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PPG: Pathophysiology of Alveolar Epithelial Lung Injury

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PI: Jacob I. Sznajder, MD

 

Our Program Project Grant focuses on the alveolar epithelium during lung injury. Acute lung injury (ALI) and the Acute Respiratory Distress Syndrome (ARDS) are defined by impairment in the functions of the alveolar-capillary barrier. Alveolar epithelial cell function and barrier integrity are critical to maintaining normal gas exchange. An increase in the permeability of the alveolar epithelium to solutes and a decrease in the ability to actively clear sodium result in increased edema and impaired gas exchange in the lungs. Some patients with acute lung injury also develop disordered repair of the alveolar epithelium which is accompanied by fibroblast proliferation and collagen deposition. Fibroproliferation is associated with prolonged need for mechanical ventilation and development of the multiple organ dysfunction syndrome, a major cause of death in patients with ALI and ARDS. The alveolar epithelium is therefore a critical target for interventions designed to reduce the nearly 40% mortality observed in the estimated 150,000 patients in the U.S who develop acute lung injury and ARDS each year. The hypotheses which underlie the specific aims and experiments of each individual project and which drive the overall program are as follows:   

  • Resolution of alveolar edema is essential for recovery after acute lung injury. Regional tissue hypoxia in the injured lung promotes endocytosis and ubiquitin mediated degradation of the Na,K-ATPase, impairing alveolar fluid clearance.

  • Lung injury is associated with regional disruption of alveolar epithelial integrity. Adjacent epithelial cells play a role in re-establishing epithelial integrity through a process that requires the dynamic reorganization of the intermediate filament network.

  • Mechanical forces applied to the alveolar epithelium activate signaling pathways through the extracellular matrix including laminin and dystroglycan. Alterations in extracellular matrix stiffness might interfere with these signaling pathways during lung injury and repair.

  • A subset of patients with lung injury go on to develop fibroblast proliferation and collagen deposition. This fibroproliferation is associated with poor outcomes. Preventing the activation of the intrinsic apoptotic pathway in alveolar epithelial cells by inhibiting the Bcl-2 protein Bid can prevent the development of fibrosis. 

The project investigators bring a wealth of experience to the study of the lung epithelium during injury. Over the last funding period, the projects and cores have developed an abundance of new reagents and procedures including intracellular probes, shRNAs, sophisticated gene delivery systems, advanced cell isolation techniques and transgenic/knockout mice. These reagents are not only proving to be invaluable tools for the ongoing aims but also provide the backbone of the new studies proposed in this continuation.

 

Brief history and rationale for the PPG  

Historic models of acute lung injury suggested that lung injury follows an orderly, sequential pattern in which damage to the alveolar capillary membrane accompanied by edema formation was followed by active clearance of edema fluid and subsequent repair of the alveolar-capillary membrane with a varying degree of fibrosis. It is now recognized that these processes occur simultaneously in the lung of patient with ARDS and persist throughout the illness. This shift in thinking about ARDS necessitates a shift in the therapeutic paradigm. To enhance the survival of patients with lung injury, a multi-pronged approach is required that simultaneously improves the clearance of edema fluid (Project 1), promotes alveolar epithelial repair over fibrosis (Project 2) and prevents the ongoing death of alveolar epithelial cells during injury (Project 3).  

