Research Lines

Characterization of cardiac immune mechanisms involved in the host response to infection and wound healing

We are interested in studying the relationship between Trypanosoma cruzi infection and cardiac tissue response. This protozoan parasite grows abundantly in cardiac cells of patients with acute Chagas disease. Still, most patients survive the acute infection and progress to the chronic indeterminate phase with few cardiac lesions. Target cell survival seems critical for the health status of infected individuals since only 30% of them develop chronic cardiopathy. We found that T. cruzi infection protects isolated cardiac cells from apoptosis via the activation of Toll like receptor (TLR)-2 and interleukin (IL)-6 secretion (rev Aoki et al 2012). This protection was improved by pretreatment with cruzipain (the major cysteine protease of T. cruzi) devoid of enzymatic activity. Indeed, cruzipain complexed with chagasin (a parasite cysteine protease inhibitor) triggers the release of IL-6, which acts as an essential anti-apoptotic factor. Strikingly, the cytoprotection inversely correlated with the cysteine protease activity since the survival effect was avoided in the presence of active cruzipain. Through different experimental approaches, we found that cruzipain enzymatically cleaves IL-6 receptor (gp130 subunit), and consequently abrogated its downstream signaling pathways (Ponce et al. 2011). Summing up, our data revealed that the parasite had developed strategies to confer cardiomyocyte protection, but it also modulates the stock of catalytically active cruzipain, which cleaves IL-6 receptor. Since signaling through gp130 by IL-6 family cytokines is involved in multiple steps of the immunological response, this might be a central mechanism for T. cruzi immune evasion.

Growing evidence demonstrates that hypoxia-inducible factor (HIF)-1α plays indispensable roles in driving immune response by triggering the expression of CD73 purinergic ectoenzyme. The purinergic system controls the duration and magnitude of purine signals directed to modulate immune cells by converting extracellular ATP (microbicide/pro-inflammatory) to the immunoregulatory metabolite adenosine by the concerted activity of CD39 and CD73 ecto-enzymes. We have reported that CD73 activity controls macrophage microbicidal responses and increases IL-10-producing T-cell populations in the myocardium of T. cruzi-infected mice. Indeed, the pharmacologic inhibition of CD73 prevents parasite replication in cardiac tissue (Ponce et al. 2016). Interestingly, CD73 activity regulates the anti-parasite immune response in a tissue-dependent manner (Eberhardt et al. 2019). Remarkably, cardiac tissue is susceptible to CD73-generated adenosine.

 

Role of oxidative stress and immuno-metabolism in human Chagas disease

Patients infected with T. cruzi exhibit a substantial reduction in the total peripheral T-cell compartment at the expense of CD8+ T-cells. This T population exhibits increased superficial tyrosine nitration (TN) and impaired cytotoxic functionality. Strikingly, we found that IL-6 prevented the nitration of CD8+ T-cells by diminishing monocyte nitric oxide production in in vitro infected peripheral blood mononuclear cell (PBMCs) cultures. This cytokine exerts a profound anti-oxidative effect in the setting of Chagas disease. In addition, IL-6 increases the frequency of monocytes expressing the ectoenzyme CD39 in cultures of PBMCs from infected patients. We also showed that monocytes from T. cruzi-infected patients exhibit enhanced glycolytic activity, pro-oxidative properties and entail CD8+ T-cell surface nitration in a cell contact-dependent manner. Inhibition of glycolysis in in vitro infected PBMCs decreased the inflammatory properties of monocytes/macrophages and reduced the percentage of TN+CD8+ T-cells, improving their functionality (Sanmarco et al. 2017, 2018, 2019).

Recently, we have reported that the expression of HIF-1α and its gene target CD73 are increased in cardiac leukocytes of patients with end-stage Chagas disease (Eberhardt et al. 2021). Strikingly, the number of immune cells expressing HIF-1α and CD73 correlates with the myocarditis degree and local parasite load. These findings provide evidence that CD73-dependent regulatory pathways are locally triggered in the myocardium of patients with end-stage Chagas disease.

 

Optimization of new multiparticulate drug delivery systems for Chagas disease therapy

The development of innovative strategies employing new drug-controlled release systems is an attractive alternative to be considered to improve Chagas disease therapy. In this regard, a special interest has been focused on the development of multiparticulate drug delivery systems (MDDS), which involve coated drug particles or drug in a matrix based on one or more carrier polymers to control or sustain the release of the drug into circulation or into a given target tissues. In this context, several kinds of oppositely charged polyelectrolytes that interact electrostatically in aqueous media and form soluble or insoluble interpolyelectrolyte complexes have been developed. These complexes exhibit higher capabilities to modulate the rate of drug release and improved delivery behavior in comparison to homologous binary complexes, composed by a single polymer, showing remarkable robustness in front of pH changes in the release media.

