project

The Center for Cardiopulmonary Pharmacology, operating within the Department of Pharmacological Sciences of the University of Milan, has been involved in eicosanoid research since its formation, and more recently, under the impulse of its current Scientific Director (prof. Giancarlo Folco) has focused its attention on the transcellular biosynthesis of 5-lipoxygenase metab olites and, in particular, the cysteinyl leukotrienes.
Cysteinyl leukotrienes (Cys-LT) represent a prominent family of eicosanoids derived from arachidonic acid via the 5-lipoxygenase pathway (5-LO) and are considered important paracrine mediators of inflammation. Their biological effects are mediated via two distinct G-protein coupled receptors (Cys-LT1 and Cys-LT2), characterized by a different pharmacological profile and tissue expression. The synthesis of Cys-LT is a stepwise event that requires the formation of an unstable intermediate (LTA4) and occurs according to a strict cellular specificity; however Cys-LT can also be generated through cellular cooperation mechanisms whereby cells (e.g. neutrophils) that possess 5-LO (the primary enzyme) synthesize LTA4 which is then exported to acceptor cells (e.g. platelets or endothelial cells), devoid of 5-LO, for further metabolism to Cys-LT via a LTC-4 synthase or a GST-II enzyme (the secondary enzymes). This process, termed transcellular biosynthesis (1), could be of special relevance in diseases such as cardiac infarction (2) or atherosclerosis characterized by documented inflammatory conditions (3).

a) Cys-LT receptor antagonists; b) Cys-LT synthetase inhibitors; c) pharmacological interventions to facilitate (or block) the interactions between circulating PMN and the vessel wall; e) use of transgenic animal models (5).
This research project intends to evaluate the role of Cys-LT formed and acting "in situ", in the cerebral vasculature, as a consequence of the cooperation between circulating PMN and the vessel intima, and faces the problem from molecular aspects to the complexity and intricacy of the "in vivo" situation.
We hope that it will provide support for novel therapeutic applications targeting affections of the cerebral vasculature.

References

1. Maclouf J, Folco G, Patrono C. Eicosanoids and iso-eicosanoids: constitutive, inducible and transcellular biosynthesis in vascular disease. Thromb Haemost, 1998, 79:691-705.

2. Folco G, Rossoni G, Buccellati C, Berti F, Maclouf J, Sala A. Leukotrienes in cardiovascular diseases. Am J Respir Crit Care Med, 2000, 161:S112-116.

3. Mehrabian M, Allayee H, Wong J, Shi W, Wang XP, Shaposhnik Z, Funk CD, Lusis AJ, Shih W. Identification of 5-lipoxygenase as a major gene contributing to atherosclerosis susceptibility in mice. Circ Res. 2002 Jul 26;91(2):120-6.

4. Librizzi L, Folco G, de Curtis M. Nitric oxide synthase inhibitors unmask acetylcholine-mediated constriction of cerebral vessels in the in vitro isolated guinea-pig brain. Neuroscience, 2000, 101:283-287.

5) Beller TC, Maekawa A, Friend DS, Austen KF, Kanaoka Y. Targeted gene disruption reveals the role of the cysteinyl leukotriene 2 receptor in increased vascular permeability and in bleomycin-induced pulmonary fibrosis in mice. J Biol Chem. 2004 Oct 29;279(44):46129-34. Epub 2004 Aug 24.

Recent Publications

1. Profita M, Sala A, Bonanno A, Riccobono L, Siena L, Melis MR, Di Giorgi R, Mirabella F, Gjomarkaj M, Bonsignore G, Vignola AM. Increased prostaglandin E2 concentrations and cyclooxygenase-2 expression in asthmatic subjects with sputum eosinophilia. Journal of Allergy and Clinical Immunology, 2003, 112(4): 709-716

2. Tedeschi A, Ciceri P, Zarini S, Lorini M, Di Donato M, Nicosia S, Miadonna A, Sala A. Role of sodium in intracellular calcium elevation and leukotriene B4 formation by agonist activated human neutrophils. Biochemical Pharmacology, 2004, 67(2):385-393

3. Di Gennaro A, Carnini C, Buccellati C, Ballerio R, Zarini S, Fumagalli F, Viappiani S, Librizzi L, Hernandez A, Murphy RC, Constantin G, de Curtis M, Folco G, Sala A. Cysteinyl-leukotriene receptors activation in brain inflammatory reactions and cerebral edema formation: a role for transcellular biosynthesis of cysteinyl-leukotrienes. FASEB J, 2004, 18:842-844

4. Moretti RM, Montagnani Marelli M, Sala A, Motta M, Limonta P. Activation of the orphan nuclear receptor ror? counteracts the proliferative effect of fatty acids on prostate cancer cells: crucial role of 5-lipoxygenase. International Journal of Cancer, 2004, 112:87-93

5. Santus P, Sola A, Carlucci P, Fumagalli F, Di Gennaro A, Mondoni M, Carnini C, Centanni S, Sala A. Lipid peroxidation and 5-lipoxygenase activity in Chronic Obstructive Pulmonary Disease subjects. American Journal of Respiratory and Critical Care Medicine, 2005, 171:838-843

6. Profita M, Di Giorgi R, Sala A, Bonanno A, Riccobono L, Mirabella F,Gjomarkaj M, Bonsignore G, Bousquet J, Vignola AM. Muscarinic receptors, leukotriene B4 production and neutrophilic inflammation in COPD patients. Allergy, 2005, 60(11):1361-1369

7. Coppola A, D'Angelo A, Fermo I, Mazzola G, Di Minno MND, Cajani A, Sala A, Folco G, Tremoli E, Di Minno G. Reduced in vivo oxidative stress following 5-methyltetrahydrofolate supplementation in patients with early onset thrombosis and 677TT methylenetetrahydrofolate reductase genotype. British Journal of Haematology, 2005, :1-9

8. Bolego C, Buccellati C, Radaelli T, Cetin I, Puglisi L, Folco G, Sala A. eNOS, COX-2 and prostacyclin production are impaired in endothelial cells from diabetics. Biochemical Biophysical Research Communications, 2005, in press