Localized cardiac small molecule trajectories and persistent chemical sequelae in experimental Chagas disease

Abstract

Post-infectious conditions present major health burdens but remain poorly understood. In Chagas disease (CD), caused by Trypanosoma cruzi parasites, antiparasitic agents that successfully clear T. cruzi do not always improve clinical outcomes. In this study, we reveal differential small molecule trajectories between cardiac regions during chronic T. cruzi infection, matching with characteristic CD apical aneurysm sites. Incomplete, region-specific, cardiac small molecule restoration is observed in animals treated with the antiparasitic benznidazole. In contrast, superior restoration of the cardiac small molecule profile is observed for a combination treatment of reduced-dose benznidazole plus an immunotherapy, even with less parasite burden reduction. Overall, these results reveal molecular mechanisms of CD treatment based on simultaneous effects on the pathogen and on host small molecule responses, and expand our understanding of clinical treatment failure in CD. This link between infection and subsequent persistent small molecule perturbation broadens our understanding of infectious disease sequelae.

The impact of vaccine-linked chemotherapy on liver health in a mouse model of chronic Trypanosoma cruzi infection

“…In Liu et al, 2023, we previously reported that combination treatment with low BNZ + vaccine better restored T . cruzi induced metabolic perturbances in several sections of heart compared to curative BNZ treatment [ 30 ]. However, that work did not evaluate the impact of treatments on liver health.…”

“…The T . cruzi H1 strain, transfected with the pTRIX2-RE9h plasmid containing the thermostable, red-shifted firefly luciferase gene PpyRE9h [ 27 – 30 ], was grown on monolayers of the C2C12 mouse myocyte cell line (ATCC CRL-1772) in RPMI media supplemented with 5% fetal bovine serum and 1X Penicillin/Streptomycin (cRPMI) to propagate tissue culture trypomastigotes (TCT). Culture media containing TCT was collected, parasites were pelleted by centrifugation, washed once with sterile medical grade saline, then resuspended in sterile medical grade saline.…”

“…Mice were infected with 5000 trypomastigotes of a bioluminescent clone of the T . cruzi H1 strain, generated in our laboratory [ 30 ], by intraperitoneal injection. Naïve age-matched mice were left uninfected as controls.…”

Small molecule biomarkers predictive of Chagas disease progression

“…Total ion current (TIC) normalization to a constant sum of 1 was performed in Jupyter Notebook using R (version 3.6.1). MZmine output files were uploaded and analyzed by Featurebased molecular networking 47 in GNPS for annotation 33 . Annotations were retrieved using a script developed in our laboratory 48 .…”

“…Specifically, given our prior findings on impacts of infection on glycerophosphocholines and purines 33 , we manually selected m/z 482.361 (lysophosphocholine O-16:0), m/z 137.046 (hypoxanthine), m/z 348.07 (adenosine diphosphate) and m/z 269.088 (xanthosine) to expand the inclusion list. We also included these four candidates for validation analysis in cohort 1 (Table 3).…”