Analysis of microRNome Expression (circulating and vesicular) and proteolytic signature in the plasma of patients with penile squamous cell carcinoma and prognostic evaluation.

• Grant number: 24/16137-0
• Support Opportunities: Regular Research Grants
• Start date: August 01, 2025
• End date: July 31, 2027
• Field of knowledge: Health Sciences - Medicine - Surgery
• Principal Investigator: Claudio Bovolenta Murta
• Grantee: Claudio Bovolenta Murta
• Host Institution: Instituto do Câncer do Estado de São Paulo Octavio Frias de Oliveira (ICESP). São Paulo , SP, Brazil
• Associated researchers: Alexis Germán Murillo Carrasco ; André Zelanis Palitot Pereira ; José Pontes Junior ; Kátia Ramos Moreira Leite ; Kayann Kaled Reda El Hayek ; Maria José Ferreira Alves ; Miyuki Uno ; Roger Chammas ; Tatiane Katsue Furuya ; William Carlos Nahas

Abstract

Analysis of microRNome Expression (circulating and vesicular) and proteolytic signature in the plasma of patients with penile squamous cell carcinoma and prognostic evaluation.INTRODUCTION: Penile cancer, although rare in Western countries, accounts for up to 20% of neoplasms affecting the male population in developing countries. The main prognostic indicators include pathological stage and tumor grade. While useful in clinical practice, these factors often perform below expectations. Surgical treatment for localized tumors involves penectomy, along with inguinal lymphadenectomy, resulting in high rates of recurrence-free survival. However, when lymph node involvement occurs, the results of surgical treatment are less satisfactory. To date, no effective systemic therapies exist, and response rates to radiotherapy are limited. The literature suggests that epigenetic phenomena, such as microRNA (miRNA) expression, play a significant role in the carcinogenesis of various neoplasms. Our research group has previously demonstrated that two miRNAs (miR-421 and miR-744-5p) are associated with metastasis in the tissues of patients with penile cancer. Following our initial studies, we hypothesize that these potential new biomarkers in penile cancer (miR-421 and miR-744-5p) are also overexpressed in the blood of patients with metastatic disease. We further hypothesize that another miRNA expression profile in blood and extracellular vesicles, as well as a proteolytic profile in the blood of these patients, may be associated with poor tumor prognosis. We believe that understanding these patterns may lead to the identification of new prognostic markers.OBJECTIVES: To confirm the overexpression in plasma of the two miRNAs (miR-421 and miR-744-5p) overexpressed in the tissues of patients with metastatic disease. To identify differentially expressed miRNAs in plasma and extracellular vesicles between patients with metastatic versus localized disease. To evaluate a proteolytic profile in the plasma of patients with metastatic disease compared to patients with localized disease. As a secondary objective, we will correlate the expression of miRNAs and proteolytic profiles with classical prognostic factors in penile cancer. Thus, we may suggest prognostic biomarkers or target therapy.METHODS: Whole blood was collected from 30 patients with penile squamous cell carcinoma (SCC) operated on at our institution between July 2015 and January 2018. These patients were followed prospectively with physical examination and imaging studies for metastasis assessment. After penectomy, the patients' disease was classified according to the risk of metastasis following the European Association of Urology classification. Low-risk patients were only followed up, while those with intermediate or high-risk disease, as well as those with clinical lymphadenopathy, underwent bilateral inguinal lymphadenectomy. For each plasma sample, three aliquots will be separated: miRNA extraction, purification of extracellular vesicles, miRNA extraction, and proteomic analysis resulting from proteolytic activity in the plasma. Using TaqMan" Array Cards, the circulating and vesicular miRNAs differentially expressed between patients with metastatic and localized disease will be identified; and, using the TAILS technique, a proteolytic profile between these groups of patients will be identified. The initially identified miRNAs, in addition to miR-421 and miR-744-5p overexpressed in the tissues of metastatic patients, will be validated by qRT-PCR. Subsequently, the findings of the miRNAs and proteolytic profile will be associated with tumor grade, pathological stage, perineural invasion, angiolymphatic invasion, tumor recurrence, and recurrence-free, cancer-specific, and overall survival.KEYWORDS: Penile neoplasms, microRNA, Prognosis, Biomarkers. (AU)