EFEITOS CLÍNICOS, BIOQUÍMICOS E MOLECULARES EM PORTADORES DE FIBRODISPLASIA OSSIFICANTE PROGRESSIVA ANTES E DEPOIS DO TRATAMENTO COM ÁCIDO ASCÓRBICO, PROPRANOLOL E HESPERIDINA

Abstract

Fibrodysplasia Ossificans Progressiva (FOP) is a congenital, genetic, and rare disease. It is characterized by irreversible and untreatable skeletal malformations, with devastating heterotopic ossifications, and currently, there is no cure. Anti-inflammatory therapies show potential for improvement and control of acute crises. Treatment with ascorbic acid combined with propranolol (FOPCON) has demonstrated benefits for FOP symptoms due to its antioxidant, anti-inflammatory, and anti-angiogenic properties. Hesperidin is an antioxidant flavonoid with anti-inflammatory effects; however, its mechanism of action, when combined with ascorbic acid in the stabilization of FOP in some patients, is unknown, considering that access to tissue studies from FOP patients is practically unfeasible. Objective: to analyze the mRNA expression of four target genes involved in the pathophysiology of FOP, namely COL1, ADRB2, TNFα, and RUNX2, in buccal epithelial cells (BEC) of patients treated with FOPCON (n = 8), patients treated with FOPCON+H (n = 7), versus healthy controls (n = 5), following treatment for six months. The results of this study were compared with samples of peripheral blood mononuclear cells (PBMC) from previous studies. Method: Total RNA was extracted, followed by reverse transcription and real-time PCR. The relative expression profile of four genes in buccal epithelial cells from FOP patients versus controls was significantly different. An unpaired t-test and ANOVA were used. Results: Basal mRNA expressions confirmed significant differences in the expression profiles of TNF-α (p < 0.05) and COL1 (p < 0.05) genes and also for the ADBR2 (p < 0.05) and RUNX2 (p < 0.05) genes, which previously only showed an increasing trend. The treatments of FOP patients with the FOPCON compound (AA+PP) and FOPCON Hesperidin (AA+PP+H) showed effects on the gene expression of COL1, TNF-α, ADBR2, and RUNX2 in FOP, characterizing a distinct phenotypic profile of FOP BEC. Conclusion: The treatment had an effect on gene modulation. FOPCON (ascorbic acid + propranolol) allowed gene modulation even with the addition of Hesperidin. Although there was no sensitivity in the method capable of differentiating the enhancing effect of Hesperidin, there was no harm in the treatment with FOPCON, and it maintained the same condition in terms of gene expression. The use of buccal epithelial cells becomes a potential material, both for evaluating treatment in its clinical aspect and for obtaining important molecular data related to the pathophysiolo of FOP.

Keywords: Fibrodysplasia Ossificans Progressiva; FOP mRNA expression; epithelial cells; buccal mucosa; squamous cell scrapings.