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2002 Scientific Report

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Analytical, Cellular,

Analytical, Cellular, and Molecular Microscopy Laboratory James H. Resau, Ph.D. Dr. Resau received his Ph.D. from the University of Maryland School of Medicine in 1985. Between 1968 and 1994, he was in the U.S. Army (active duty and reserve assignments) and served in Vietnam. From 1985 until 1992, Dr. Resau was a faculty member of the University of Maryland, School of Medicine, Department of Pathology, and was a tenured Associate Professor from 1990–1992. Dr. Resau then went to the NCI to be Director of the Analytical, Cellular and Molecular Microscopy Laboratory in the Advanced BioScience Laboratories–Basic Research Program at the National Cancer Institute–Frederick Cancer Research and Development Center, Maryland (1992–1999). Dr. Resau joined VARI as a Special Program Investigator in June 1999. Staff Bree Buckner, B.S., HTL (ASCP), QIHC Eric Hudson, B.S. J.C. Goolsby Laboratory Members Students Hien Dang Marie Graves Lateefah Gray Maketta Hassen Brandon Leeser Matthew Main Christine Moore Jeanine Myles Research Projects Our laboratory works closely with VARI investigators, as well as in collaboration with outside parties, to provide a number of microscopy needs. We have a special interest in the quantification of imagery. We have two confocal microscopes that enable us to visualize organelles and processes in cells and tissues such as receptor–ligand interactions and co-localization of proteins with organelles. We have studied the location of two gene-targeted proteins within a cell (i.e., GFP and RFP) with a DNA marker, DAPI, in three dimensions. We have integrated laser-capture microdissection instrumentation into the program, as well as paraffin and frozensection staining. We also provide histotechnology services, consultation on staining, and direction for the human tissue services. In collaboration with George Vande Woude and Rick Hay of VARI, we have developed the Tissue Collection Initiative between our Institute and the Spectrum Health System in Grand Rapids. We have expanded the program to include the Holland, Pennock, and Hackley hospitals. This program provides for the collection and characterization of fresh-frozen surgical tissues that will allow investigators to create a working repository for a wide range of projects. Surgically removed human tumors and normal tissue will be evaluated in institutional review board (IRB)–approved basic and translational research projects. This collection will at the same time provide to the physician researcher access to research collaborations with the intention of facilitating translation of new diagnostic, treatment, and evaluation protocols. We plan to generate gene expression profiles (microarray), establish new tumor cell lines, and develop new diagnostic and therapeutic agents through this collaboration. Epidemiologic evaluations will also be greatly improved by the coordination of clinical information, diagnosis, and research results. The goal of this project is to develop genetic-based diagnostic classification of human disease. There is a Scientific Advisory Board for this project comprising members of VARI and the Spectrum Health pathology, surgical and medical oncology, and surgery departments. Tissue is collected with explicit written permission of the participating physicians and patients. Protocols for the use of the material in this archive require the approval of both the VARI and the Spectrum Health IRBs. Directly related to this archive is the paraffinblock repository called SPIN (Shared Pathology Tissue and Informatics Network), a project that involves the same hospitals in west Michigan. This archive stores and catalogs paraffin blocks that are older than five years and ordinarily would be destroyed. Currently the archive holds approximately 150,000 tissue samples/paraffin blocks. They are not directly linked to any personal identifiers or names and there is limited demographic information available. Of the 150,000 samples or blocks, clinical and demographic data is available 37

for about 20%. The material from future years will be available with digital information on age, sex, and diagnosis. The current samples were collected by the hospitals with nondigital database technology, and the information is slowly being transcribed. These samples will be used in cellular and molecular protocols approved by our IRB. The samples and demographics are identified with basic information (such as diagnosis, age, sex, etc.) in a webbased, interactive format for determination of prognosis, diagnosis, and therapy. In the first six months of operation of SPIN there have been 12 users registered who have submitted 83 requests for searches and 33 subsequent tissue requests. We have provided tissue and histopathology services for 12 VARI investigators and have generated over 72,000 microscopic images and related image files for the collaborations. A major part of both the tissue initiative and SPIN programs will be to annotate and update information on each specimen. Our own research interest is in the retrospective review of archival tissues from individual samples with known clinical outcomes. We identify and quantify the location of particular proteins and examine the relationship between their pattern of expression and the prognosis of disease progression. We have submitted the results from our U.S. Army Breast Cancer–funded analysis of Met and Her2neu in human breast cancer and have nearly completed a major study in collaboration with investigators in Chicago on the same problem. The advantage of archival material is that new markers of prognosis can be evaluated. We can measure up to four proteins simultaneously, and we are developing methods to combine confocal microscopy, gene expression, and laser-capture microdissection with traditional surgical pathology diagnostic procedures to determine prognosis via an objective and quantifiable method(s). These tools—and, more importantly, the process—will have application to many types of human disease. We have recently obtained NIH funding for a major effort in multiphoton imaging of developmental and carcinogenic events in GFP-expressing transgenic mice. George Vande Woude, Ilan Tsarfaty, and I will evaluate the role of Met and HGF/SF in branching morphogenesis, carcinogenesis, and therapy of cancer-related compounds. Other collaborations within VARI involve Met and HGF-SF in cells and tissues; the role of Met and HGF-SF in prostate and breast carcinogenesis; the location of gene-targeted proteins in rodents; evaluation of monoclonal antibodies as diagnostic reagents; application of human tissues to molecular-based studies; and the cellular and subcellular localization and quantification of proteins. We also have ongoing collaborations with scientists in other countries. We have in development a molecular imaging project with scientists at Tel Aviv University in Israel, and we have an international breast cancer project to evaluate Her2/neu and Met/HGF interactions with scientists in Germany. In addition to our research program, we have a long-standing interest in science education. Together with Grand Valley State University and Grand Rapids Community College, we have received NIH funding as part of the Bridges to the Baccalaureate program to support the recruitment and graduation of women and minorities into science, mathematics, and research careers. Dr. Resau is a co-investigator and site coordinator for the Bridges program. External Collaborators Stephan Baldus, University of Cologne, Germany Maria Birchenall-Roberts, Francis Ruscetti, Jerrold Ward, and George Pavlakis, National Cancer Institute, Frederick, Maryland Ruth Heilmann, University of Chicago, Illinois Iafa Keydar and Ilan Tsarfaty, Tel Aviv University, Israel Justin McCormick, Michigan State University, Lansing Toshio Mura, National Cancer Institute, Bethesda, Maryland John Sacci, University of Maryland, Baltimore Duane Smoot, Howard University, Washington, D.C. 38

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