Abstract PO049: Compartment-specific profiling for evaluation of disrupted signaling in early stage, low-grade endometrial cancer

Abstract

Abstract Understanding tumor microenvironment is critical to improving conservative management (non-surgical) strategies for early stage, low grade endometrial cancer (EC). This remains an unmet clinical need for women who wish to maintain fertility and those who cannot undergo surgery due to medical co-morbidities. Current regimens rely on progestin treatment, but resistance and early relapse require improved therapeutic strategies. We sought to evaluate alterations in gene expression in the epithelial and stromal compartment of EC compared to normal endometrium to identify targets for combination therapies. RNA was extracted from microdissected epithelial and stromal compartments of formalin fixed paraffin embedded early stage, low grade EC and normal endometrium (NE). Comprehensive transcriptome profiling was performed with Affymetrix Clariom D assay and pathway analysis was performed. Immunohistochemistry (IHC) was used for validation. Functional studies of differentially expressed genes were performed in Hec1A EC cell lines. Among differentially expressed genes in EC vs NE, multiple altered signaling pathways were identified. EIF2 signaling, eIF4 and p70S6K signaling, and PI3K/AKT/mTOR were the most significant pathways activated in both epithelial and stromal compartments. In the epithelia compartment, the most significant gene expression changes included reduced expression of RBPMS (RNA Binding Protein with Multiple Splicing), SORD (Sorbitol Dehydrogenase), DDX17 (DEAD-Box Helicase 17), and TPT1 (Translationally-Controlled Tumor Protein) in EC, while MTDH (Metadherin) was significantly increased in EC. Similar changes were observed in the stroma compartment, but with lower fold changes than that observed in gland. Together with decreased RBPMS expression, decreased RBPMS-interacting molecules IGF2, TCF7L2 (Transcription Factor 7 Like 2), FOS (Proto-Oncogene C-Fos) and increased GPS2 (G protein Pathway Suppressor 2) were also observed. Differential expression of proteins in gland and stroma was also confirmed by IHC. An inverse correlation of RBPMS expression and EC proliferation was also observed in EC at the protein level. Enhanced AKT and S6 protein phosphorylation were observed by RBPMS silencing in Hec1A endometrial cells. Reverse Phase Protein Array demonstrated activation of Caveolin 1, FOXO3 (Forkhead Box O3), MT-CO1 (Mitochondrially Encoded Cytochrome C Oxidase I), L1CAM (L1 Cell Adhesion Molecule) with RBPMS silencing in Hec1A cell. These molecules are involved in pathways such as molecular mechanisms of cancer, estrogen receptor signaling, and senescence pathway. While estrogen-regulated changes were prominent in early stage low-grade EC, the identification of additional signaling changes provide support for an ongoing trial of levonorgestrel IUD + everolimus (mTOR inhibitor) and can be probed further to identify new targets for combination conservative management strategies. Additionally, biomarkers from these pathways, such as RBPMS, can be studied to guide the selection of progestin-only therapy versus combination therapy. Citation Format: Qian Zhang, Xiaoping Su, Joseph Celestino, Ying Yuan, Samuel C. Mok, Karen H. Lu, Melinda S. Yates. Compartment-specific profiling for evaluation of disrupted signaling in early stage, low-grade endometrial cancer [abstract]. In: Proceedings of the AACR Virtual Special Conference: Endometrial Cancer: New Biology Driving Research and Treatment; 2020 Nov 9-10. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(3_Suppl):Abstract nr PO049.