Benznidazole (BZ) is first-line drug for the treatment of Chagas disease; however, it presents several disadvantages that could hamper its therapeutic success. In collaboration with Dr. Mónica García (UNITEFA-CONICET), we have recently reported a comprehensive biopharmaceutical characterization of different MDDS as carriers of BZ constituted by interpolyelectrolyte complexes based on Eudragit EPO-Eudragit L100 loaded with BZ (Garcia et al. 2018). Employing standardized protocols, we are studying the efficacy and safety of BZ-MDDS for the treatment of Chagas disease. Preliminary results evidence that BZ-MDDS could be a promising alternative to improve pharmacological Chagas disease treatment.

Publications

Special Issue “Trypanosoma cruzi infection: pathogenesis and treatment”, para el Journal Biochimica et Biophysica Acta (BBA) Molecular Basis of Diseases. - Guess Editor Aoki MP; Co-Editor Dr. Juan Bustamante (University of Georgia) 13 scientific articles.

https://www.sciencedirect.com/journal/biochimica-et-biophysica-acta-bba-molecular-basis-of-disease/special-issue/102GT7FVJ4S

Book chapter: Purification and phenotypic characterization of inflammatory cells from clinical samples: Heart Macrophages. Eberhardt, N.; Sanmarco, LM; Aoki, MP.

Methods in Molecular Biology. 2019; 1955:381-395 Editorial Springer Nature.

doi: 10.1007/978-1-4939-9148-8_28

 

PubMed My Bibliography:

