Evaluation of circulating transcript expression of the chaperone ERP29 in patients with head and neck cancer

• Grant number: 25/08498-5
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: August 01, 2025
• End date: July 31, 2026
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Gustavo Jacob Lourenço
• Grantee: Rodrigo Costa Cespedes
• Host Institution: Faculdade de Ciências Médicas (FCM). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil

Abstract

Chaperone proteins, such as the one encoded by the ERP29 gene, are crucial for protein folding and secretion from the endoplasmic reticulum to the Golgi complex. Failures in this process compromise protein functionality and cellular homeostasis. Studies conducted by our group indicated that the suppression of ERP29 in head and neck tumor cell lines is associated with increased expression of genes belonging to the MAPK and Akt pathways, which are known for their role in cell proliferation and inflammatory processes. However, the mechanisms involved in these processes remain uncertain. Thus, the objectives of this study are to evaluate the expression of circulating ERP29 transcripts in exosomes from patients with head and neck cancer (HNC) and to investigate the association of these transcripts with the patients' clinical and tumor characteristics, as well as their survival. For this purpose, 136 plasma samples from peripheral blood of HNC patients treated at the University Hospital of UNICAMP will be analyzed. Clinical data from the patients (age, sex, smoking, alcohol consumption, and inflammatory blood count data), tumor data (location, degree of differentiation, and TNM stage), and event-free survival (EFS) and overall survival (OS) information will be obtained from patient records. Exosomes will be isolated from patients' plasma using the Total Exosome Isolation Kit (from plasma) (Thermo Fisher Scientific). Exosomal RNA will be obtained using the Total Exosome RNA Isolation kit (Thermo Fisher Scientific) and will proceed to complementary DNA (cDNA) synthesis with the Maxima First Strand cDNA Synthesis Kit (Life Technologies), according to the manufacturer's recommendations. The determination of ERP29 transcript expression will be performed through qPCR using TaqMan assay reagents and the TaqMan Fast Advanced Master Mix reagent mixture (Thermo Fisher Scientific). The GAPDH gene will be used as the endogenous control in the qPCR reactions. Gene expression will be calculated using the arithmetic formula 2-¿¿Ct. If gene expression values follow a normal distribution, t-tests and ANOVA will be applied. Otherwise, Mann-Whitney and Wilcoxon tests will be used. EFS and OS will be estimated using the Kaplan-Meier method, and prognostic factors will be assessed through Cox regression analysis. The results of this study may contribute to identifying HNC patients who could benefit from personalized therapeutic approaches.