While the exact mechanisms of ischemia/reperfusion injury remain unclear, our laboratory and others have shown that inhibition of complement activation (e.g., sCR1 or C1 esterase inhibitor), depletion of complement components (e.g., cobra venom factor) and complement component (e.g., C3 and C4) deficient mice reveal an important role of complement in ischemia/reperfusion injury. In the last funding cycle, we showed that inhibition of mannose binding lectin (MBL) protected the ischemic gastrointestinal system from the primary ischemic event. In contrast, the pulmonary system, which takes a second hit from the primary ischemic gastrointestinal tract, was not protected. Complement activation in the lung was observed in C1qa- or MBL-deficient mice, but not in C2/factor B deficient (KO) mice. Addition of C2 restored the complement activation and tissue inflammation/injury in the C2/fB KO mice following gastrointestinal ischemia/reperfusion (GI/R). These data suggest that a C2 dependent, but C1q or MBL independent, process is responsible for the complement activation, inflammation and injury following GI/R. Preliminary data suggest that another lectin, ficolin-A is the initiating molecule responsible for complement activation in the lungs following GI/R. Recent publications also showed a C2/C4 bypass mechanism involving MBL may be responsible for complement activation via the alternative pathway. Preliminary studies demonstrate biological evidence of this bypass mechanism in vitro and several potential molecular mechanisms involving the MBL complex and the coagulation system. Studies by others show several different interactions with the coagulation system and complement. Using the FeCl3 model of arterial thrombogenesis, we observed that thrombogenesis was absent in MBL null mice, yet present in C2/fB null mice. We show that trauma patients demonstrate a link between complement activation and coagulopathy with a significant decrease in MBL levels observed early on. At least one of these interacting links involves the direct activation of the alternative pathway by the MBL complex, bypassing the need for C2 or C4. Our preliminary data demonstrate multiple interactions of lectin complexes with coagulation proteins and may explain why inhibition of complement has proven unsuccessful in clinical studies for CABG and CAD, warranting a better understanding of the complex interactions of complement with the coagulation system. In this competitive renewal, we will continue to investigate the molecular mechanisms involved in complement activation during oxidative stress of endothelial cells in vitro and in vivo and its interactions with the coagulation cascade. The general aim of this application is to characterize the molecular mechanisms governing lectin induced activation of coagulation following endothelial oxidative stress. Studies in this renewal will investigate the molecular mechanism of complement activation and interactions of complement with coagulation proteins. We will also be characterizing the potential use of a novel complement inhibitor that was recently discovered, cloned and expressed since our last submission, and is part of the lectin complex.

Public Health Relevance

Clinical inhibition of complement has failed for ischemia and reperfusion diseases (e.g., CAD and CABG), despite basic science studies showing protection in animal models. This grant will characterize the role of a novel lectin, ficolin-A, in the pulmonary injury observed following gastrointestinal ischemia, while also investigating the connections between the lectin complexes'ability to activate the coagulation system. The data demonstrate the importance of selecting the appropriate complement inhibitor for human disease because of the connectivity observed in these two cascade systems: complement and coagulation.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL056086-17
Application #
8588951
Study Section
Surgery, Anesthesiology and Trauma Study Section (SAT)
Program Officer
Schwartz, Lisa
Project Start
1996-05-01
Project End
2014-11-30
Budget Start
2013-12-01
Budget End
2014-11-30
Support Year
17
Fiscal Year
2014
Total Cost
$375,131
Indirect Cost
$150,131
Name
Brigham and Women's Hospital
Department
Type
DUNS #
030811269
City
Boston
State
MA
Country
United States
Zip Code
02115
Zou, Chenhui; La Bonte, Laura R; Pavlov, Vasile I et al. (2012) Murine hyperglycemic vasculopathy and cardiomyopathy: whole-genome gene expression analysis predicts cellular targets and regulatory networks influenced by mannose binding lectin. Front Immunol 3:
Pavlov, Vasile I; La Bonte, Laura R; Baldwin, William M et al. (2012) Absence of mannose-binding lectin prevents hyperglycemic cardiovascular complications. Am J Pathol 180:104-12
Sebastian, Becky M; Roychowdhury, Sanjoy; Tang, Hui et al. (2011) Identification of a cytochrome P4502E1/Bid/C1q-dependent axis mediating inflammation in adipose tissue after chronic ethanol feeding to mice. J Biol Chem 286:35989-97
Takahashi, Kazue; Chang, Wei-Chuan; Takahashi, Minoru et al. (2011) Mannose-binding lectin and its associated proteases (MASPs) mediate coagulation and its deficiency is a risk factor in developing complications from infection, including disseminated intravascular coagulation. Immunobiology 216:96-102
Michelow, Ian C; Dong, Mingdong; Mungall, Bruce A et al. (2010) A novel L-ficolin/mannose-binding lectin chimeric molecule with enhanced activity against Ebola virus. J Biol Chem 285:24729-39
Gorsuch, W Brian; Guikema, Benjamin J; Fritzinger, David C et al. (2009) Humanized cobra venom factor decreases myocardial ischemia-reperfusion injury. Mol Immunol 47:506-10
Hart, Melanie L; Henn, Martina; Kohler, David et al. (2008) Role of extracellular nucleotide phosphohydrolysis in intestinal ischemia-reperfusion injury. FASEB J 22:2784-97
Hart, Melanie L; Much, Chressen; Gorzolla, Iris C et al. (2008) Extracellular adenosine production by ecto-5'-nucleotidase protects during murine hepatic ischemic preconditioning. Gastroenterology 135:1739-1750.e3
Yager, Phoebe H; You, Zerong; Qin, Tao et al. (2008) Mannose binding lectin gene deficiency increases susceptibility to traumatic brain injury in mice. J Cereb Blood Flow Metab 28:1030-9
Csencsits, K; Burrell, B E; Lu, G et al. (2008) The classical complement pathway in transplantation: unanticipated protective effects of C1q and role in inductive antibody therapy. Am J Transplant 8:1622-30

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