REGULATION OF GRANULOCYTE AND MACROPHAGE PRODUCTION

Colony Stimulating Factor-1 (CSF-1) regulates the survival, proliferation and differentiation of mononuclear phagocytes that appear to play critical roles in the development and function of the tissues in which they reside. In addition, CSF-1 is import in female fertility via its direct action on c ls of the female reproductive tract, which apart from mononuclear phagocytes, include decidual cells, trophoblast, oocytes and embryonic cells. Not surprisingly, therefore, CSF-1 appears to have important autocrine and/or paracrine roles in neoplasias of the myeloid, lymphoid and female reproductive systems. All of the effects of CSF-1 transduces its signals f survival, proliferation and differentiation of the mononuclear phagocyte. The specific aims are: 1. To identify novel components of CSF-1 signal transduction pathways. 2. To elucidate the functions of components of CSF-1 signal transduction pathways. 3. To develop novel systems for the study of CSF-1 receptor mediated signaling in mononuclear phagocytes. 4. To analyze to role to protein tyrosine phosphatase-phi in macrophage survival, proliferation and function. GRANT R01CA31611 The commonly eaten, cultivated mushroom, Agaricus bisporus (AB), contains several N-N-bond-containing chemicals. We demonstrated earlier the presence, carcinogenicity and mode of action of some of these mushroom chemicals. In the recent grant, we have shown the carcinogenicity of the uncooked mushroom and some of its ingredients in long-term experiments in mice. In addition, we have revealed the presence of two hydrazines: p-hydrazinobenzoic acid and beta-N-[gamma-L(=)- glutamyl]-4-carboxyphenylhydrazine in the mushroom extracts. Further, we demonstrated in vitro the metabolism of several mushroom arylhydraines using cytochrome P-450 mixed function oxidases and prostaglandin (H) synthase enzymes. Finally, using purine bases and nucleosides, we have modeled the reactions of 4- (hydroxymethyl)benzenediazonium ion (HMBD) with DNA and have observed that HMBD can react with DNA. Secondary reactions (e.g., cross-linking) may be important. Lastly, we determined the amount of hydrazines in the mushroom after baking: Only 25% was destroyed. Also, using the Ames assay we noted no significant difference between the mutagenic activities of the baked and unbaked mushroom extracts. In this new continuing proposal, we intend to extend our investigations to: (1) study the carcinogenicity of baked, lyophilized and raw AB by using a dose response protocol in lifelong experiments in mice; (2) reveal the tumorigenicity of 4-(hydroxy)benzenediazonium sulfate in mice; (3) analyze AB grown by us using radiolabeled compounds for the presence of diazonium ions and hydrazines; (4) determine the products of the reaction of HMBD, other diazonium ions and their reactive species with bases, nucleosides and DNA in vitro at physiological pH and in isolated cells; and (5) synthesize the above and other chemicals for analytical chemistry, biochemical and carcinogenicity studies. The annual estimated mushroom consumption in the United States was approximately 435 million kilograms in 1990-91. The corresponding figure in western Europe was more than 644 million kilograms. No new studies are proposed. The on-going baked and lyophilized AB mushroom experiments and the accompanying chemistry and biochemistry investigations are positive. Therefore, the projects have practical implications because humans consume large quantities of baked and lyophilized mushrooms. Further, the mechanistic studies provided interesting clues in reveling their modes of action on the molecular level. Another year of funding is requested for the completion of these studies.