Effects of Epstein-Barr virus RPMS1 protein expression on lymphoid human cells cultured in vitro

• Grant number: 19/05061-4
• Support Opportunities: Scholarships abroad - Research Internship - Master's degree
• Start date: July 08, 2019
• End date: January 07, 2020
• Field of knowledge: Health Sciences - Medicine - Pathological Anatomy and Clinical Pathology
• Principal Investigator: Deilson Elgui de Oliveira
• Grantee: Beatrice Adrianne Silva Jorge
• Supervisor: Ethel Cesarman
• Host Institution: Instituto de Biotecnologia (IBTEC). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
• Institution abroad: Weill Cornell Medical College, United States
• Associated to the scholarship: 18/12164-1 - Effects of Epstein-Barr protein RPMS1 expression in human cells cultured in vitro, BP.MS

Abstract

The Epstein-Barr virus (EBV) - formally human gammaherpesvirus 4 (HHV-4) - is a herpes-virus associated with the development of a variety of human malignancies, notably Burkitt lym-phoma (BL) and nasopharyngeal carcinoma (NPC), but also subsets of cases for Hodgkin and non-Hodgkin lymphomas (particularly in immunocompromised patients) and gastric carcino-mas. World Health Organization data indicates that the highest incidence of BL occurs in re-gions of Central Africa and New Guinea, reaching 6,700 new cases annually. Almost all BL cases occurring on African continent are associated with EBV. Other types of lymphomas such as Hodgkin and Diffuse Large B cell represents a range between 30%-50% and 10% of the cases, respectively. The oncogenic potential of EBV is associated with viral products that interfere with critical biological processes, such as genetic stability, apoptosis regulation, and immortalization capacity. So far, the main EBV oncogenic product is the latent protein membrane 1 (LMP1), which causes constitutive activation of NF-kB, among other activities. Nonetheless, other viral products only recently have been investigated, such as the EBV RPMS1 protein, which is gener-ated by alternative splicing of the BamHI A right transcripts (BART) segment of the viral ge-nome. RPMS1 was reported to modulate the Notch pathway, and it may possibly contribute for the EBV-induced transformation of epithelial cells. For NPC, a A/G single nucleotide polymor-phism (SNP) at position 155391 in the RPMS1 coding sequence was associated with increased risk for the disease. The eventual role of RPMS1 in pathogenesis of EBV-associated lymphopro-liferative disorders is largely unknown. Thus, the present study aims to investigate the effects of RPMS1 knockdown on apoptosis, cell viability and protein expression in EBV-positive lymphoid cell lines. For that purpose, the expression levels of RPMS1 mRNA will be evaluated in EBV-positive lymphoid cell lines by real-time quantitative (RT-qPCR) and the cell lines that express the highest amount of RPMS1 mRNA will be selected for further assays. RPMS1 knockdown will be conducted by small interference RNA (siRNA) and assayed in vitro for cell viability and apop-tosis. Lastly, the expression of key NFkB pathway proteins will be analyzed by Western blot. As the outcome of this study, we expect to identify potential roles of RPMS1 on the oncogenic properties of EBV-positive lymphoid cell lines. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

CAETANO, BRUNNO FELIPE R.; JORGE, BEATRICE ADRIANNE S.; MUELLER-COAN, BARBARA GRASIELE; DE OLIVEIRA, DEILSON ELGUI. Epstein-Barr virus microRNAs in the pathogenesis of human cancers. Cancer Letters, v. 499, p. 14-23, FEB 28 2021. (19/05061-4, 17/20352-0, 17/22312-5, 19/03804-0, 18/12164-1)