In vitro study of the Sonic Hedgehog signaling pathway inhibitors in uterine leiomyoma and leiomyosarcoma

• Grant number: 15/23482-6
• Support type: Scholarships in Brazil - Doctorate
• Effective date (Start): March 01, 2016
• Effective date (End): October 31, 2019
• Field of knowledge: Health Sciences - Medicine - Maternal and Child Health
• Principal researcher: Kátia Cândido Carvalho
• Grantee: Natália Garcia
• Home Institution: Faculdade de Medicina (FM). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated scholarship(s): 17/24448-1 - In vitro and in vivo evaluation of SHH inhibitors effects on uterine leiomyoma and leiomyosarcoma, BE.EP.DR

Abstract

The leiomyoma and leiomyosarcoma are tumors of mesenchymal origin that develop in the uterus. These tumors have variable clinical behavior, compromising fertility and may even lead to death. The leiomyoma is a benign tumor commonly found in women of reproductive age. Leiomyosarcoma represents about 40% of sarcomas of the uterus. leiomyosarcoma and leiomyomas 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 Hedgehog signaling pathway Sonic (SHH) is related to the development of different types of cancer, since it plays important role in cell proliferation and differentiation. In a previous study, our group evaluated the expression profile of molecules involved in the SHH pathway in mesenchymal tumors of the uterus. The SMO, GLI1 and SHH proteins had over expression in leiomyosarcomas, down expression in leiomyomas and absent in adjacent normal myometrium. These results aroused our interest in evaluating the therapeutic potential inhibitors of the pathway in these tumors, because some of these proteins have been inhibited in other cancers with very promising results. Thus, this project aims to evaluate in vitro the action of signaling pathway inhibitors of the Sonic Hedgehog in leiomyoma and leiomyosarcoma. To do this, initially, cell lines will be used which will be submitted to specific inactivation of the SHH pathway genes by RNAi or by treatment with specific inhibitors SMO (GDC 0449, LDE 225 and cyclopamine); GLI 1 (56 GANT, GANT 61, IPH IPH 1 and 6) and SHH (Robotinikinin). The cells will be after confirmation of inhibition of targets, assessed for capacity migration and invasion. From the data generated in the previous step of inhibition by specific drugs, the most effective inhibitors will be tested in primary culture of cells from up to 10 patients with leiomyoma and leiomyosarcoma, undergoing surgery fibroids Clinic of Disciplina de Ginecologia da Faculdade de Medicina da USP. The effectiveness of inhibitors in the cells will be evaluated by Western Blott and real time PCR for both the pathway molecules (SHH, PTCH1, SMO, GLI and Sufu 1-3) and for their targets (BMP4, BCL-2 and CCND1). All results will be analyzed statistically.