The role of p73 isoforms in the response of B16F10 cells to transduction with adenoviral vectors carrying p19Arf and IFNbeta cDNAs.

• Grant number: 18/04800-5
• Support Opportunities: Scholarships in Brazil - Post-Doctorate
• Effective date (Start): July 01, 2018
• Effective date (End): October 31, 2019
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Bryan Eric Strauss
• Grantee: Fernanda Antunes
• Host Institution: Instituto do Câncer do Estado de São Paulo Octavio Frias de Oliveira (ICESP). Coordenadoria de Serviços de Saúde (CSS). Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil
• Associated research grant: 15/26580-9 - Cancer gene therapy: strategic positioning for translational studies, AP.TEM

Abstract

Although metastatic melanoma is the least prevalent among skin cancers, it is the most lethal due to resistance to currently available treatments. Therefore, new effective therapeutic approaches are urgently needed. An interesting feature of melanoma is the presence of wild-type p53 protein, which in spite of being inactive represents an important therapeutic target since its modulation may promote reactivation of pro-death p53 signaling. Previous data from our group have shown that gene transfer of p19Arf, a functional partner of p53, and IFN², an inflammatory cytokine, acted synergistically in the induction of multimodal cell death, with characteristics of apoptosis and necroptosis, and also culminates in the release of immunogenic factors, promoting attraction of neutrophils, NK cells and CD4+ and CD8+ T lymphocytes. In addition, the modulation of 1054 genes was identified only in presence of the combination of p19Arf and IFN², and not by each one alone. Among these genes, TP73, which encodes p73 protein that belongs to the p53 and p63 protein family, stood out due to the remarkable increase in its expression. Since p73 has significant functional similarity to p53 and also acts in several cellular process such as cell death, autophagy, metabolism, among others, a better understanding of its signaling and also interaction with different cellular processes after p19Arf and IFN² treatment is necessary to elucidate possible molecular mechanisms elicited by this combination upon the treatment of melanoma. For this, the project presented here aims to use important and innovative tools, including B16F10 where p73 will be deleted using the CRISPR/Cas9 system, and also the development of a B16F10 cell line that is resistant to death caused by p19Arf and IFN² vectors. With this project, we aim to establish the involvement of p73 in the response of B16F10 cells upon delivery of the p19Arf and IFN² genes.