In this program project, the investigators have focused on the mechanisms by which the alveolar epithelium contributes to the development of lung injury, edema, and fibrosis. Drs. Sznajder, Dada and Ciechanover (Project 1) will examine the mechanisms that regulate Na+ transport in epithelial cells exposed to hypoxia. Drs. Ridge, Goldman, Gottardi and Ciechanover (Project 2) will examine the role of the intermediate filament network, a critical component of the cell cytoskeleton, in the repair of the wounded epithelium. Drs. Chandel, Budinger, and Kamp (Project 3) will extend their observations that mitochondrial-dependent apoptosis is required for fibrosis after bleomycin induced lung injury with a particular focus on signaling by TGF-β1. These four groups of investigators will be supported by the cores. Dr. Glucksberg will determine the mechanisms by which exposure of epithelial cells to cyclic stretch enhances gene transfer to the alveolar epithelium. The Administrative Core (Core A) will continue to organize the scheduled meetings between the project investigators, schedule and organize outside speakers sponsored by the PPG and facilitate the interaction between project investigators and other core facilities in the University. The Cell Culture Core (Core B) will continue to provide high quality primary alveolar type I and type II cells from rats, mice and humans as well as established alveolar epithelial cell lines to project investigators. The newly established Physiology- Rat/Mouse Core (Core C) will provide blinded measurements of physiologic outcomes in relevant rat and mouse models of lung injury, breed transgenic and wild-type mice for project investigators and create genetic mouse tools for use by the project investigators and by the pulmonary research community. Collectively, these projects will further enhance our understanding of the mechanisms by which the alveolar epithelium contributes to lung injury and provide strategies to promote alveolar epithelial repair in the injured lung.

 

Publications

Wei J, Fang F, Lam AP, Sargent JL, Hamburg E, Hinchcliff ME, Gottardi CJ, Atit R, Whitfield ML, Varga J. 2012. Wnt/β-catenin signaling is hyperactivated in systemic sclerosis and induces Smad-dependent fibrotic responses in mesenchymal cells. Arthritis Rheum. Feb 10. doi: 10.1002/art.34424. [Epub ahead of print] PMID:  22328118. 

Soberanes S, A Gonzalez, D Urich, SE Chiarella, KA Radigan, A Osornio-Vargas, J Joseph, B Kalyanaraman, KM Ridge, NS Chandel, GM Mutlu, A De Vizcaya-Ruiz and GRS Budinger. Particulate matter air pollution induces hypermethylation of the p16 promoter via a mitochondrial ROS-JNK-DNMT1 pathway. Scientific Reports. January 2012.

Mutlu GM*, Budinger GRS*, Wu M, Lam AP, Zirk A, Rivera S, Urich D, Chiarella SE, Go LHT, ghosh AK, Selman M, Pardo A, Varga J, Kamp DW, Chandel NS, Sznajder JI and Jain M.  Proteasomal inhibition after injury prevents fibrosis by modulating TGF-β1 signaling.  Thorax. 2012 Feb;67(2):139-46 *Shared author with equal contribution.

Dos Santos, G., Kutuzov, M., Rogel, M., Baker, M., and Ridge, K.M.  Vimentin is required for LPS-induced inflammasome activation and lung injury.  Am. Thoracic Society, 2012

Dada L., Gonzalez A.R., Urich D., Soberanes S., Manghi T.S., Chiarella S.E., Chandel N.S., Budinger G.R., Mutlu G.M.  Alcohol Worsens Acute Lung Injury by Inhibiting Alveolar Sodium Transport through the Adenosine A1 Receptor. 2012. PLoS One.7:e30448. PMID: 22272351

Zhou Q., Pardo A., Koenigshoff M., Eickelberg O., Budinger G.R.S, Thavarajah K., Gottardi C.J., Jones J., Varga J., Selman M., Sznajder J.I., Raj U., Zhou G. Role of von Hippel-Lindau protein in fibroblast proliferation and fibrosis. 2011. FASEB J, 25:3032-44. PMID: 21642472.

Wei, J., Melichian, D., Komura, K., Hinchcliff, M., Lam, A. P., Lafyatis, R., Gottardi, C. J., MacDougald, O. A. and John Varga. 2011. Canonical Wnt signaling induces skin fibrosis and subcutaneous lipoatrophy: a novel mouse model for scleroderma? Arthritis Rheum. 2011 Jun;63(6):1707-17 PMID: 21370225.

Urich D, Eisenberg JL, Hamill KJ, Takawira D, Chiarella SE, Soberanes S, Gonzalez A, Koentgen F, Manghi T, Hopkinson SB, Misharin AV, Perlman H, Mutlu GM, Budinger GS* and Jones, JCR*.  Lung-specific loss of the lamining α3 subunit confers resistance to mechanical injury.  J Cell Sci. 2011 Sep 1;124(Pt 17):2927-37. *Shared author with equal contribution.

