Orthotopic liver transplantation (OLT) is an effective therapeutic modality for the treatment of end-stage liver disease. However, ischemia/reperfusion injury (IRI), an exogenous Ag-independent component of the "harvesting" insult, remains one of the key limitations after OLT. Moreover, the quality of donor organs decreases along with the aging of general population and associated pathological conditions. This proposal is built upon the insights gained from our previous studies on a novel approach of combating organ IRI by locally inducing heme oxygenase-1 (HO-1) that protects against the severity of oxidative stress. Indeed, local HO-1 expression and functional response should be considered as a denominator of donor organ quality. We have also shown the role of innate immunity in liver IRI. Indeed, by utilizing a non-transplant mouse warm liver IRI model, we have documented that: (i) TLR4 activation mediates liver inflammation via IRF-3 pathway;(ii) CXCL10 regulates liver innate immune responses;(iii) Type-I IFN mediates synergy between Kupffer cells and hepatocytes, and (iv) endogenous TLR ligands are crucial in TLR4 activation-induced "sensoring" and IRI. To mimic the clinical scenario, we have recently developed a mouse model of prolonged liver cold preservation, followed by syngeneic OLT. We hypothesize that cross talk between the opposing pathways, i.e., HO-1 in the donor organ, and TLR4 in OLT recipients is instrumental in the mechanism of liver IRI. Our corollary hypothesis states that Nrf2, a bZIP transcriptional factor that regulates stress response/regulates cell redox balance, controls dysregulated HO-1 - TLR4 signaling during IRI.
Aim 1. To analyze mechanisms by which suboptimal HO-1 deficient liver grafts affect IRI sequel and TLR4 signaling in OLT recipients. Livers from WT, HO-1 deficient (+/-;KO) and HO-1 overexpressing (Tg) donors will be stored for 18 h at 4 C, and then transplanted to syngeneic WT mice. We will study (i) whether HO-1 expression and by which donor liver cell type affect IRI and OLT outcome, and (ii) if and how the local HO-1 expression in OLT affects host TLR4 signaling.
Aim 2. To dissect mechanisms by which modulation of recipient TLR4 signaling ameliorate IRI and improve the outcome of suboptimal OLTs. Livers from WT or HO-1 deficient (+/-;KO) donors will be stored for 18 h at 4 C, and then transplanted to syngeneic TRIF- or Type I IFN receptor (IFNAR)-KO mice. We will study (i) whether selective ablation of downstream TLR4 signaling in the host affect IRI sequel/OLT inflammation, and (ii) if HO-1- TLR4 cross talk can influence HMGB1-mediated IR inflammation and organ damage in OLT recipients.
Aim 3. To analyze mechanisms by which Nrf2 signaling influence IRI in OLT recipients. Livers from donor mice that are Nrf2-deficient or Nrf2-overexpressing (hepatocyte-specific conditional disruption of the Keap1 gene, which represses Nrf2) will be stored for 18 h at 4 C, and transplanted to WT mice. By modulating oxidant (HO-1 siRNA) and inflammatory (anti-HMGB1/rHMGB1) responses, we will study the regulatory function of Nrf2 upon (i) HO-1 vs (ii) TLR4 signaling pathways during IRI in OLT recipients.

Public Health Relevance

Host sensitization remains the major problem in clinical organ transplantation. Many prospective transplant patients are sensitized following blood transfusions, pregnancies, or failed previous grafts. This project is designed to analyze cell mediated mechanisms leading to accelerated rejection of organ allografts and ultimately to design novel and much needed therapeutic approaches to ameliorate transplant rejection in sensitized patients.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK062357-10
Application #
8432031
Study Section
Surgery, Anesthesiology and Trauma Study Section (SAT)
Program Officer
Doo, Edward
Project Start
2002-07-01
Project End
2014-01-31
Budget Start
2013-03-01
Budget End
2014-01-31
Support Year
10
Fiscal Year
2013
Total Cost
$316,795
Indirect Cost
$111,084
Name
University of California Los Angeles
Department
Surgery
Type
Schools of Medicine
DUNS #
092530369
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Huang, Jing; Yue, Shi; Ke, Bibo et al. (2014) Nuclear factor erythroid 2-related factor 2 regulates toll-like receptor 4 innate responses in mouse liver ischemia-reperfusion injury through Akt-forkhead box protein O1 signaling network. Transplantation 98:721-8
Huang, Jing; Shen, Xiu-Da; Yue, Shi et al. (2014) Adoptive transfer of heme oxygenase-1 (HO-1)-modified macrophages rescues the nuclear factor erythroid 2-related factor (Nrf2) antiinflammatory phenotype in liver ischemia/reperfusion injury. Mol Med 20:448-55
Ji, Haofeng; Liu, Yuanxing; Zhang, Yu et al. (2014) T-cell immunoglobulin and mucin domain 4 (TIM-4) signaling in innate immune-mediated liver ischemia-reperfusion injury. Hepatology 60:2052-64
Ke, Bibo; Shen, Xiu-Da; Zhang, Yu et al. (2013) KEAP1-NRF2 complex in ischemia-induced hepatocellular damage of mouse liver transplants. J Hepatol 59:1200-7
Kamo, Naoko; Ke, Bibo; Busuttil, Ronald W et al. (2013) PTEN-mediated Akt/*-catenin/Foxo1 signaling regulates innate immune responses in mouse liver ischemia/reperfusion injury. Hepatology 57:289-98
Zhang, Y; Ji, H; Shen, X et al. (2013) Targeting TIM-1 on CD4 T cells depresses macrophage activation and overcomes ischemia-reperfusion injury in mouse orthotopic liver transplantation. Am J Transplant 13:56-66
Kamo, Naoko; Ke, Bibo; Ghaffari, Amir A et al. (2013) ASC/caspase-1/IL-1* signaling triggers inflammatory responses by promoting HMGB1 induction in liver ischemia/reperfusion injury. Hepatology 58:351-62
Ji, Haofeng; Zhang, Yu; Shen, Xiu-da et al. (2013) Neuropeptide PACAP in mouse liver ischemia and reperfusion injury: immunomodulation by the cAMP-PKA pathway. Hepatology 57:1225-37
Ji, Haofeng; Zhang, Yu; Liu, Yuanxing et al. (2013) Vasoactive intestinal peptide attenuates liver ischemia/reperfusion injury in mice via the cyclic adenosine monophosphate-protein kinase a pathway. Liver Transpl 19:945-56
Ke, Bibo; Shen, Xiu-Da; Kamo, Naoko et al. (2013) ýý-catenin regulates innate and adaptive immunity in mouse liver ischemia-reperfusion injury. Hepatology 57:1203-14

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