Identification of HOXB2 gene targets in glioblastoma cell line by promotor capture Hi-C technique

• Grant number: 19/17348-6
• Support Opportunities: Scholarships abroad - Research Internship - Master's degree
• Effective date (Start): January 06, 2020
• Effective date (End): May 05, 2020
• Field of knowledge: Biological Sciences - Genetics - Human and Medical Genetics
• Principal Investigator: Wilson Araújo da Silva Junior
• Grantee: Natália Volgarine Scaraboto Bonfa
• Supervisor: Peter Fraser
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Research place: Florida State University, United States
• Associated to the scholarship: 17/24027-6 - Analysis of gene pathways regulated by the HOXB2 gene in glioblastoma, BP.MS

Abstract

Glioblastoma (GBM) is the most common and lethal neoplasm that affects astrocytic cells, with extremely aggressive and invasive growth. It is marked by low survival rates, usually just over a year. In this tumor, there is an intense deregulation in gene expression, highlighting the up regulation of 33 HOX genes. Of these, we found that high levels of expression of the HOXB2 gene are correlated with low GBM survival. Data from studies conducted as part of my master project reveals that the silencing of HOXB2 promotes drastic phenotypic alterations in two GBM cell lines, such as decrease in cell proliferation and cell cycle arrest. In this context, our main goal is to elucidate the targets of HOXB2, to better understand it role in gene regulation and GBM development. For this, we have applying ChiP-seq and RNA-seq techniques, however, they allow only partial evaluation of HOXB2 targets, and they do not distinguish direct and indirect targets. Therefore, we intend to perform Promotor Capture Hi-C (PCHi-C), a method developed from Hi-C that allows the genome-wide detection of distal promoter-interacting regions, through a capture system based on biotinylated RNA baits. Allied to the data from our ChiP-seq, the PCHi-C will provide a full elucidation of HOXB2 targets in GBM, with potential therapeutic insights. PCHi-C technique and analysis will be accomplished in collaboration with Dr. Peter Fraser, who worked on its development. (AU)