ENRICHED LONG NON CODING RNAs SILENCING IN PANCREATIC CANCER STEM CELLS BY INDUCIBLE CRISPR-Cas13d SYSTEM

• Grant number: 23/08998-2
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: December 01, 2023
• End date: November 30, 2024
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Daniela Sanchez Basseres
• Grantee: Andressa Albuquerque Seiffert
• Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is an extremely lethal disease and accounts for more than 90% of pancreas tumors. The high lethality is due, in part, to the tumor intrinsic or acquired chemotherapeutic resistance and to the high metastatic potential of this type of cancer, with metastasis being an early event in the PDAC evolution and present in 50% of diagnosed patients. The tumor's high metastatic capacity and chemoresistance are associated with the acquisition and maintenance of a cancer stem phenotype by a population of cells known as cancer stem cells (CSC). CSC have epigenetic modifications that grant, among others, the tumor cells ability to migrate and invade, in addition to therapeutic resistance. Despite the importance of these cells for the biology of PDAC, the procedures involved in obtaining and maintaining the tumor stemness phenotype are still poorly understood. Based on the hypothesis that the epigenetic events involved in the acquisition of cancer stem cell phenotype may be mediated by long non-coding RNAs (lncRNAs), our group identified lncRNAs enriched in CSC. To study these RNAs functional roles in chemoresistance and metastasis, the silencing of these molecules will be performed by the CRISPR-Cas13d system, which combines the expression of a guide RNA complementary to the target of interest with the expression of Cas13d, a highly specific RNA endonuclease. Therefore, this project goal is to generate pancreatic tumor lines expressing Cas13d in a doxycycline inducible manner, which will subsequently be used to express the guide RNAs of interest. This model development and validation will be crucial for the functional role elucidation of the identified lncRNAs and, therefore, it is expected that this project will make an essential contribution to elucidate the mechanisms involved in PDAC malignancy, contributing to the development of more effective therapies.