Takawira D, Budinger GR, Hopkinson SB, Jones JC. A dystroglycan/plectin scaffold mediates mechanical pathway bifurcation in lung epithelial cells. J Biol Chem. 2011;286:6301-10.

Rogel, M., Soni, P.N., Troken, J.R., Sitikov, A., Trejo, H.A. and Ridge, K.M. Vimentin is sufficient and required for wound repair and remodeling in alveolar epithelial cells.  FASEB J. 25:3873-83, 2011.

Rogel, M. and Ridge, K.M.  TGF-β1-induced vimentin is sufficient and required for alveolar epithelial wound repair.  Mol. Biol. Cell 22 (suppl), 1616, 2011.

Ridge, K.M.  Intermediate Filaments and Innate Immunity in the Lung.  Mol. Biol. Cell 22 (suppl), 1617, 2011.

Mutlu GM, Bryce PJ, Budinger GR. Linking air pollution exposure with thrombosis. Blood. 2011 Sep 1;118(9):2636-7.

Mutlu EA, Engen PA, Soberanes S, Urich D, Forsyth CB, Nigelioglu R, Chiarella SE, Radigan KA, Gonzalez A, Jakate S, Keshavarzian A, Budinger GRS and Mutlu GM.  Particulate matter air pollution causes oxidant-mediated increase in gut permeability in mice. Part Fibre Toxicol.  2011;8:19.

Lam, A.P., Tuder, R. and C. J. Gottardi. 2011. Regenerative pathways and emphysema: A new paradigm? AJRCCM, Mar 15;183(6):688-90 PMID:  21471062.

Lam, A.P. and C.J. Gottardi. 2011. Beta-catenin signaling: a novel mediator of fibrosis and potential therapeutic target. Current Opinion in Rheumatology. Nov;23(6):562-7. Review.  PMID: 21885974.

Lam AP, Flozak AS, Russell S, Wei J, Jain M, Mutlu GM, Budinger GR, Feghali-Bostwick CA, Varga J, Gottardi CJ. Nuclear β-catenin is Increased in SSc Pulmonary Fibrosis and Promotes Lung Fibroblast Migration and Proliferation. Am J Respir Cell Mol Biol. 2011 Apr 14.

Kidd, M., Rogel, M., and Ridge, K.M.   Invasion of non-small cell lung cancer cells is mediated by vimentin intermediate filaments. In: BMES 2011 Annual Meeting; October 12-15, 2011; Hartford, Connecticut.

Jain, M., Lam, A. and C. J. Gottardi.  2011. Tissue-Specific Knockout/Knockdown of Type 2 TGF-{beta} Receptor and Protection against Bleomycin Injury/Fibrosis. Am J Respir Crit Care Med. 2011 Oct 15;184(8):983. PMID:  22003151.

Jain M*, Budinger GR*, Lo A, Urich D, Rivera SE, Ghosh AK, Gonzalez A, Chiarella SE, Marks K, Donnelly HK, Soberanes S, Varga J, Radigan KA, Chandel NS, Mutlu GM. Leptin promotes fibroproliferative acute respiratory distress syndrome by inhibiting peroxisome proliferator-activated receptor-γ. Am J Respir Crit Care Med. 2011;183:1490-8. *Shared author with equal contribution.

Gusarova G., Dada L.A., Briva A., Trejo H.E, Hamanaka R.B., Mutlu G.M., Chandel N.S., i Prakriya M, Sznajder J.I. Hypoxia leads to Na,K-ATPase down-regulation via Ca2+ release-activated Ca2+ (CRAC) channels and AMPK activation. 2011.Mol Cel Biol. 31:3546-56. PMID: 21730292

Duch MC*, Budinger GR*, Liang YT, Soberanes S, Urich D, Chiarella SE, Campochiaro LA, Gonzalez A, Chandel NS, Hersam MC, Mutlu GM. Minimizing oxidation and stable nanoscale dispersion improves the biocompatibility of graphene in the lung  Nano Lett. 2011 Dec 14;11(12):5201-7. Epub 2011 Oct 27. *Shared author with equal contribution.

