Thrombospondin-1 Plays an Essential Role in Yes-Associated Protein Nuclear Translocation during the Early Phase of Trypanosoma cruzi Infection in Heart Endothelial Cells

Abstract

The protozoan parasite Trypanosoma cruzi is the causative agent of Chagas disease. This neglected tropical disease causes severe morbidity and mortality in endemic regions. About 30% of T. cruzi infected individuals will present with cardiac complications. Invasive trypomastigotes released from infected cells can be carried in the vascular endothelial system to infect neighboring and distant cells. During the process of cellular infection, the parasite induces host cells, to increase the levels of host thrombospondin-1 (TSP-1), to facilitate the process of infection. TSP-1 plays important roles in the functioning of vascular cells, including vascular endothelial cells with important implications in cardiovascular health. Many signal transduction pathways, including the yes-associated protein 1 (YAP)/transcriptional coactivator, with PDZ-binding motif (TAZ) signaling, which are upstream of TSP-1, have been linked to the pathophysiology of heart damage. The molecular mechanisms by which T. cruzi signals, and eventually infects, heart endothelial cells remain unknown. To evaluate the importance of TSP-1 expression in heart endothelial cells during the process of T. cruzi infection, we exposed heart endothelial cells prepared from Wild Type and TSP-1 Knockout mouse to invasive T. cruzi trypomastigotes at multiple time points, and evaluated changes in the hippo signaling cascade using immunoblotting and immunofluorescence assays. We found that the parasite turned off the hippo signaling pathway in TSP-1KO heart endothelial cells. The levels of SAV1 and MOB1A increased to a maximum of 2.70 ± 0.23 and 5.74 ± 1.45-fold at 3 and 6 h, respectively, in TSP-1KO mouse heart endothelial cells (MHEC), compared to WT MHEC, following a parasite challenge. This was accompanied by a significant continuous increase in the nuclear translocation of downstream effector molecule YAP, to a maximum mean nuclear fluorescence intensity of 10.14 ± 0.40 at 6 h, compared to wild type cells. Furthermore, we found that increased nuclear translocated YAP significantly colocalized with the transcription co-activator molecule pan-TEAD, with a maximum Pearson’s correlation coefficient of 0.51 ± 0.06 at 6 h, compared to YAP-Pan-TEAD colocalization in the WT MHEC, which decreased significantly, with a minimum Pearson’s correlation coefficient of 0.30 ± 0.01 at 6 h. Our data indicate that, during the early phase of infection, upregulated TSP-1 is essential for the regulation of the hippo signaling pathway. These studies advance our understanding of the molecular interactions occurring between heart endothelial cells and T. cruzi, in the presence and absence of TSP-1, providing insights into processes linked to parasite dissemination and pathogenesis.

Thrombospondin-1 expression and modulation of Wnt and hippo signaling pathways during the early phase of Trypanosoma cruzi infection of heart endothelial cells

“…We recently showed that TSP1 plays an important role in the dysregulation of the hippo signaling pathway during the early phase of T . cruzi infection [ 55 ]. In addition, it was suggested that the Wnt/β-catenin pathway is activated early after T .…”

Tankyrase inhibitors hinder Trypanosoma cruzi infection by altering host-cell signalling pathways

“…This early effect might be indicating that Tankyrases are participating in early infection-triggered mechanisms during adhesion, signalling or invasion. In heart endothelial cells, YAP (Yes-associated protein) nuclear levels are altered during the early phase of T. cruzi Infection (Arun et al ., 2020). In immortalized human Schwann cells, T. cruzi infection induces a robust activation of Akt (Chuenkova and PereiraPerrin, 2009).…”

Thrombospondin-1 Signaling Through the Calreticulin/LDL Receptor Related Protein 1 Axis: Functions and Possible Roles in Glaucoma

“…In addition, YAP regulates fibrotic activity of TM cells in response to dexamethasone (Liu et al, 2021). Interestingly, TSP-1 expression is regulated by YAP/TAZ and TSP-1 can also regulate YAP/TAZ signaling (Arun et al, 2020;Yamashiro et al, 2020;Arun et al, 2022). In vascular smooth muscle cells under cyclic mechanical stretch, TSP-1 binds to integrin αvβ1 and induces focal adhesion formation to stimulate nuclear shuttling of YAP, which impacts vascular remodeling in injury models (Yamashiro et al, 2020).…”

“…In vascular smooth muscle cells under cyclic mechanical stretch, TSP-1 binds to integrin αvβ1 and induces focal adhesion formation to stimulate nuclear shuttling of YAP, which impacts vascular remodeling in injury models (Yamashiro et al, 2020). In contrast, heart endothelial cells expressing TSP-1 had lower levels of unphosphorylated YAP and decreased nuclear YAP when challenged with the parasite T. cruzi as compared to endothelial cells from TSP-1 knockout mice (Arun et al, 2020). Furthermore, TSP-1 modulated YAP/β-catenin signaling in this system (Arun et al, 2022).…”