11 months ago

2005 Scientific Report

  • Text
  • Report
  • Institute
  • Mice
  • Proteins
  • Signaling
  • Protein
  • Michigan
  • Tumor
  • Molecular
  • Laboratory

Jose Costa and Paul

Jose Costa and Paul Lizardi, Yale University School of Medicine, New Haven, Connecticut Deborah Dillon, Brigham and Women’s Hospital, Boston, Massachusetts Ziding Feng and Samir Hanash, Fred Hutchinson Cancer Research Center, Seattle, Washington Jorge Marrero, University of Michigan Hospital, Ann Arbor Alan Partin, Johns Hopkins University, Baltimore, Maryland Peter Schirmacher, University of Cologne, Germany Robert Vessella, University of Washington, Seattle Cornelius Verweij, University of Amsterdam, The Netherlands Recent Publications Hamelinck, D., H. Zhou, L. Li, Z. Feng, C. Verweij, D. Dillon, J. Costa, and B.B. Haab. In press. “Optimized normalization for antibody microarrays and the identification of serum protein alterations associated with pancreatic cancer.” Molecular & Cellular Proteomics. Haab, B.B., and P.M. Lizardi. In press. “RCA-enhanced protein detection arrays.” Methods in Molecular Biology. Breuhahn, Kai, Sebastian Vreden, Ramsi Haddad, Susanne Beckebaum, Dirk Stippel, Peer Flemming, Tanja Nussbaum, Wolfgang H. Caselmann, Brian B. Haab, and Peter Schirmacher. 2004. Molecular profiling of human hepatocellular carcinoma defines mutually exclusive interferon regulation and insulin-like growth factor II overexpression. Cancer Research 64(17): 6058–6064. Konwinski, R., R. Haddad, J.A. Chun, S. Klenow, S. Larson, B.B. Haab, and L.L. Furge. 2004. Oltipraz, 3H-1,2-dithiole-3-thione and sulforaphane induce overlapping and protective antioxidant responses in murine microglial cells. Toxicology Letters 153(3): 343–355. Lindvall, Charlotta, Kyle Furge, Magnus Björkholm, Xiang Guo, Brian Haab, Elisabeth Blennow, Magnus Nordenskjöld, and Bin Tean Teh. 2004. Combined genetic and transcriptional profiling of acute myeloid leukemia with normal and complex karyotypes. Haematologica 89(9): 1072–1081. Qiu, Ji, Juan Madoz-Gurpide, David E. Misek, Rork Kuick, Dean E. Brenner, George Michailidis, Brian B. Haab, Gilbert S. Omenn, and Sam Hanash. 2004. Development of natural protein microarrays for diagnosing cancer based on an antibody response to tumor antigens. Journal of Proteome Research 3(2): 261–267. Zhou, Heping, Kerri Bouwman, Mark Schotanus, Cornelius Verweij, Jorge A. Marrero, Deborah Dillon, Jose Costa, Paul Lizaardi, and Brian B. Haab. 2004. Two-color, rolling-circle amplification on antibody microarrays for sensitive, multiplexed serum-protein measurements. Genome Biology 5(4): R28. From left to right: Haab, Hamelinck, Orchekowski, Chen, Shafer, Forrester 30

Molecular Medicine and Virology Group Sheri L. Holmen, Ph.D. Dr. Holmen received her M.S. in biomedical science from Western Michigan University in 1995 and her Ph.D. in tumor biology from the Mayo Clinic College of Medicine in 2000. She did her postdoctoral work at VARI in the laboratory of Bart Williams from 2000 to 2003 and became a Junior Investigator at VARI in December 2003. Laboratory Members Staff Marleah Russo Research Interests T he primary focus of the Molecular Medicine and Virology Group is to identify molecules that can be effective targets for cancer therapy, with the goal of developing better therapies with fewer side effects. The sequencing of the human genome has yielded a wealth of biological data, but we still know relatively little about which genes are causally associated with tumor development and which are only markers of cancer. Because of the high cost of developing new therapies, it is important that we identify which genetic changes can be productively targeted. We are concentrating our initial efforts on melanoma and glioblastoma, tumors which demonstrate constitutive activation of Ras signaling. The RCAS system We use a series of replication-competent retroviral vectors based on the SR-A strain of Rous sarcoma virus (RSV), a member of the avian leukosis virus (ALV) family, to study the roles of different genes in tumor initiation and progression. RSV is the only known naturally occurring, replication-competent retrovirus that carries an oncogene, src. In the RCAS vectors, the region encoding src (which is dispensable for viral replication) has been replaced by a synthetic DNA linker. Foreign genes inserted into this linker are expressed from the viral LTR promoter via a subgenomic splice site (just as src is in RSV). RCAN vectors differ from RCAS vectors in that they lack the src splice acceptor; the gene of interest is inserted along with an internal promoter. Higher-titer viruses subsequently have been generated by replacing the RSV SR-A pol gene with the pol gene of the Bryan strain of RSV. These vectors are termed RCASBP or RCANBP. The ability of these vectors to infect non-avian cells relies on expression of the corresponding receptor on the cell surface. The viral receptor is typically introduced into mammalian cells (or mice) via an inducible and/or tissue-specific transgene. Therefore, this system allows for tissueand cell-specific targeted infection of mammalian cells through ectopic expression of the viral receptor. Alternatively, when targeted infection of mammalian cells is not required (e.g., in cell culture), infection can be achieved through the use of non-avian envelopes, such as the amphotropic envelope from murine leukemia virus. The receptor for this envelope is endogenously expressed on almost all mammalian cells. We have used the RCASBP/RCANBP family of retroviral vectors extensively in both cultured cells and live animals for studies of viral replication and of cancer modeling in mice. Most of these studies have analyzed gain-of-function phenotypes by delivering and overexpressing a particular gene of interest. Recently, we engineered the RCANBP vector to reduce the expression of specific genes through the delivery of short hairpin RNA sequences. We also engineered this vector to control the expression of the inserted gene using the tetracycline (tet)- regulated system. Sequences inserted into this region are transcribed from a tet-responsive element and not the viral LTR. This virus allows inserted genes to be turned on and off in order to determine if expression of the gene is required for tumor initiation, maintenance, and progression. The ability to turn off gene expression will help determine if that gene is a good target for therapy. 31

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