Myocardial Chemokine Expression and Intensity of Myocarditis in Chagas Cardiomyopathy Are Controlled by Polymorphisms in CXCL9 and CXCL10

Abstract

Background

Chronic Chagas cardiomyopathy (CCC), a life-threatening inflammatory dilated cardiomyopathy, affects 30% of the approximately 8 million patients infected by Trypanosoma cruzi. Even though the Th1 T cell-rich myocarditis plays a pivotal role in CCC pathogenesis, little is known about the factors controlling inflammatory cell migration to CCC myocardium.

Methods and Results

Using confocal immunofluorescence and quantitative PCR, we studied cell surface staining and gene expression of the CXCR3, CCR4, CCR5, CCR7, CCR8 receptors and their chemokine ligands in myocardial samples from end-stage CCC patients. CCR5+, CXCR3+, CCR4+, CCL5+ and CXCL9+ mononuclear cells were observed in CCC myocardium. mRNA expression of the chemokines CCL5, CXCL9, CXCL10, CCL17, CCL19 and their receptors was upregulated in CCC myocardium. CXCL9 mRNA expression directly correlated with the intensity of myocarditis, as well as with mRNA expression of CXCR3, CCR4, CCR5, CCR7, CCR8 and their ligands. We also analyzed single-nucleotide polymorphisms for genes encoding the most highly expressed chemokines and receptors in a cohort of Chagas disease patients. CCC patients with ventricular dysfunction displayed reduced genotypic frequencies of CXCL9 rs10336 CC, CXCL10 rs3921 GG, and increased CCR5 rs1799988 CC as compared to those without dysfunction. Significantly, myocardial samples from CCC patients carrying the CXCL9/CXCL10 genotypes associated to a lower risk displayed a 2–6 fold reduction in mRNA expression of CXCL9, CXCL10, and other chemokines and receptors, along with reduced intensity of myocarditis, as compared to those with other CXCL9/CXCL10 genotypes.

Conclusions

Results may indicate that genotypes associated to reduced risk in closely linked CXCL9 and CXCL10 genes may modulate local expression of the chemokines themselves, and simultaneously affect myocardial expression of other key chemokines as well as intensity of myocarditis. Taken together our results may suggest that CXCL9 and CXCL10 are master regulators of myocardial inflammatory cell migration, perhaps affecting clinical progression to the life-threatening form of CCC.

Author Summary

Chronic Chagas cardiomyopathy (CCC) is an inflammatory heart disease that affects millions in Latin America, and in growing numbers in USA and Europe. Survival among CCC patients is shorter than among patients with cardiomyopathy of non-inflammatory etiology. This suggests that the inflammatory cell influx plays an important pathogenic role in CCC. However, little is known about the factors that maintain this myocardial inflammation. We hypothesized that Th1 T cell-attracting chemokines, involved in driving leukocyte migration, could play a role in myocardial inflammation. Herein, we have analyzed expression of several chemokines and receptors in heart tissue from patients with CCC and controls. We found inflammatory cells expressing chemokines and receptors consistent with Th1 T cell influx into CCC myocardium. mRNA expression levels of the chemokine CXCL9 correlated with inflammation. We also studied whether genetic variations in these genes could be associated to CCC development. Polymorphisms in CXCL9, CXCL10 and CCR5 were associated to differential risk of progression to the more severe form of CCC. Polymorphisms of CXCL9 and CXCL10 were also associated to the intensity of myocardial inflammation and chemokine expression. These results suggest that such chemokines may be master regulators of myocardial inflammatory cell migration, perhaps affecting clinical progression to severe CCC.