Scientific Report 2003-2004 - Cleveland Clinic Lerner Research ...

THE BORDENLABORATORYRESEARCH ASSOCIATESMamta Chawla-Sarkar, Ph.D.Kevin Taylor, Ph.D .POSTDOCTORAL FELLOWSSoo-In Bae, Ph.D.Vadim Budagian, Ph.D.Yan-Fang Liu, Ph.D.Navneet Majail, M.D.Paul Masci, D.O.Fred Reu, M.D.Snehal Thakar, M.D.TECHNICAL ASSOCIATESRon Grane, B.S.Barbara Jacobs, M.S.COLLABORATORSDaniel Lindner, M.D., Ph.D. 1Ganes C. Sen, Ph.D. 2Robert H. Silverman, Ph.D. 3Bryan R.G. Williams, Ph.D. 3Taolin Yi, Ph.D. 31Taussig Cancer Center, CCF2Dept. of Molecular Biology,CCF3Dept. of Cancer Biology, CCF172Investigations of Interferon Add Insight forUse in Combined Modality Cancer TreatmentsInterferons (IFNs) have proven an importantparadigm for establishing the role ofbiologicals as effective therapy in humanmalignancies. Despite this substantial progress,we still do not understand the underlyingmechanisms of tumor sensitivity and resistance.How to overcome resistance to IFNs innonresponding patients has been little explored.It is our overall hypothesis that clinical resistanceresults from deficiencies in components of theJAK/STAT signal transduction pathway and/orexpression of specific interferon-stimulatedgenes (ISGs); conversely, tumor response will beaugmented by increases in signal transductionand/or expression of specific ISGs.IFNs have proven active as single agents insome tumors (melanoma, myeloma, renal cellcarcinoma, chronic leukemias, T- and B-celllymphomas, carcinoids, and Kaposi’s sarcoma)and are now being increasingly integrated intocombined modality therapy with clinicallybeneficial effects. IFNs are possibly the mostpotent modulators of gene expression used inclinical medicine. Since the clinical effects almostcertainly result from ISGs, to probe underlyingmechanisms of antitumor action, we havefocused on ISGs and their control. We havedemonstrated modulation of such genes and theirproducts in treated patients in vivo. These studieshave defined correlations between doses,schedule of administration, and cytokine typewith gene induction and cellular response.One goal is thus to define defects inexpression or activation of signal transductioncomponents and extend our studies of interventionsto correct abnormalities. We have identifiedantiestrogens, demethylating agents, phosphataseChawla-Sarkar, M., Leaman, D.W., and E.C. Borden (2001) Preferential induction ofapoptosis by interferon (IFN)-beta compared with IFN-alpha2: correlation withTRAIL/Apo2L induction in melanoma cell lines. Clin. Cancer Res. 7:1821-1831.Leaman, D.W., Chawla-Sarkar, M., Vyas, K., Reheman, M., Tamai, K., Toji, S., andE.C. Borden (2002) Identification of X-linked inhibitor of apoptosis-associated factor-1 as an interferon-stimulated gene that augments TRAIL Apo2L-induced apoptosis.J. Biol. Chem. 277:28504-28511.Chawla-Sarkar, M., Leaman, D.W., Jacobs, B.S., and E.C. Borden (2002) IFN-betapretreatment sensitizes human melanoma cells to TRAIL/Apo2 ligand-induced apoptosis.J. Immunol. 169:847-855.Padovan, E., Terracciano, L., Certa, U., Jacobs, B., Reschner, A., Bolli, M., Spagnoli,G.C., Borden, E.C., and M. Heberer (2002) Interferon stimulated gene 15 constitutivelyproduced by melanoma cells induces E-cadherin expression on humandendritic cells. Cancer Res. 62:3453-3458.Chawla-Sarkar, M., Leaman, D.W., Jacobs, B.S., Tuthill, R.J., Chatterjee-Kishore,M., Stark, G.R., and E.C. Borden (2002) Resistance to interferons in melanomacells does not correlate with the expression or activation of signal transducer andactivator of transcription 1 (Stat1). J. Interferon Cytokine Res. 22:603-613.Pathak, M.K., Dhawan, D., Lindner, D.J., Borden, E.C., Farver, C., and T. Yi(2002) Pentamidine is an inhibitor of PRL phosphatases with anticancer activity.Mol. Cancer Ther. 1:1255-64.inhibitors and cytokines as molecules thataugment IFN signal transduction and antitumoreffects in preclinical models. Such defects mayinfluence not only response of tumors to IFNsbut also to other cytokines that work via IFNassociatedsignal transduction pathways. Since,however, not all tumors will have signal transductionabnormalities, it seems likely that selectivelack of expression of specific ISGs will accountfor resistance in other tumors.A second goal is to identify specific ISGsthat are induced in human tumors in response toIFN-α2 and IFN-β. Through gene chip technologyin vitro, new genes have been identified thatmay be particularly critical in mediating theantitumor effects. Toward this end, we aredissecting the influence on cell function of thesegene products and are assessing newly identifiedISGs in patients receiving IFNs and otherbiological response modifiers. In collaborationwith Drs. Bryan Williams and Robert Silverman,our focus is particularly on newly identified ISGsthat influence cell proliferation and/or apoptosis.The pleiotropic effects of IFNs on cellfunction have led us to assess not onlyantiproliferative and apoptotic effects on tumorcells but also components of host response.Influences of IFNs on immune effector cellfunction that have been defined include effects onboth natural killer/lymphokine-activated killer(NK/LAK) cell activity and monocytes. We haveidentified a new lymphokine, ISG15 (itself anISG protein product), that influences NK/LAKcell function. Another important component ofhost-tumor interaction is endothelial cellproliferation. In collaboration with Dr. DanielLindner, we are continuing to assess the augmentedanti-angiogenic effects of IFNs incombination with other molecules in causingtumor inhibition. Finally, although apoptosis hasnot been considered a prominent part of themechanism of action of IFNs, our recent studiesdefine a caspase-mediated event induction in celllines of several histologies.The clinical and gene modulatory effects ofmost members of the family of IFN proteins andof IFN inducers have hardly begun to be assessed.Thus we are beginning to more extensivelyevaluate other members of the IFN multigenefamily in more depth. We are thus assisting, withDr. Silverman and Dr. Ganes Sen, in probing theantitumor effects of other molecules that harnessthe IFN system for potentially greater clinicaleffectiveness. We are also evaluating otherbiologicals or low-molecular-weight moleculesthat modify host response, such as other cytokines(interleukin [IL]-2, -6, -10, and -12), monoclonalantibodies, and tumor vaccines that havecomplementary mechanisms of cellular andantitumor effects of IFNs. It is our goal in all ofthese studies to translate important leads withIFNs and other biological modifiers into moreeffective therapies.