  https://www.ncbi.nlm.nih.gov/myncbi/1lo2w48mL6oYV5/bibliography/public/

  1. Aoki MP, Bustamante JM. Preface. Biochim Biophys Acta Mol Basis Dis. 2020 Dec 1;1866(12):165953. doi: 10.1016/j.bbadis.2020.165953.
  2. Eberhardt N; Sanmarco LM; Bergero G; Favaloro RR; Vigliano C; Aoki MP. HIF-1α and CD73 expression in cardiac leukocytes correlates with the severity of myocarditis in end-stage Chagas disease patients. Journal of Leukocyte Biology (2021) Jan;109(1):233-244. Epub 2020 May 25. doi: 10.1002/JLB.4MA0420-125R
  3. Eberhardt N; Sanmarco LM; Bergero G; Theumer MG; García MC; Ponce NE; Cano RC; Aoki MP. Deficiency of CD73 activity promotes protective cardiac immunity against Trypanosoma cruzi infection but permissive environment in visceral adipose tissue. BBA Molecular Basis of Diseases (2020). https://doi.org/10.1016/j.bbadis.2019.165592. IF: 4.71
  4. Sanmarco LM, Eberhardt N, Bergero G, Quebrada Palacio LP, Martino Adami P, Visconti LM, Minguez RA, Hernández-Vasquez Y, Carrera Silva EA, Morelli L, Postan M, Aoki MP. Monocyte glycolysis determines T-cells functionality in human Chagas Disease. JCI Insight. (2019);4 18:e123490. https://doi.org/10.1172/jci.insight.123490. IF: 6.19
  5. Raimondo N; Butassi E; Nicolas JC; Asenza P; Martins M; Cassinerio A; Martinez Wassaf M; Aoki MP. Evaluación de niveles séricos de IL-6, PCR-hs, dímero D y óxido nítrico como marcadores pronóstico de comorbilidades no asociadas a SIDA. Bitácora Digital. (2020): Vol. 6 Núm. 10.
  6. Nowak W, Grendas LN, Sanmarco LM; Estecho, IG; Arena AR; Eberhardt N; Rodante DE; Aoki MP; Daray, FM; Carrera Silva EA; Errasti AE. Pro-inflammatory monocyte profile in patients with major depressive disorder and the induction of M2 macrophage program by ketamine via NMDAR and mTOR. EBioMedicine (2019). https://doi:10.1016/j.ebiom.2019.10.063. IF:78
  7. Carabelli, J; Prato, C; Sanmarco, LM; Aoki, MP; Campetella, O; Tribulatti, M. Interleukin-6 signaling mediates Galectin-8 costimulatory activity of antigen-specific CD4 T cell response. Immunology (2018) jul 4. doi: 10.1111/imm.12980 both authors contributed equally. IF: 3.358.
  8. García MC, Martinelli M; Ponce NE; Sanmarco LM; Aoki MP; Manzo RH; Jimenez-Kairuz AF. Multi-kinetic release of benznidazole-loaded multiparticulate drug delivery systems based on polymethacrylate interpolyelectrolyte complexes. European Journal of Pharmaceutical Sciences (2018) 120 107–122. https://doi.org/10.1016/j.ejps.20104.034. IF: 3.532.
  9. Sanmarco LM; Eberhardt N; Ponce NE; Cano RC; Bonacci G; Aoki MP. New insights into the immunobiology of mononuclear phagocytic cells and their relevance to the pathogenesis of cardiovascular diseases. Revision article. Frontiers in Immunology (2018) Jan 9; 8:1921. DOI: https://www.frontiersin.org/articles/10.3389/fimmu.2017.01921/full. IF: 5.511.
  10. Sanmarco LM; Ponce NE; Visconti LM; Eberhardt N; Theumer MG; Minguez AR; Aoki MP. IL-6 promotes M2 macrophage polarization by modulating purinergic signaling and regulates the lethal release of nitric oxide during Trypanosoma cruzi infection. Biochimica et Biophysica Acta Molecular Basis of Disease (2017) Apr;1863(4):857-869. DOI: 1016/j.bbadis.2017.01.006. IF: 5.108.
  11. Sanmarco LM; Visconti LV; Ramello MC; Ponce NE; Spitale NB, Vozza ML; Bernardi A; Gea S; Minguez AR; Aoki MP. IL-6 improves the nitric oxide induced-cytotoxic CD8+ T cell dysfunction in human Chagas disease. Frontiers in Immunology (2016) 23;7:626. DOI: 10.3389/fimmu.2016.00626. IF: 6.732.
  12. Ponce NE; Sanmarco LM; Eberhardt N; Garcia MC; Rivarola W; Cano RC; Aoki MP. CD73 inhibition shifts cardiac macrophage polarization toward a microbicidal phenotype and ameliorates the outcome of experimental Chagas cardiomyopathy. Journal of Immunology (2016) Aug 1;197(3):814-23. doi: 10.4049/jimmunol.1600371. IF: 4.985.
  13. Garcia, MC; Ponce, NE; Sanmarco, LM; Manzo, RH; Jimenez-Kairuz, AF; Aoki, MP. Clomipramine and Benznidazole act synergistically and ameliorate the outcome of experimental Chagas disease: In vivo and in vitro assessments. Antimicrobial Agents and Chemotherapy (2016) May 23;60(6):3700-8. doi: 10.1128/AAC.00404-16. IF: 4.415
  14. Cabalén, ME; Cabral, M; Sanmarco, LM; Andrada, M; Onofrio, LI; Ponce, NE; Aoki, MP; Gea, S; Cano, RC. Chronic Trypanosoma cruzi infection potentiates adipose tissue macrophage polarization toward an anti-inflammatory M2 phenotype and contributes to diabetes progression in a diet-induced obesity model. Oncotarget (2016) Mar 22;7(12):13400-15. doi: 10.18632/oncotarget.7630. IF: 5.168
  15. Arocena RA, Onofrio IL, Pellegrini VA, Carrera Silva EA, Paroli A, Cano CR, Aoki MP, Gea S. Myeloid-derived suppressor cells are key players in the resolution of inflammation during a model of acute infection. European Journal of Immunology (2014); 44(1):184-94. DOI: 10.1002/eji.201343606. IF: 4.78
  16. Ponce NE; Carrera-Silva EA; Pellegrini AV, Cazorla SI, Malchiodi LE; Lima AP; S Gea; Aoki MP. Trypanosoma cruzi, the causative agent of Chagas disease, modulates IL6-induced STAT3 phosphorylation via gp-130 cleavage in different host cells. Biochimica et Biophysica Acta (2013) 1832 485-494. http://dx.doi.org/10.1016/j.bbadis.2012.12.003. IF: 5.38.
  17. Ponce N.E.; Gea S.; Sanmarco L.M.; Aoki M.P. El Trypanosoma cruzi, agente causal de la enfermedad de Chagas, modula la señalización inducida por interleuquina-6 a través de la degradación del receptor gp130 en diferentes células del huésped. Revista Bitácor@ digital (2013) 3(1) 1-4. ISSN: 2344-9144. Revista Científico-Académica de la Facultad de Ciencias Químicas (UNC). http://revistas.unc.edu.ar/index.php/Bitacora/article/view/6318
  18. Ponce NE; Cano RC; Carrera-Silva EA; Lima AP; Gea S; Aoki MP. Toll-like receptor-2 and Interleukine-6 mediate cardiomyocyte protection from apoptosis during Trypanosoma cruzi murine Infection. Medical Microbiology and Immunology (2012); 201: 145-155. Publicado online 2011 Octubre 9. DOI: 10.1007/s00430-011-0216-z. IF: 3,83.
  19. Aoki MP, Carrera-Silva EA; Cuervo H, Fresno M, Gironés N, Gea S. Non-immune cells contribute to cross-talk between immune cells and inflammatory mediators in the innate response to Trypanosoma cruzi infection.. Revision Article. Journal of Parasitology Research (2012), Article ID 737324 DOI 10.1155/2012/737324.
  20. Savino W, Villa-Verde SM, Mendes-da-Cruz D, Silva-Monteiro E, Perez AR, Aoki MP, Bottasso O, Guiñazú N, Silva-Barbosa SD and Gea S. Cytokines and cell adhesion receptors in the regulation of immunity to Trypanosoma cruzi. Cytokine & Growth Factor Reviews (2007) 18: 107-124. Revisión. DOI:10.1016/j.cytogfr.2007.01.010. IF: 11,82

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