27 - 30 May 2005

MOLECULAR PHENOTYPING OF MID-SECRETORY ENDOMETRIUM IN WOMEN WITH VERSUS WITHOUT SEVERE ENDOMETRIOSIS JLC Giudice [1], AE Hamilton [1], S Talbi [1], S Tulac [1], AP Hess [1], KC Vo [1], CN Nezhat [1], R Kempson [2], BA Lessey [3]. Department [1] OB/GYN and [2] Pathology Stanford University Stanford, CA USA [3] Reproductive Medicine Division Greenville Hospital System Greenville, SC USA

Endometriosis is a benign disorder associated with infertility and pelvic pain. The infertility is believed to be due to a variety of causes, including abnormal eutopic endometrium and an inhospitable environment for embryonic implantation.

We have previously reported on global gene profiling of human endometrium during the window of implantation in women with vs. without mild endometriosis (Kao et al, Endocrinology 2003;144:2870-2881). The results demonstrated numerous genes and gene families that were dysregulated in eutopic endometrium of women with disease compared to those without.

Most recently we have extended this study to eutopic endometrium of women with severe endometriosis, documented by laparoscopy. Cycle stage was confirmed by two independent pathologists, and the secretory phase was further subdivided, based on histologic evaluation. Endometrial biopsy samples were processed individually, poly (A)+ -RNA isolated, cDNA synthesized, and derived biotinylated cRNAs were then hybridized to the Affymetrix Human Genome U133A Plus 2.0 high-density oligo-nucleotide GeneChip arrays containing 54,000 probe sets, representing over 38,500 well characterized genes. The data then were extracted, the quality control assessed, and the data were subsequently exported to GeneSpring v6.2 (Silicon Genetics) for normalizations and further statistical analysis. A t-test statistic was applied to the data and further filtered based on flag presence, the control signal level, and a 1.5 fold change in gene expression. Nine hundred eleven probes showed differential expression between the samples from women with severe endometriosis vs. those without disease, with > 95% confidence and a fold change of 1.5 or more. Among the most up-regulated genes were fos and jun, gselect rowth factors and growth factor receptors, proteolytic enzymes, products of activated B-cells and T-cells, members of the Wnt signaling family, hedgehog receptor, calcium binding proteins, chemokine ligands and receptors, dual specificity phosphatase, PDE4B, gastrin, transcobalamin, perlecan, CDKN1C, prostaglandin E receptor 4, solute carrier proteins, and ER?.

Among the most highly down-regulated genes were: glutathione transferase GSTT1, matrix degrading and other proteolytic enzymes, heparinase, growth factors and cytokines, a variety of receptors (steroid hormone, growth factor, cytokine, protein hormone), extracellular matrix and cell surface glycoproteins, ion and water channels, select solute carriers, select transcription factors (p53), cell cycle regulators, signal transduction mediators, (e.g., forkhead family members), superoxide dismutase, inhibitor of coagulation, and immune modulators.

The data underscore a profound disturbance of endometrial gene expression in the window of implantation in women with vs. without disease, likely contributing to their infertility and to the pathogenesis of the disorder. Supported by NIH Specialized Cooperative Centers Program in Reproduction Research HD #31398 (LCG) and the NIH Office of Women’s Health Research (LCG).

List of abstracts from the 3rd International Conference on the Female Reproductive Tract