Larissa Silva dos Santos1, Rosyana de Fátima Vieira de Albuquerque1, Alessandra Silva e Silva1, Luana Quadros de Souza Leão2, Cláudia Patrícia Mendes de Araújo 1, Kamila Pereira de Araújo1, Giovana Melo Marques1, Anderson Ferreira Gonçalves3, Ejandre Lunieres Santiago 3, Mikele Praia de Oliveira3, Flávia Níniver de Oliveira Gomes3, Kátia Luz Torres3, Priscila Ferreira de Aquino1
Introduction: Cervical cancer is the most common type of cancer among women in Amazonas and the main cause is persistent infection by the human papillomavirus (HPV) with a high oncogenic risk, such as HPV-16 and HPV-18. This type of cancer is considered preventable, as it is preceded by precursor lesions (CIN2/3) which, when not detected and treated, can progress to cancer. Through excisional treatment, called conization, women should be monitored every 6 months for 1 or 2 years, based on the results of the cytopathological examination. It is estimated that 5% to 25% of these patients may have residual disease or recurrence of CIN 2/3, as well as having a five times greater risk of developing cancer. Therefore, the search for information that can help with diagnosis and prognosis is fundamental, and in this sense, the proteomic approach can provide relevant molecular information and help predict the risk of recurrence and progression. This study aimed to identify proteins with differential abundance that could be promising biomarkers in risk stratification for lesion recurrence or progression in a cohort of women. Material and Methods: To investigate these changes, cervical exfoliated cells from a cohort of 9 women were analyzed. Samples were collected using the Liqui-PREP® kit before and six months after treatment for CIN2/3. HPV 16/18 genotypes were detected by qPCR. For proteomic analysis, the samples were prepared using the bottom-up proteomics approach, where the peptides obtained were analyzed by LC-MS/MS on the Orbitrap Exploris 480 system coupled to liquid nanochromatography. The proteins were identified and quantified using PatternLab for Proteomics® software, and the Proteomic Pairwise Comparer was used for intra-individual comparative analysis. Results and discussion: The median age of the women in our cohort was 35 years. About cytopathology before treatment, all had a high-grade lesion and the histopathological diagnosis was 33% CIN 2, 56% CIN 3. HPV 16 was detected in 9 women (100%). After treatment, 9 (100%) women showed no cytomorphologic changes. The persistence of HPV 16 was observed in 2 (22%) after 6 months. Of these, 1 (11%) showed progression to early-stage cervical cancer (FIGO-IB1 staging) before 12 months after treatment. From the paired proteomic analysis, 167 proteins were identified and among them the ANXA4 and FBLN1 proteins showed significant changes (z-score> 2) in the patient who showed progression compared to the others, z-score of 8,92 and 10,16, respectively. Studies have shown that both ANXA4 (linked to cell proliferation and migration) and FBLN1 (involved in tissue remodeling) were increased in abundance in various epithelial tumors, including colorectal and breast cancer, and this increase may therefore reflect the activation of pathways associated with tumor progression. Conclusion: Thus, our study revealed relevant molecular changes after excisional treatment of CIN2/3, where proteins associated with tissue remodeling and cell proliferation were differentially abundant in the case of progression. These findings highlight the potential of proteomics as a predictive tool in post-treatment surveillance and early monitoring of recurrence.
Agradecimentos: PROEP/ILMD-FIOCRUZ AMAZÔNIA – LDMAIS; Amazonas State Oncology Control Center Foundation (FCECON-AM); Carlos Chagas Institute (ICC/Fiocruz PR); Institutional Scientific Initiation Scholarship Program (PIBIC)-CNPq; POSGRAD Program/FAPEAM;