskeletal signaling and attenuate lung vascular permeability after HS. Mesenchymal stem cell EVs may potentially be used as a novel "stem cell free" therapeutic to treat HS-induced lung injury.
BACKGROUND: Mesenchymal stem cells (MSCs) have been shown to mitigate vascular permeability in hemorrhagic shock (HS) and trauma-induced brain and lung injury. Mechanistically, paracrine factors secreted from MSCs have been identified that can recapitulate many of the potent biologic effects of MSCs in animal models of disease. Interestingly, MSC-derived extracellular vesicles (EVs), contain many of these key soluble factors, and have therapeutic potential independent of the parent cells. In this study we sought to determine whether MSC-derived EVs (MSC EVs) could recapitulate the beneficial therapeutic effects of MSCs on lung vascular permeability induced by HS in mice. METHODS: Mesenchymal stem cell EVs were isolated from human bone marrow-derived MSCs by ultracentrifugation. A mouse model of fixed pressure HS was used to study the effects of shock, shock + MSCs and shock + MSC EVs on lung vascular endothelial permeability. Mice were administered MSCs, MSC EVs, or saline IV. Lung tissue was harvested and assayed for permeability, RhoA/Rac1 activation, and for differential phosphoprotein expression. In vitro, human lung microvascular cells junctional integrity was evaluated by immunocytochemistry and endothelial cell impedance assays. RESULTS:Hemorrhagic shock-induced lung vascular permeability was significantly decreased by both MSC and MSC EV infusion. Phosphoprotein profiling of lung tissue revealed differential activation of proteins and pathways related to cytoskeletal rearrangement and regulation of vascular permeability by MSCs and MSC EVs. Lung tissue from treatment groups demonstrated decreased activation of the cytoskeletal GTPase RhoA. In vitro, human lung microvascular cells, MSC CM but not MSC-EVs prevented thrombin-induced endothelial cell permeability as measured by electrical cell-substrate impedance sensing system and immunocytochemistry of VE-cadherin and actin. CONCLUSION: Mesenchymal stem cells and MSC EVs modulate cytoskeletal signaling and attenuate lung vascular permeability after HS. Mesenchymal stem cell EVs may potentially be used as a novel "stem cell free" therapeutic to treat HS-induced lung injury.
Authors: Daniel R Potter; Gail Baimukanova; Sheila M Keating; Xutao Deng; Jeffrey A Chu; Stuart L Gibb; Zhanglong Peng; Marcus O Muench; Marina E Fomin; Philip C Spinella; Rosemary Kozar; Shibani Pati Journal: J Trauma Acute Care Surg Date: 2015-06 Impact factor: 3.313
Authors: Barbara A Christy; Maryanne C Herzig; Robbie K Montgomery; Christopher Delavan; James A Bynum; Kristin M Reddoch; Andrew P Cap Journal: J Trauma Acute Care Surg Date: 2017-07 Impact factor: 3.313
Authors: Jennifer G Wilson; Kathleen D Liu; Hanjing Zhuo; Lizette Caballero; Melanie McMillan; Xiaohui Fang; Katherine Cosgrove; Rosemary Vojnik; Carolyn S Calfee; Jae-Woo Lee; Angela J Rogers; Joseph Levitt; Jeanine Wiener-Kronish; Ednan K Bajwa; Andrew Leavitt; David McKenna; B Taylor Thompson; Michael A Matthay Journal: Lancet Respir Med Date: 2014-12-17 Impact factor: 30.700
Authors: Aarif Y Khakoo; Shibani Pati; Stasia A Anderson; William Reid; Mohamed F Elshal; Ilsa I Rovira; Ahn T Nguyen; Daniela Malide; Christian A Combs; Gentzon Hall; Jianhu Zhang; Mark Raffeld; Terry B Rogers; William Stetler-Stevenson; Joseph A Frank; Marvin Reitz; Toren Finkel Journal: J Exp Med Date: 2006-04-24 Impact factor: 14.307
Authors: Shibani Pati; Marcello Pilia; Juanita M Grimsley; Alexia T Karanikas; Blessing Oyeniyi; John B Holcomb; Andrew P Cap; Todd E Rasmussen Journal: Shock Date: 2015-12 Impact factor: 3.454