Modulation of miR-146b expression by gene editing with CRISPR/Cas9 in papillary thyroid carcinoma

• Grant number: 24/04546-2
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: August 01, 2024
• End date: July 31, 2025
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Edna Teruko Kimura
• Grantee: Geovanna de Paula
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

The CRISPR/Cas9 methodology has emerged as a promising tool for the precise modification of DNA sequences throughout the genome. Recently, the Casgevy, an CRISPR/Cas gene editing therapy in patients with sickle cell anemia and thalassemia, was approved in the United Kingdom. In the oncological context, CRISPR/Cas9 has been applied in the experimental study of cancers including thyroid cancer (TC) in modulating coding and non-coding genes in in vitro and animal models. Papillary thyroid carcinoma (PTC) is the most frequent histotype and presents alterations in both coding and non-coding genes, such as microRNAs (miRNAs). Among all dysregulated miRNAs in PTC, miR-146b is the most highly expressed, being associated with tumor aggressiveness and survival. In this project, we aim to evaluate the silencing of the oncomiR miR-146b through gene editing using different sgRNAs to modify the MIR146B gene, located on chromosome 10, employing the CRISPR/Cas9 system in a PTC cell line. We will analyse the level expression level of genes modulated by miR-146b in different clones generated after silencing. In addition to assessing the effect of silencing on miR-146b expression, we will investigate the influence on cell proliferation, migration, and viability. The application of CRISPR/Cas9 to modulate miR-146b opens up a new possibilities for understanding the role of this microRNA in the tumor biology of PTC and its clinical applications in the future.