Inhibition of nuclear factor of activated T cells (NFAT) c3 activation attenuates acute lung injury and pulmonary edema in murine models of sepsis

// Manjula Karpurapu 1 , Yong Gyu Lee 1 , Ziqing Qian 2 , Jin Wen 2 , Megan N. Ballinger 1 , Luiza Rusu 1 , Sangwoon Chung 1 , Jing Deng 1 , Feng Qian 1 , Brenda F. Reader 1 , Teja Srinivas Nirujogi 1 , Gye Young Park 3 , Dehua Pei 2 and John W. Christman 1 1 Pulmonary, Allergy, Critical Care and Sleep Medicine, Ohio State University Wexner Medical Center, Davis Heart and Lung Research Institute, Columbus, OH 43210, USA 2 Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA 3 Pulmonary, Allergy, Critical Care and Sleep Medicine, University of Illinois, Chicago, IL 60612, USA Correspondence to: John W. Christman, email: John.Christman@osumc.edu Manjula Karpurapu, email: Manjula.Karpurapu@osumc.edu Keywords: macrophage; NFAT’s; acute lung injury; pulmonary edema; CP9-ZIZIT Received: September 27, 2016      Accepted: December 22, 2017      Published: January 25, 2018 ABSTRACT Specific therapies targeting cellular and molecular events of sepsis induced Acute Lung Injury (ALI) pathogenesis are lacking. We have reported a pivotal role for Nuclear Factors of Activated T cells (NFATc3) in regulating macrophage phenotype during sepsis induced ALI and subsequent studies demonstrate that NFATc3 transcriptionally regulates macrophage CCR2 and TNFα gene expression. Mouse pulmonary microvascular endothelial cell monolayer maintained a tighter barrier function when co-cultured with LPS stimulated NFATc3 deficient macrophages whereas wild type macrophages caused leaky monolayer barrier. More importantly, NFATc3 deficient mice showed decreased neutrophilic lung inflammation, improved alveolar capillary barrier function, arterial oxygen saturation and survival benefit in lethal CLP sepsis mouse models. In addition, survival of wild type mice subjected to the lethal CLP sepsis was not improved with broad-spectrum antibiotics, whereas the survival of NFATc3 deficient mice was improved to 40–60% when treated with imipenem. Passive adoptive transfer of NFATc3 deficient macrophages conferred protection against LPS induced ALI in wild type mice. Furthermore, CP9-ZIZIT, a highly potent, cell-permeable peptide inhibitor of Calcineurin inhibited NFATc3 activation. CP9-ZIZIT effectively reduced sepsis induced inflammatory cytokines and pulmonary edema in mice. Thus, this study demonstrates that inhibition of NFATc3 activation by CP9-ZIZIT provides a potential therapeutic option for attenuating sepsis induced ALI/pulmonary edema.