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Trypanosoma cruzi Dysregulates piRNAs Computationally Predicted to Target IL-6 Signaling Molecules During Early Infection of Primary Human Cardiac Fibroblasts

  • Ayorinde Cooley (Department of Microbiology, Immunology and Physiology, Meharry Medical College) ;
  • Kayla J. Rayford (Department of Microbiology, Immunology and Physiology, Meharry Medical College) ;
  • Ashutosh Arun (Department of Microbiology, Immunology and Physiology, Meharry Medical College) ;
  • Fernando Villalta (Department of Microbiology, Immunology and Physiology, Meharry Medical College) ;
  • Maria F. Lima (Department of Cell, Molecular, and Biomedical Sciences, School of Medicine, The City College of New York) ;
  • Siddharth Pratap (School of Graduate Studies and Research, Meharry Medical College) ;
  • Pius N. Nde (Department of Microbiology, Immunology and Physiology, Meharry Medical College)
  • Received : 2022.03.17
  • Accepted : 2022.10.26
  • Published : 2022.12.31
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Abstract

Trypanosoma cruzi, the etiological agent of Chagas disease, is an intracellular protozoan parasite, which is now present in most industrialized countries. About 40% of T. cruzi infected individuals will develop severe, incurable cardiovascular, gastrointestinal, or neurological disorders. The molecular mechanisms by which T. cruzi induces cardiopathogenesis remain to be determined. Previous studies showed that increased IL-6 expression in T. cruzi patients was associated with disease severity. IL-6 signaling was suggested to induce pro-inflammatory and pro-fibrotic responses, however, the role of this pathway during early infection remains to be elucidated. We reported that T. cruzi can dysregulate the expression of host PIWI-interacting RNAs (piRNAs) during early infection. Here, we aim to evaluate the dysregulation of IL-6 signaling and the piRNAs computationally predicted to target IL-6 molecules during early T. cruzi infection of primary human cardiac fibroblasts (PHCF). Using in silico analysis, we predict that piR_004506, piR_001356, and piR_017716 target IL6 and SOCS3 genes, respectively. We validated the piRNAs and target gene expression in T. cruzi challenged PHCF. Secreted IL-6, soluble gp-130, and sIL-6R in condition media were measured using a cytokine array and western blot analysis was used to measure pathway activation. We created a network of piRNAs, target genes, and genes within one degree of biological interaction. Our analysis revealed an inverse relationship between piRNA expression and the target transcripts during early infection, denoting the IL-6 pathway targeting piRNAs can be developed as potential therapeutics to mitigate T. cruzi cardiomyopathies.

Keywords

Acknowledgement

We are grateful to the Molecular Biology Core Facility and the Morphology Core at Meharry Medical College. This work was supported by NIH grants 1SC1AI27352 (PNN), 2T32AI007281-31 (AC), 2T32HL007737-26 (AC), 5R25GM05994 (KJR), F31AI167579 (KJR), and U54MD007586 (FV). The funders had no role in the study design, data collection, and analysis, decision to publish, or preparation of the manuscript.

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