Treatment with benznidazole and pentoxifylline regulates microRNA transcriptomic profile in a murine model of Chagas chronic cardiomyopathy

Abstract

Chronic Chagas cardiomyopathy (CCC) is one of the leading causes of morbidity and mortality due to cardiovascular disorders in endemic areas of Chagas disease (CD), a neglected tropical illness caused by the protozoan parasite Trypanosoma cruzi. CCC is characterized by parasite persistence and inflammatory response in the heart tissue, which occur parallel to microRNA (miRNA) alterations. Here, we investigated the miRNA transcriptome profiling in the cardiac tissue of chronically T. cruzi-infected mice treated with a suboptimal dose of benznidazole (Bz), the immunomodulator pentoxifylline alone (PTX), or the combination of both (Bz+PTX), following the CCC onset. At 150 days post-infection, Bz, PTX, and Bz+PTX treatment regimens improved electrocardiographic alterations, reducing the percentage of mice afflicted by sinus arrhythmia and second-degree atrioventricular block (AVB2) when compared with the vehicle-treated animals. miRNA Transcriptome profiling revealed considerable changes in the differential expression of miRNAs in the Bz and Bz+PTX treatment groups compared with the control (infected, vehicle-treated) group. The latter showed pathways related to organismal abnormalities, cellular development, skeletal muscle development, cardiac enlargement, and fibrosis, likely associated with CCC. Bz-Treated mice exhibited 68 differentially expressed miRNAs related to signaling pathways like cell cycle, cell death and survival, tissue morphology, and connective tissue function. Finally, the Bz+PTX-treated group revealed 58 differentially expressed miRNAs associated with key signaling pathways related to cellular growth and proliferation, tissue development, cardiac fibrosis, damage, and necrosis/cell death. The T. cruzi-induced upregulation of miR-146b-5p, previously shown in acutely infected mice and in vitro T. cruzi-infected cardiomyocytes, was reversed upon Bz and Bz+PTX treatment regimens when further experimentally validated. Our results further our understanding of molecular pathways related to CCC progression and evaluation of treatment response. Moreover, the differentially expressed miRNAs may serve as drug targets, associated molecular therapy, or biomarkers of treatment outcomes.

Unraveling the role of miRNAs as biomarkers in Chagas cardiomyopathy: Insights into molecular pathophysiology

“…Considering the assessments of expression level, a total of 2,311 miRNAs were evaluated ( Table 1 ). Silva Grijó Farani and colleagues [ 25 ] and Laugier and colleagues [ 20 ] utilized comparable commercial miRNA array panels comprising 752 miRNAs from the same manufacturer. Conversely, Nonaka and colleagues (2021) [ 21 ] investigated 88 miRNAs, while Navarro and colleagues [ 22 ] examined 642 miRNAs using quantitative real-time reverse-transcription PCR (qRT-PCR) arrays.…”

“…A recent study conducted by Silva Grijó Farani and colleagues [ 25 ] also investigated the expression profiles of 752 miRNAs by qRT-PCR array in the cardiac tissue of mice with chronic T . cruzi infection treated with Bz, PTX, or a combination of both (Bz + PTX).…”

“…Since miR-21 is associated with fibrosis and immune response, targeting miR-21 could be a promising therapeutic for ChCM [ 12 , 13 , 21 , 22 ]. The miR-146b was up-regulated during the acute phase of ChD specifically in cardiac tissue, and it seems to play a regulatory role in the interaction between the parasite and the host cell, suggesting its potential as a biomarker for therapeutic response [ 22 , 23 , 25 ]. Furthermore, both miR-155 and miR-146a are consistently up-regulated in plasma and cardiac tissue samples during the acute and chronic phases of ChD.…”

“…A recent study conducted by Silva Grijo ´Farani and colleagues [25] also investigated the expression profiles of 752 miRNAs by qRT-PCR array in the cardiac tissue of mice with chronic T. cruzi infection treated with Bz, PTX, or a combination of both (Bz + PTX). After 150 days of infection, the Bz + PTX-treated group showed 58 DEMs associated with key signaling pathways related to cellular growth, tissue development, cardiac fibrosis, damage, and cell death.…”

Quantitative PCR as a marker for preemptive therapy and its role in therapeutic control in Trypanosoma cruzi/HIV coinfection

“…Therefore, the search for early markers of the evolution of clinical forms represents a PLOS NEGLECTED TROPICAL DISEASES challenge. MicroRNAs have been highlighted but not tested yet in human disease or immunocompromised individuals [51][52][53].…”