Hematopoietic stem cell transplantation (HSCT) is used to treat a variety of genetic defects and malignancies, but its usefulness is limited by pulmonary infections. Infectious complications can occur both in allogeneic and autologous transplant settings and susceptibility to infection remains elevated despite hematopoietic reconstitution. To better understand innate immune deficiencies that characterize HSCT, we developed a murine model of bacterial infection post-syngeneic (syn) bone marrow transplant (BMT). We have previously shown that these mice are more susceptible to infection with Pseudomonas aeruginosa even after the hematopoietic system is reconstituted. We identified the upregulation of cyclooxygenase-2 (COX-2) and the overproduction of PGE2 as major contributing factors to the impaired innate immune function in these mice. We identified that alveolar macrophages (AMs) and neutrophils (PMNs) had defects in innate immune functions such as phagocytosis, bacterial killing and cytokine secretion. In addition, the profile of scavenger receptors on AMs were altered post-BMT, with loss of macrophage receptor with collagenous structure (MARCO), a critical receptor for recognition of P. aeurignosa. We determined that PGE2 signaled via elevated E prostanoid 2 (EP2) receptors on these AMs to inhibit their functions. Importantly, our murine studies have shown that pharmacologic or genetic inhibition of COX-2 post-BMT restores lung innate immunity and AM function against P. aeruginosa. These results are exciting because they suggest inhibition of PGE2 signaling can be a therapeutic to improve host defense post-transplant. However, there are systemic problems with a therapeutic strategy that globally blocks all prostaglandin synthesis. Thus, one aspect of this proposal will be to test a newly developed EP2 antagonist (PF-044148948) which we have obtained from Pfizer. We believe this will be a much more specific and effective therapeutic to block the inhibitory PGE2 signaling. The application also seeks to provide insight into the following unanswered questions. 1) Why is COX-2 elevated post-BMT leading to overproduction of PGE2? 2) Do these same innate immune defects characterize allogeneic (allo) BMT? 3) Do these defects post-BMT make mice more susceptible to Gram positive infections (like Streptococcus pneumoniae) as well as Gram negative ones? 4) Can we determine whether impairment of autophagy is one mechanism for impaired killing post-BMT? 5) Are the defects we have noted in our murine model also present in human HSCT patients? Our overall hypothesis is: BMT conditioning induces transforming growth factor (TGF) secretion from lung epithelial cells. This augments miR-29b expression to block synthesis of DNA methyltransferases (DNMTs) causing hypomethylation of COX-2 leading to PGE2 overexpression in AMs. Furthermore, PGE2-EP2 induced alterations in autophagy impair host defense against P. aeruginosa and S. pneumoniae post-BMT and we speculate that host defense post-BMT can be improved via treatment with a COX inhibitor, an EP2 antagonist or via induction of autophagy. These hypotheses will be explored in the following specific aims.
Aim 1) To determine if syn BMT and allo BMT mice are more susceptible to P. aeruginosa and S. pneumoniae infection and if susceptibility is related to PGE2 signaling via EP2 in mice and man Aim 2: To determine whether TGF-induced miR-29b causes COX-2 hypomethylation to increase PGE2 production post-BMT Aim 3:
Aim 3 : To determine if autophagy is impaired in syn and allo BMT AMs, to determine whether this is related to PGE2- EP2 signaling and the importance of autophagy to host defense post-BMT

Public Health Relevance

Hematopoietic stem cell transplantation as a cure for genetic and malignant diseases is hampered by the high incidence of infection (particularly in the lung) which occurs post- transplant. This project seeks to understand alterations in innate immune cells which occur as a result of the transplantation regimen. We provide mechanistic insight into why these changes occur, and how they limit the ability of lung macrophages to fight off bacterial infection post- transplant. Finally, we propose to test novel therapeutic strategies to improve host defense against infection post-transplantation. This work could significantly enhance the efficacy and safety of this therapy for many patients.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
5R01AI117229-03
Application #
9189677
Study Section
Lung Cellular, Molecular, and Immunobiology Study Section (LCMI)
Program Officer
Palker, Thomas J
Project Start
Project End
Budget Start
Budget End
Support Year
3
Fiscal Year
2017
Total Cost
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
073133571
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
Speth, Jennifer M; Bourdonnay, Emilie; Penke, Loka Raghu Kumar et al. (2016) Alveolar Epithelial Cell-Derived Prostaglandin E2 Serves as a Request Signal for Macrophage Secretion of Suppressor of Cytokine Signaling 3 during Innate Inflammation. J Immunol 196:5112-20
Podsiad, Amy; Standiford, Theodore J; Ballinger, Megan N et al. (2016) MicroRNA-155 regulates host immune response to postviral bacterial pneumonia via IL-23/IL-17 pathway. Am J Physiol Lung Cell Mol Physiol 310:L465-75
Gurczynski, Stephen J; Procario, Megan C; O'Dwyer, David N et al. (2016) Loss of CCR2 signaling alters leukocyte recruitment and exacerbates γ-herpesvirus-induced pneumonitis and fibrosis following bone marrow transplantation. Am J Physiol Lung Cell Mol Physiol 311:L611-27
Zhou, X; Loomis-King, H; Gurczynski, S J et al. (2016) Bone marrow transplantation alters lung antigen-presenting cells to promote TH17 response and the development of pneumonitis and fibrosis following gammaherpesvirus infection. Mucosal Immunol 9:610-20
O'Dwyer, David N; Ashley, Shanna L; Moore, Bethany B (2016) Influences of innate immunity, autophagy, and fibroblast activation in the pathogenesis of lung fibrosis. Am J Physiol Lung Cell Mol Physiol 311:L590-601
O'Dwyer, David N; Dickson, Robert P; Moore, Bethany B (2016) The Lung Microbiome, Immunity, and the Pathogenesis of Chronic Lung Disease. J Immunol 196:4839-47
Domingo-Gonzalez, Racquel; Martínez-Colón, Giovanny J; Smith, Alana J et al. (2016) Inhibition of Neutrophil Extracellular Trap Formation after Stem Cell Transplant by Prostaglandin E2. Am J Respir Crit Care Med 193:186-97
Moore, Bethany B; Moore, Thomas A (2015) Viruses in Idiopathic Pulmonary Fibrosis. Etiology and Exacerbation. Ann Am Thorac Soc 12 Suppl 2:S186-92
Russkamp, Norman F; Ruemmler, Robert; Roewe, Julian et al. (2015) Experimental design of complement component 5a-induced acute lung injury (C5a-ALI): a role of CC-chemokine receptor type 5 during immune activation by anaphylatoxin. FASEB J 29:3762-72
O'Flaherty, Brigid M; Matar, Caline G; Wakeman, Brian S et al. (2015) CD8+ T Cell Response to Gammaherpesvirus Infection Mediates Inflammation and Fibrosis in Interferon Gamma Receptor-Deficient Mice. PLoS One 10:e0135719

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