This Program Project competing renewal application has as its long-term objective the delineation of molecular events involved in macrophage activation for tumor cell killing. The research proposed will focus on gene activation and products of genes that collectively regulate the development and suppression of nitric oxide-dependent tumoricidal activity, in some cases through autocrine feedback. Early, intermediate and late gene expression will be studied. The three research projects are tightly integrated, both scientifically and fiscally. Project #1, led by David C. Morrison, will investigate molecular mechanisms of LPS- initiated signalling that lead to the development of activation and the expression of tumoricidal activity. He will have three specific aims: (i) to characterize the specific contributions of various LPS receptor molecules (e.g., p73, CD14, and CD11/18) to signal transduction, gene transcription, and activation for tumor cell killing; (ii) to define the molecular basis for LPS-mediated reprogramming of macrophages for altered function; and (iii) to explore the hypothesis that selective macrophage reprogramming also occurs in vivo in a model tumor system that renders mice hypersusceptible to the lethal effect of LPS. Project #2, directed by Tsuneo Suzuki, will investigate the sequence of cytoplasmic activation events that are initiated by LPS, some of which are dependent on auto/paracrine feedback of macrophage gene products.
Two specific aims will be pursued: (i) to investigate LPS-triggered signal transduction mechanisms that lead to the activation of NF-kappa-B, and (ii) to investigate whether or not macrophages can be activated in vitro and in vivo by tumor target cells that have been stably transfected or transduced with the genes encoding for either IFN-beta, IFN-gamma, monocyte chemotactic peptide (MCP-1, aka JE), or combinations thereof inserted into eukaryotic expression vectors. Project #3, led by Stephen W. Russell, will investigate events in the cell nucleus, specifically how LPS/IFN-mediated expresion of the gene that encodes for inducible nitric oxide synthase (iNOS) is regulated. There will be two specific aims in this project: (i) to identify the components of the network of transcription factors and responsive elements that positively/ negatively regulate mouse iNOS gene expression, and (ii) to identify the mechanism(s) by which IFN-beta affects iNOS production, both positively and negatively. The Support Services Core Component, which will also be led by Dr. Russell, is designed to facilitate, coordinate and foster research that is conducted by each of the three projects. It will meet three specific needs, namely those for (i) program coordination and facilitation, (ii) quality control and standardization of reagents and cells, and (iii) clerical support. Overall, the highly integrated approach that is proposed, coupled with a Core Component that will proactively foster research productivity, is expected to lead to extensive collaborations between the component projects and, therefore, to even greater productivity than has characterized the current performance period (34 full length manuscripts either published or submitted in 2+ years).

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Program Projects (P01)
Project #
5P01CA054474-06
Application #
2007930
Study Section
Special Emphasis Panel (SRC (CC))
Project Start
1992-02-01
Project End
1998-11-30
Budget Start
1996-12-20
Budget End
1997-11-30
Support Year
6
Fiscal Year
1997
Total Cost
Indirect Cost
Name
University of Kansas
Department
Miscellaneous
Type
Schools of Medicine
DUNS #
016060860
City
Kansas City
State
KS
Country
United States
Zip Code
66160
Zhang, Yue H; Murphy, William J; Russell, Stephen W et al. (2005) Serum-dependent potentiation of lipopolysaccharide-induced nitric oxide production is mediated by the events after the transcription of inducible type of nitric oxide synthase. Cell Immunol 234:16-22
Crespo, A; Filla, M B; Russell, S W et al. (2000) Indirect induction of suppressor of cytokine signalling-1 in macrophages stimulated with bacterial lipopolysaccharide: partial role of autocrine/paracrine interferon-alpha/beta. Biochem J 349:99-104
Gao, J J; Filla, M B; Lorsbach, R B et al. (2000) Prolonged exposure of mouse macrophages to IFN-beta suppresses transcription of the inducible nitric oxide synthase gene: altered availability of transcription factor Stat1alpha. Eur J Immunol 30:1551-61
Xia, D; Wang, F; Parmely, M J (2000) Inhibition of nuclear factor-kappab activation in mouse macrophages and the RAW 264.7 cell line by a synthetic adenyl carbocyclic nucleoside. Biochem Pharmacol 60:717-27
David, S A; Awasthi, S K; Balaram, P (2000) The role of polar and facial amphipathic character in determining lipopolysaccharide-binding properties in synthetic cationic peptides. J Endotoxin Res 6:249-56
David, S A; Silverstein, R; Amura, C R et al. (1999) Lipopolyamines: novel antiendotoxin compounds that reduce mortality in experimental sepsis caused by gram-negative bacteria. Antimicrob Agents Chemother 43:912-9
Morrison, D C; Silverstein, R; Luchi, M et al. (1999) Structure-function relationships of bacterial endotoxins. Contribution to microbial sepsis. Infect Dis Clin North Am 13:313-40
Gao, J J; Zuvanich, E G; Xue, Q et al. (1999) Cutting edge: bacterial DNA and LPS act in synergy in inducing nitric oxide production in RAW 264.7 macrophages. J Immunol 163:4095-9
Kielian, T; Nagai, E; Ikubo, A et al. (1999) Granulocyte/macrophage-colony-stimulating factor released by adenovirally transduced CT26 cells leads to the local expression of macrophage inflammatory protein 1alpha and accumulation of dendritic cells at vaccination sites in vivo. Cancer Immunol Immunother 48:123-31
Shnyra, A; Brewington, R; Alipio, A et al. (1998) Reprogramming of lipopolysaccharide-primed macrophages is controlled by a counterbalanced production of IL-10 and IL-12. J Immunol 160:3729-36

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