Dada, L.A, Sznajder J.I. Mitochondrial Ca2+ and ROS take center stage to orchestrate TNF-α–mediated inflammatory responses 2011. J Clin Invest. 121: 1683–1685. PMCID: PMC3083786

Budinger, GR and Mutlu GM.  Update in Environmental and Occupational Medicine.  Am J Respir Crit Care Med.  2011;183:1614-9.

Budinger GR, Walley KR. Predicting mortality in patients with acute lung injury. Am J Respir Crit Care Med. 2011 Aug 15;184(4):394-5.

Budinger GR, Mutlu GM, Urich D, Soberanes S, Buccellato LJ, Hawkins K, Chiarella SE, Radigan KA, Eisenbart J, Agrawal H, Berkelhamer S, Hekimi S, Zhang J, Perlman H, Schumacker PT, Jain M, Chandel NS. Epithelial Cell Death is an Important Contributor to Oxidant-Mediated Acute Lung Injury. American Journal of Respiratory and Critical Care Medicine. 2011;183:1043-54.

Budinger GR, McKell JL, Urich D, Foiles N, Weiss I, Chiarella SE, Gonzalez A, Soberanes S, Ghio AJ, Nigdelioglu R, Mutlu EA, Radigan KA, Green D, Kwaan HC, Mutlu GM. Particulate matter-induced lung inflammation increases systemic levels of PAI-1 and activates coagulation through distinct mechanisms. PLoS One. 2011;6:e18525

Weinberg F, Hamanaka R, Wheaton WW, Weinberg S, Joseph J, Lopez M, Kalyanaraman B, Mutlu GM, Budinger GR, Chandel NS. Mitochondrial metabolism and ROS generation are essential for Kras-mediated tumorigenicity. Proc Natl Acad Sci U S A. 2010;107:8788-93.

Mutlu GM, Budinger GRS. Not much turbulence: addition of heliox to noninvasive ventilation fails to improve outcomes in patients with exacerbations of chronic obstructive pulmonary disease. Crit Care Med. 2010 38:319-20.

Mutlu GM, Budinger GR, Green AA, Urich D, Soberanes S, Chiarella SE, Alheid GF, McCrimmon DR, Szleifer I, Hersam MC. Biocompatible Nanoscale Dispersion of Single-Walled Carbon Nanotubes Minimizes in vivo Pulmonary Toxicity. Nano Lett. 2010 Apr 8. 2010;10:1664-70.

Flozak, A.S.*, Lam, A.P.*, Russell, S., Jain, M., Peled, O.N., Sheppard, K.A., Beri, R., Mutlu, G.M., Budinger, G.R.S. and C.J. Gottardi.  2010. beta-catenin/TCF signaling is activated during lung injury and promotes the survival and migration of alveolar epithelial cells. J. Biol. Chem. Jan 29; 285(5):3157-67. Epub Nov 20. PMID: 19933277.

Van Hee VC,  Kaufman JD,  Budinger GRS and Mutlu GM. Update in Environmental and Occupational Medicine 2009.  American Journal of Respiratory and Critical Care Medicine. 2010; 181:1174-80.

Gusarova G., L. A Dada, A. Kelly, C. Brodie, L.A. Witters, N.S. Chandel and J.I. Sznajder. α1-AMP-activated kinase (AMPK) regulates hypoxia-induced Na,K-ATPase endocytosis via direct phosphorylation of PKCζ. 2009. Mol. Cell. Biol. 29(13):3455-3464. PMCID: PMC2698765.

Zhou G, L.A. Dada, M. Wu, A. Kelly, H. Trejo, Q. Zhou, J. Varga, J.I. Sznajder. Hypoxia-induced alveolar epithelial-mesenchymal transition requires mitochondrial ROS and hypoxia-inducible factor 1. 2009. Am J Physiol Lung Cell Mol Physiol. 297(6):L1120-30. PMCID: PMC2793183.

