Reprogramming Macrophages through CRISPR-Cas9: an alternative to treating tumors

• Grant number: 23/15440-8
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: November 01, 2024
• End date: October 31, 2027
• Field of knowledge: Biological Sciences - Immunology - Cellular Immunology
• Principal Investigator: Rodrigo Nalio Ramos
• Grantee: Carla Sanzochi Fogolin
• Host Institution: Hospital das Clínicas da Faculdade de Medicina da USP (HCFMUSP). Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil

Abstract

Macrophages play diverse roles in the body, including modulation of tissue homeostasis, antigen presentation, phagocytosis, and pathogen elimination. There are two main macrophage differentiation pathways: embryonic-derived macrophages, which become tissue-resident, and blood-derived macrophages recruited during inflammatory processes. In the tumor microenvironment, macrophages are one of the main components of the immune infiltrate and conventionally play immunosuppressive and pro-tumorigenic roles. High cellular density of tumor-associated macrophages (TAMs) in various tumors correlates with poor prognosis. Additionally, TAMs can promote tumor growth, angiogenesis, and metastasis. The heterogeneity of these cells and their complexity have been explored with technologies such as Single Cell RNA-seq, which revealed a broad spectrum of macrophage subtypes, associated with different spatial distributions in tissues and varied functions. Currently, it is possible to genetically manipulate macrophages in vitro with the CRISPR-Cas9 gene editing approach, modulating target genes. Thus, we believe that modulating genes associated with macrophage functions could modify the functional states of these cells and provide a major gain in tumor control. Therefore, using bioinformatics tools, we aim to 1) identify and select genes expressed in macrophages and 2) induce their silencing by CRISPR-Cas9. The main objective of the project is to induce the reprogramming of macrophages with phenotypes similar to TAMs by means of CRISPR-Cas9 targeting new targets derived from an integrative analysis of transcriptomic and proteomic data. With the implementation of gain versus loss of function mechanistic assays, we aim to understand the molecular events of macrophage reprogramming and its application in new antitumor therapies.