2004 Scientific Report

Jorge Pisarello,

Jorge Pisarello, Spectrum Health, Grand Rapids, Michigan Anthony Schaeffer and John Grayhack, Northwestern University, Evanston, Illinois Peter Schirmacher, University of Cologne, Germany Bin S. Teh and E. Brian Butler, Baylor College of Medicine, Houston, Texas Cornelius Verweij, University of Amsterdam, The Netherlands Recent Publications Qui, J., J. Madoz-Gurpide, D.E. Misek, R. Kuick, D.E. Brenner, G. Michailidis, B.B. Haab, G.S. Omenn, and S.M. Hanash. In press. Development of natural protein microarrays for diagnosing cancer based on an antibody response to tumor antigens. Journal of Proteome Research. Zhou, H., K. Bouwman, M. Schotanus, C. Verweij, J.A. Marrero, D. Dillon, J. Costa, P. Lizardi, and B.B. Haab. In press. Two-color, rolling circle amplification on antibody microarrays for sensitive, multiplexed serum-protein measurements. Genome Biology. Haab, B.B., and H. Zhou. 2004. Multiplexed protein analysis using spotted antibody microarrays. Methods in Molecular Biology 278: 33–46. Bouwman, Kerri, Ji Qiu, Heping Zhou, Mark Schotanus, Leslie A. Mangold, Robert Vogt, Erik Erlandson, John Trenkle, Alan W. Partin, David Misek, Gilbert S. Omenn, Brian B. Haab, and Samir Hanash. 2003. Microarrays of tumor cell–derived proteins uncover a distinct pattern of prostate cancer serum immunoreactivity. Proteomics 3(11): 2200–2207. Haab, Brian B. 2003. Methods and applications of antibody microarrays in cancer research. Proteomics 3(11): 2116–2122. Miller, Jeremy C., Heping Zhou, Joshua Kwekel, Robert Cavallo, Jocelyn Burke, E. Brian Butler, Bin S. Teh, and Brian B. Haab. 2003. Antibody microarray profiling of human prostate cancer sera: antibody screening and identification of potential biomarkers. Proteomics 3(1): 56–63. Furge, Kyle A., Ramsi Haddad, Jeremy Miller, Brian B. Haab, Jacqueline Schoumans, Bin T. Teh, Lonson L. Barr, and Craig P. Webb. 2002. Genomic profiling and cDNA microarray analysis of human colon adenocarcinoma and associated peritoneal metastases reveals consistent cytogenetic and transcriptional aberrations associated with progression of multiple metastases. Applied Genomics and Proteomics 1(2): 123–134. Guo, X., W.-O. Lui, C.-N. Qian, J.D. Chen, S.G. Gray, D. Rhodes, B. Haab, E. Stanbridge, H. Wang, M.-H. Hong, H.-Q. Min, C. Larsson, and B.T. Teh. 2002. Identifying cancer-related genes in nasopharyngeal carcinoma cell lines using DNA and mRNA expression profiling analyses. International Journal of Oncology 21(6): 1197–1204. From left to right: Shanmugam, Nikkel, Haab, Orchekowski, Hamelinck, Forrester 32

Laboratory of Cancer Pharmacogenetics Han-Mo Koo, Ph.D. Dr. Koo received his Ph.D. in microbiology and molecular genetics at Rutgers–The State University of New Jersey in 1993. In June 1999, he joined VARI as a Scientific Investigator. Dr. Koo passed away after a battle with cancer in May 2004, as this Report was going to press. Staff Paula Davidson, M.S. Matt VanBrocklin, M.S. Susan Kitchen, B.S. Laboratory Members Students Stephanie Ellison, B.S. Research Interests Advances in our understanding of the molecular pathophysiology of human cancers open promising opportunities for the prevention of and intervention in cancer. Our laboratory is interested in studying mechanisms of drug actions, identifying novel therapeutic targets, and developing novel anticancer agents by means of molecular-targeting approaches. Activating mutations in RAS oncogenes are the most frequent gain-of-function mutations detected in human cancers. Besides their welldocumented role in cellular transformation and tumorigenesis, we have previously shown that the RAS oncogenes play an important role in sensitizing tumor cells to deoxycytidine analogues such as 1-(β-D-arabinofuranosyl)cytosine (Ara-C) and gemcitabine, as well as topoisomerase (topo) II inhibitors, namely etoposide. These results are supported by clinical findings that patients who have RAS oncogene–positive acute myeloid leukemia show an increased remission rate, longer remission duration, and improved overall survival in response to a combination therapy of Ara-C plus topo II inhibitor. To translate our results into a clinical trial, we have established a collaboration with the Grand Rapids Clinical Oncology Program and the Spectrum Health Cancer Program. We have initiated a Phase II clinical trial to evaluate gemcitabine/etoposide combination treatment for patients with locally advanced or metastatic pancreatic carcinomas, over 95% of which display RAS oncogene activation. This trial was initiated in October 2002, and currently 19 patients are enrolled. Although not truly indicative, serum protein CA 19.9 is currently the best tumor marker for this and other GI cancers. Serum samples have been collected prior to and throughout the study for further proteomic analysis, with an emphasis on uncovering a better tumor marker, as well as developing novel therapeutic targets. Mitogen-activated protein kinase (MAPK) signaling pathways are highly conserved among all eukaryotes and are integral for the transduction of a variety of extracellular signals. Furthermore, constitutive activation of MAPK signaling (e.g., the Raf-MEK1/2-ERK1/2 pathway) contributes to many aspects of human cancers; hence, the pathway has been identified as a potential target for cancer intervention. Typically, cancer cells exhibit a cytostatic (growth arrest) response to the disruption of MAPK signaling. However, we have recently demonstrated that interfering with the MAPK signaling pathway evokes a cytotoxic response (apoptosis) in human melanoma cells but not in normal melanocytes. Both anthrax lethal toxin (which proteolytically cleaves MAPK kinases [MEKs]) and small-molecule MEK inhibitors (such as PD90859 and U0126) can trigger an apoptotic response in human melanoma cells. Normal melanocytes treated with the same inhibitors, on the other hand, simply arrest in the G1 phase of the cell cycle. More importantly, in vivo treatment with anthrax lethal toxin of human melanoma xenograft tumors in athymic nude mice provides either significant or complete tumor regression without apparent side effects. These results indicate that the MAPK signaling pathway represents tumor-specific survival signaling in melanoma and that inhibition of this pathway may be a useful and potentially selective strategy for treating this cancer. 33