In vitro and in vivo evaluation of SHH inhibitors effects on uterine leiomyoma and leiomyosarcoma

Abstract

Leiomyoma (LM) and leiomyosarcoma (LMS) are mesenchymal tumors of uterine origin. LM and LMS are myometrial neoplasms that have the same pattern of cell differentiation, but with completely different clinical progression. Several studies have shown that aberrant activation of the Sonic Hedgehog signaling pathway (SHH) is related to the development of different types of cancer. In a previous study, our group evaluated the expression profile of the SHH pathway molecules in uterine LM and LMS. SMO and GLI1 proteins were overexpressed in LMS, down expressed in LM and absent in myometrium. These results aroused our interest in the therapeutic potential of this pathway inhibitors in these tumors, because some of these proteins have been inhibited in other cancers with very promising results. Thus, this project aims to evaluate the effects in vitro and in vivo of the SHH signaling pathway inhibitors in LM and LMS. To do this, initially, cell lines will be used for specific SMO and GLI1 inactivation (using iRNA/CRISPR-CAS9 or specific chemical inhibitors) or their overexpression. The inhibitory effects of the gene targets will be assessed by analysis of cell migration, proliferation and invasion. Animal tumor induction, for SMO and GLI1 block treatment, will be performed in Balb/c nude mouse. The mice will be randomized in groups and will receive treatment with specific drug or vehicle only. Tumor size development will be measured. We will analyze whether overexpression induction of SMO or GLI may lead to a possible tumor transformation in uterine smooth muscle cells in mice model. After treatment, the tumors will be removed and gene and protein expression will be evaluated for molecules of the Sonic Hedgehog pathway. All results will be analyzed statistically. (AU)