Research in my laboratory is focused on the role of polyamines in cancer cell proliferation and ways to exploit their metabolism and function as antiproliferative and chemopreventive targets. Polyamines are naturally occurring polycationic alkylamines that are absolutely required for eukaryotic cell growth and differentiation. Their intracellular concentrations (millimolar) are maintained in a narrow range through the actions of a highly regulated and specific transport system and a rapidly responding metabolic pathway. The first rate-limiting step in the production of polyamines is the decarboxylation of ornithine by the highly inducible and short-lived enzyme, ornithine decarboxylase (ODC). Increased ODC activity and new polyamine synthesis have frequently been associated with the neoplastic phenotype. The catabolism of the polyamines is controlled by the rate limiting enzyme spermidine/spermine N1-acetyltransferase (SSAT) that my laboratory has cloned and continues to study. This enzyme is highly controlled at the level of transcription, translation and is stabilized by the natural polyamines and antitumor polyamine analogues. We have also demonstrated that the phenotype-specific antitumor activity of specific polyamine analogues is associated with a superinduction of this enzyme in response to treatment. Another enzyme cloned in my laboratory is the inducible FAD-dependent, spermine oxidase (SMO) that is also a polyamine catabolic enzyme that produces H2O2 as one of its products. SMO is induced by many inflammatory stimuli including bacterial infection and inflammatory cytokines. As an example, Helicobacter pylori, a causative agent of gastric cancer, highly induces SMO resulting in sufficient H2O2 production to produce oxidative DNA damage. Thus it appears that SMO may be one of the molecular links between inflammation and carcinogenesis and as such represents a potential target for chemopreventive intervention. During the studies that lead to the discovery of SMO, we also identified the sequence of a related FAD-dependent oxidase that was later identified as lysine specific demethylase 1 (LSD1). LSD1 is an important chromatin-remodeling enzyme that demethylates mono- and dimethyl lysine 4 of histone 3 (H3K4me1 & H3K4me2). As these histone marks are associated with active transcription, LSD1 has the ability to broadly repress gene transcription. LSD1 activity is involved with the inappropriate silencing of several tumor suppressor gene involved in the etiology and progression of cancer. As LSD1 is structurally and functionally homologous to SMO, we hypothesized that certain polyamine analogues would inhibit LSD1 and lead to the re-expression of inappropriately silenced genes. We have now demonstrated that this is, in fact, the case and are pursuing this strategy to develop agents that may be useful in the treatment of neoplastic disease. Consequently, polyamine metabolism and function, and related pathways present a target rich environment against which therapeutic and chemopreventive strategies may be developed.

polyamines, polyamine oxidase, SSAT, PAO, PAOh1, SMO, apoptosis, reactive oxygen species, LSD1, epigenetics, chromatin modification, lysine demethylation

Carew, J.S., Nawrocki, S.T., Reddy, V.K., Bush, D., Rehg, J.E., Goodwin, A., Houghton, J.A., Casero, R.A., Marton, L.J., and Cleveland, J.L. The Novel Polyamine Analogue CGC-11093 Enhances Antimyeloma Activity of Bortezomib. Cancer Res. 68: 4783-4790, 2008

Hacker, A., Marton, L.J., Sobolewski, M., Casero, R.A., Jr. In Vitro and In Vivo Effects of the Conformationally Restricted Polyamine Analogue CGC-11047 on Small Cell and Non-small Cell Lung Cancer Cells. Can Chemo and Pharm 63:45-53, 2008.

Murray-Stewart, T., Wang, Y., Goodwin, A., Hacker, A., Meeker, A., and Casero, R.A., Jr. Nuclear Localization of Human Spermine Oxidase Isoforms: Implications in Drug Response and Disease Etiology. FEBS J., 275:2795-2806, 2008.

Goodwin, A., Jadallah, S., Toubaji, A., Lecksell, K., Hicks, J.L., Kowalski, J., Bova, G.S., DeMarzo, A.M., Netto, G.J., Casero, R.A. Jr. Increased Spermine Oxidase Expression in Human Prostate Cancer and Prostatic Intraepithelial Tissues. Prostate 68:766-772, 2008.

Varghese, S., Senanayake, T., Murray Stewart, T., Doering, K., Fraser, A., Casero, R.A., Jr., Woster, P.M. Polyaminohydroxamic Acids and Polyaminobenzamides as Isoform Selective Histone Deaceylase Inhibitors. J Med Chem. 51:2447-2456, 2008.

Huang, Y., Greene, E., Murray-Stewart, T., Goodwin, A.C., Baylin, S.B., Woster, P.M., and Casero, R.A., Jr. Inhibition of the Lysine Specific Demethylase, LSD1, by Novel Polyamine Analogues Results in Re-expression of Abberantly Silenced Genes. Proc. Natl. Acad. Sci. USA In 104:8023-8028, 2007.

Babbar, N., Murray-Stewart, T., and Casero, R.A., Jr. Inflammation and Polyamine Catabolism-The good, the bad, and the ugly. Biochem. Soc. Trans. 35:300-304, 2007.

Casero, R. and Marton, L. Targeting Polyamine Metabolism and Function in Cancer and Other Hyperproliferative Diseases. Nature Rev. Drug Devel. 6:373-390, 2007.

Babbar, N., Murray-Stewart, T., and Casero, R.A., Jr. Inflammation and Polyamine Catabolism Biochem. Soc. Trans. 35:300-304, 2007.

Rider, J., Hacker, A., Mackintosh, A., Pegg, A.E., Woster, P.M., and Casero, R.A., Jr. Spermine and Spermidine Mediate Protection Against Oxidative Damage Caused by Hydrogen Peroxide. Amino Acids 33:231-240, 2007.

Jiang, R., Choi, W., Khan, A., Hess, K., Gerner, G., Casero, R., Yung, W.K., Hamilton, S.R., Zhang, W. Activation of Polyamine Catabolism by N1, N11-dietthynorspermine Leads to Cell Death in Glioblastoma. Int J Onc 314:431-40, 2007

Babbar, N. and Casero, R.A., Jr. Tumor Necrosis Factor a Increases Reactive Oxygen Species by Inducing Spermine Oxidase in Human Lung Eptithelial Cells: A Potential Mechanism for Inflammation-Induced Carcinogenesis. Cancer Res., 66:11125-11130, 2006.

Babbar, N., Hacker, A., Huang, Y., and Casero, R.A., Jr. Tumor Necrosis Factor a Induces Spermidine/Spermin N1-Acetyltranferase Through Nuclear Factor ?B in Non Small Cell Lung Cancer Cells. J. Biol. Chem., 394:317- 324, 2006.

Babbar, N., Gerner, E.W., and Casero, R.A., Jr. Induction of Spermidine/Spermine N1-Acetyltransferase (SSAT) by Aspirin in Caco-2 Colon Cancer Cells. Biochem. J. 394:317-324, 2006.

Pledgie, A., Huang, Y., Hacker, A., Zhang, Z., Woster, P.M., Davidson, N.E., Casero, R.A., Jr. Spermine Oxidase SMO(PAOhl), Not N'-Acetylpolyamine Oxidase (PAO) is the Primary Source of Cytotoxic H202 in Polyamine Analogue¬treated Human Breast Cancer Cell Lines. J Bio Chem. 280:39843-39851, 2005.

Shi, Y., Lan, F., Matson, C., Mulligan, P., Whetstine, J.R., Cole, P.A., Casero, R.A., and Shi, Y. Histone Demethylation Mediated by the Nuclear Amine Oxidase Homolog LSD1. Cell 119:941-953, 2004.