Modulation of miR-145-5p and miR-146b-5p levels is linked to reduced parasite load in H9C2 Trypanosoma cruzi infected cardiomyoblasts

Abstract

In the heart tissue of acutely Trypanosoma cruzi-infected mice miR-145-5p and miR-146b-5p are, respectively, downregulated and upregulated. Here, we used the H9C2 rat cardiomyoblast cell line infected with the Colombian T. cruzi strain to investigate the parasite-host cell interplay, focusing on the regulation of miR-145-5p and miR-146b-5p expression. Next, we explored the effects of interventions with the trypanosomicidal drug Benznidazole (Bz) alone or combined with Pentoxifylline (PTX), a methylxanthine derivative shown to modulate immunological and cardiac abnormalities in a model of chronic chagasic cardiomyopathy, on parasite load and expression of miR-145-5p and miR-146b-5p. The infection of H9C2 cells with trypomastigote forms allowed parasite cycle with intracellular forms multiplication and trypomastigote release. After 48 and 144 h of infection, upregulation of miR-145-5p (24 h: 2.38 ± 0.26; 48 h: 3.15 ± 0.9-fold change) and miR-146b-5b (24 h: 2.60 ± 0.46; 48 h: 2.97 ± 0.23-fold change) was detected. The peak of both miRNA levels paralleled with release of trypomastigote forms. Addition of 3 µM and 10 µM of Bz 48 h after infection reduced parasite load but did not interfere with miR-145-5p and miR-146b-5p levels. Addition of PTX did not interfere with Bz-induced parasite control efficacy. Conversely, combined Bz + PTX treatment decreased the levels of both microRNAs, resembling the expression levels detected in non-infected H9C2 cells. Moreover, the use of miR-145-5p and miR-146b-5p mimic/inhibitor systems before infection of H9C2 cells decreased parasite load, 72 h postinfection. When H9C2 cells were treated with miR-145-5p and miR-146b-5p mimic/inhibitor 48 h after infection, all the used systems, except the miR-146b-5p inhibitor, reduced parasite load. Altogether, our data indicate that these microRNAs putatively control signaling pathways crucial for parasite–host cell interaction. Thus, miR-145-5p and miR-146b-5p deserve to be further investigated as biomarkers of parasite control and tools to identify therapeutic adjuvants to etiological treatment in Chagas disease.

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

“…Lastly, we validated miR-146b-5p in individual samples, based on differential expression found in the acute phase of T. cruzi infection [38], crucially shown in in vitro model of T.cruzicardiomyocyte interaction [76]. miR-146b-5p was found upregulated in acutely infected mice [38] and here in the vehicle-treated infected group, supporting the consistent effect of infection on miR-146b-5p expression in both acute and chronic phases of CD.…”

“…The effects of Bz+PTX therapy on CCC were also previously explored by our group, supporting reduction in myocarditis and fibrosis, improvement of ECG abnormalities, reduction of TNF, and TNFR1 expression on CD8 + T cells and, more importantly, these effects were sustained after 30 days of treatment suspension [26]. Additionally, we recently assessed miR-146b-5p expression in T. cruzi-infected H9C2 cardiac cells, showing upregulation of this miRNA in early hours of infection with expression restoration upon Bz+PTX therapy, corroborating the participation of this miRNA in the parasite-host interaction process [76].…”

MicroRNA-145 enhances lung cancer cell progression after exposure to lyophilized fertile hydatid cyst fluid ofEchinococcus granulosussensu stricto

“…Hadighi et al, (2022) have shown that assessment of miR-145 in the plasma level of Iranian Plasmodium vivax malaria patients can introduce a promising diagnostic biomarker [20]. Farani et al, (2022) have also shown that over-expression of miR-145-5p and miR-146b-5p is related to decreased parasite burden in Trypanosoma cruzi -infected cardiomyoblasts which can candidate as promising biomarkers of parasite control to apply treatment methods in Chagas disease [21]. As well, Mohammadi et al, (2021) have shown that microRNA-365 enhanced apoptosis in A375 human melanoma cells line treated with fertile HCF [5].…”

“…Plasmodium vivax malaria patients can introduce a promising diagnostic biomarker [20]. Farani et al, (2022) have also shown that over-expression of miR-145-5p and miR-146b-5p is related to decreased parasite burden in Trypanosoma cruzi-infected cardiomyoblasts which can candidate as promising biomarkers of parasite control to apply treatment methods in Chagas disease [21].…”

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

“…When the mimic/inhibitor systems were applied 48 h after infection, all systems except the miR-146b-5p inhibitor resulted in a reduction in parasite load. These findings suggest that miR-145-5p and miR-146b-5p may play a role in controlling signaling pathways crucial for interacting with the parasite and host cells [ 23 ]. Therefore, further investigation of these miRNAs is warranted as potential biomarkers for parasite control and tools to identify therapeutic adjuvants for etiological treatment in ChD.…”