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Cancer Genes and Genomics

Hu/Mu ProtIn Oligonucleotide Microarray: Dual-Species Array for Profiling Protease and Protease Inhibitor Gene Expression in Tumors and Their Microenvironment

¹ Barbara Ann Karmanos Cancer Institute; Departments of ² Pharmacology, ³ Pathology, and ⁴ Obstetrics and Gynecology; ⁵ Applied Genomics Technology Core; and ⁶ Center for Molecular Medicine and Genetics and the Institute for Scientific Computing, Wayne State University, Detroit, Michigan; ⁷ Department of Cancer Biology, Vanderbilt University School of Medicine, Nashville, Tennessee; ⁸ Department of Pathology, Cancer Research Institute and Comprehensive Cancer Center, University of California, San Francisco, California; and ⁹ Proteases and Tissue Remodeling Unit, Oral and Pharyngeal Cancer Branch, National Institute of Dental and Craniofacial Research, NIH, Bethesda, Maryland

Requests for reprints: Donald R. Schwartz, Biodiscovery, LLC, 5692 Plymouth Road, Ann Harbor, MI 48105. Phone: 734-998-0751; Fax: 734-998-0750. E-mail: dschwartz{at}biodiscovery-llc.com

Proteolysis is a critical regulatory mechanism for a wide variety of physiologic and pathologic processes. To assist in the identification of proteases, their endogenous inhibitors, and proteins that interact with proteases or proteolytic pathways in biological tissues, a dual-species oligonucleotide microarray has been developed in conjunction with Affymetrix. The Hu/Mu ProtIn microarray contains 516 and 456 probe sets that survey human and mouse genes of interest (proteases, protease inhibitors, or interactors), respectively. To investigate the performance of the array, gene expression profiles were analyzed in pure mouse and human samples (reference RNA; normal and tumor cell lines/tissues) and orthotopically implanted xenografts of human A549 lung and MDA-MB-231 breast carcinomas. Relative gene expression and "present-call" P values were determined for each probe set using dChip and MAS5 software, respectively. Despite the high level of sequence identity of mouse and human protease/inhibitor orthologues and the theoretical potential for cross-hybridization of some of the probes, >95% of the "present calls" (P < 0.01) resulted from same-species hybridizations (e.g., human transcripts to human probe sets). To further assess the performance of the microarray, differential gene expression and false discovery rate analyses were carried out on human or mouse sample groups, and data processing methods to optimize performance of the mouse and human probe sets were identified. The Hu/Mu ProtIn microarray is a valuable discovery tool for the identification of components of human and murine proteolytic pathways in health and disease and has particular utility in the determination of cellular origins of proteases and protease inhibitors in xenograft models of human cancer. (Mol Cancer Res 2007;5(5):443–54)

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