Lecuona E, H. Sun, C. Vohwinkel, A. Ciechanover, J.I. Sznajder. Ubiquitination participates in the lysosomal degradation of Na,K-ATPase in steady-state conditions. 2009. Am J Respir Cell Mol Biol. 41(6):671-9. PMCID: PMC2784405

Rogel, M., Glasauer, A., and Ridge, K.M. TGF-β1 increases vimentin expression in alveolar epithelial cells. BMES 2009, Pittsburg, PA.

Flozak, A., Lam, AP, Russell, S., Jain, M., Peled, ON, Sheppard, KA., Beri, R., Mutlu, GM., Budinger, GR, and C.J. Gottardi. Beta-catenin/T-cell factor signaling is activated during lung injury and promotes the survival and migration of alveolar epithelial cells. J. Biol. Chem., 285:3157-3167, 2010. PMCID: PMC2823449.

Rogel, M.R., Jaitovich, A., Ridge, K.M. The role of the ubiquitin proteasome pathway in keratin intermediate filament protein degradation. Proc. Am. Thorac. Soc. 7:71-6, 2010. PMCID: PMC Journal - In Process.

Gökhan M. Mutlu*, G.R. Scott Budinger*,Daniela Urich, Leonard J. Buccellato, Keenan Hawkins, Sergio Chiarella, James Eisenbart, Saul Soberanes, Manu Jain, and Navdeep S. Chandel (2010) Epithelial cell death is an important contributor to oxidant-mediated acute lung injury. Submitted to American Journal of Respiratory Critical Care Medicine.

Weiss CH, Budinger GR, Mutlu GM, Jain M. Proteasomal regulation of pulmonary fibrosis. Proc Am Thorac Soc. 2010 Feb;7(1):77-83. PMCID: PMC Journal - In Process.

Panduri V, Liu G, Surapureddi S, Kondapalli J, Soberanes S, de Souza-Pinto NC, Bohr VA, Budinger GR, Schumacker PT, Weitzman SA, Kamp DW. Role of mitochondrial hOGG1 and aconitase in oxidant-induced lung epithelial cell apoptosis. Free Radic Biol Med. 2009 Sep 15;47(6):750-9. Epub 2009 Jun 12.PMID: 19524665.

Urich D, Soberanes S, Burgess Z, Chiarella SE, Ghio AJ, Ridge KM, Kamp DW, Chandel NS, Mutlu GM, Budinger GR. Proapoptotic Noxa is required for particulate matter-induced cell death and lung inflammation. FASEB J. 2009 Jul;23(7):2055-64. Epub 2009 Feb 23. PMCID: PMC2704586.

Soberanes S, Urich D, Baker CM, Burgess Z, Chiarella SE, Bell EL, Ghio AJ, De Vizcaya-Ruiz A, Liu J, Ridge KM, Kamp DW, Chandel NS, Schumacker PT, Mutlu GM, Budinger GR. Mitochondrial complex III-generated oxidants activate ASK1 and JNK to induce alveolar epithelial cell death following exposure to particulate matter air pollution. J Biol Chem. 2009 Jan 23;284(4):2176-86. Epub 2008 Nov 25. PMCID: PMC2629089.
 

Abstracts

Rogel, M., and Ridge, K.M.   Vimentin is sufficient and required for wound repair and remodeling in alveolar epithelial cells. In: BMES 2011 Annual Meeting; October 12-15, 2011; Hartford, Connecticut.

L.A. Dada, PhD, G. Gusarova, PhD, A. Kelly, BS, F. Moazed, MD, M. Baker, J.I. Sznajder, MD, Chicago, IL. AMP-Activated Protein Kinase (AMPK) Regulates the Hypoxia-Induced Na,K-ATPase Endocytosis Via Direct Phosphorylation of PKCζ, [Publication Page: A1926]

G.A. Gusarova, PhD, L.A. Dada, PhD, A. Briva, MD, H. Trejo, MD, J.I. Sznajder, MD, Chicago, IL. Hypoxia Leads to Ca2+/Calmodulin-Dependent Kinase Kinase β (CaMKKβ) Activation/AMPK Phosphorylation, Na,K-ATPase Endocytosis and Impaired Alveolar Epithelial Reabsorbtion, [Publication Page: A1927]