3D mapping of host-parasite-microbiome interactions reveals metabolic determinants of tissue tropism and disease tolerance in Chagas disease

AbstractChagas disease (CD) is a parasitic infection caused by Trypanosoma cruzi protozoa. Over 8 million people worldwide are T. cruzi-positive, 20-30% of which will develop cardiomyopathy, megaoesophagus and/or megacolon. The mechanisms leading to gastrointestinal (GI) symptom development are however poorly understood. To address this issue, we systematically characterized the spatial impact of experimental T. cruzi infection on the microbiome and metabolome across the GI tract. The largest microbiota perturbations were observed in the proximal large intestine in both acute and chronic disease, with chronic-stage effects also observed in the cecum. Strikingly, metabolomic impact of acute-to-chronic stage transition differed depending on the organ, with persistent large-scale effects of infection primarily in the oesophagus and large intestine, providing a potential mechanism for GI pathology tropism in CD. Infection particularly affected acylcarnitine and lipid metabolism. Building on these observations, treatment of infected mice with carnitine-supplemented drinking water prevented acute-stage mortality with no changes in parasite burden. Overall, these results identified a new mechanism of disease tolerance in CD, with potential for the development of new therapeutic regimens. More broadly, these results highlight the potential of spatially-resolved metabolomic approaches to provide insight into disease pathogenesis, with translational applications for infectious disease drug development.

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Mapping of host-parasite-microbiome interactions reveals metabolic determinants of tropism and tolerance in Chagas disease

Science Advances ◽

10.1126/sciadv.aaz2015 ◽

2020 ◽

Vol 6 (30) ◽

pp. eaaz2015 ◽

Cited By ~ 2

Author(s):

E. Hossain ◽

S. Khanam ◽

D. A. Dean ◽

C. Wu ◽

S. Lostracco-Johnson ◽

...

Keyword(s):

Chagas Disease ◽

Large Scale ◽

Scale Effects ◽

Parasite Burden ◽

Acute Stage ◽

Metabolic Disturbances ◽

Spatially Resolved ◽

Cardiac Strain ◽

Host Parasite ◽

The Impact

Chagas disease (CD) is a parasitic disease caused by Trypanosoma cruzi protozoa, presenting with cardiomyopathy, megaesophagus, and/or megacolon. To determine the mechanisms of gastrointestinal (GI) CD tissue tropism, we systematically characterized the spatial localization of infection-induced metabolic and microbiome alterations, in a mouse model of CD. Notably, the impact of the transition between acute and persistent infection differed between tissue sites, with sustained large-scale effects of infection in the esophagus and large intestine, providing a potential mechanism for the tropism of CD within the GI tract. Infection affected acylcarnitine metabolism; carnitine supplementation prevented acute-stage CD mortality without affecting parasite burden by mitigating infection-induced metabolic disturbances and reducing cardiac strain. Overall, results identified a previously-unknown mechanism of disease tolerance in CD, with potential for new therapeutic regimen development. More broadly, results highlight the potential of spatially resolved metabolomics to provide insight into disease pathogenesis and infectious disease drug development.

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A systematic review of historical and current trends in Chagas disease

Therapeutic Advances in Infectious Disease ◽

10.1177/20499361211033715 ◽

2021 ◽

Vol 8 ◽

pp. 204993612110337

Author(s):

Diego-Abelardo Álvarez-Hernández ◽

Rodolfo García-Rodríguez-Arana ◽

Alejandro Ortiz-Hernández ◽

Mariana Álvarez-Sánchez ◽

Meng Wu ◽

...

Keyword(s):

Systematic Review ◽

Chagas Disease ◽

Case Reports ◽

Clinical Manifestations ◽

Human Case ◽

Acute Stage ◽

Diagnostic Tools ◽

Chronic Stage ◽

Pubmed Database ◽

Trypanocidal Drugs

Introduction: Chagas disease (CD) is caused by Trypanosoma cruzi. When acquired, the disease develops in stages. For diagnosis, laboratory confirmation is required, and an extensive assessment of the patient’s health should be performed. Treatment consists of the administration of trypanocidal drugs, which may cause severe adverse effects. The objective of our systematic review was to analyze data contained in the CD published case reports to understand the challenges that patients and clinicians face worldwide. Materials and methods: We performed a systematic review following the PRISMA guidance. PubMed database was explored using the terms ‘American trypanosomiasis’ or ‘Chagas disease’. Results were limited to human case reports written in English or Spanish. A total of 258 reports (322 patients) were included in the analysis. Metadata was obtained from each article. Following this, it was analyzed to obtain descriptive measures. Results: From the sample, 56.2% were males and 43.8% were females. Most cases were from endemic countries (85.4%). The most common clinical manifestations were fever during the acute stage (70.0%), dyspnea during the chronic stage in its cardiac form (53.7%), and constipation during the chronic stage in its digestive form (73.7%). Most patients were diagnosed in the chronic stage (72.0%). Treatment was administered in 56.2% of cases. The mortality rate for the acute stage cases was 24.4%, while for the chronic stage this was 28.4%. Discussion: CD is a parasitic disease endemic to Latin America, with increasing importance due to human and vector migration. In this review, we report reasons for delays in diagnosis and treatment, and trends in medical practices. Community awareness must be increased to improve CD’s diagnoses; health professionals should be appropriately trained to detect and treat infected individuals. Furthermore, public health policies are needed to increase the availability of screening and diagnostic tools, trypanocidal drugs, and, eventually, vaccines.

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Limited Ability of Posaconazole To Cure both Acute and Chronic Trypanosoma cruzi Infections Revealed by Highly SensitiveIn VivoImaging

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00520-15 ◽

2015 ◽

Vol 59 (8) ◽

pp. 4653-4661 ◽

Cited By ~ 84

Author(s):

Amanda Fortes Francisco ◽

Michael D. Lewis ◽

Shiromani Jayawardhana ◽

Martin C. Taylor ◽

Eric Chatelain ◽

...

Keyword(s):

Clinical Trial ◽

Trypanosoma Cruzi ◽

Chagas Disease ◽

Imaging System ◽

Ex Vivo ◽

Parasite Burden ◽

Significant Activity ◽

Content Type ◽

Chronic Stage

ABSTRACTThe antifungal drug posaconazole has shown significant activity againstTrypanosoma cruziin vitroand in experimental murine models. Despite this, in a recent clinical trial it displayed limited curative potential. Drug testing is problematic in experimental Chagas disease because of difficulties in demonstrating sterile cure, particularly during the chronic stage of infection when parasite burden is extremely low and tissue distribution is ill defined. To better assess posaconazole efficacy against acute and chronic Chagas disease, we have exploited a highly sensitive bioluminescence imaging system which generates data with greater accuracy than other methods, including PCR-based approaches. Mice inoculated with bioluminescentT. cruziwere assessed byin vivoandex vivoimaging, with cyclophosphamide-induced immunosuppression used to enhance the detection of relapse. Posaconazole was found to be significantly inferior to benznidazole as a treatment for both acute and chronicT. cruziinfections. Whereas 20 days treatment with benznidazole was 100% successful in achieving sterile cure, posaconazole failed in almost all cases. Treatment of chronic infections with posaconazole did however significantly reduce infection-induced splenomegaly, even in the absence of parasitological cure. The imaging-based screening system also revealed that adipose tissue is a major site of recrudescence in mice treated with posaconazole in the acute, but not the chronic stage of infection. Thisin vivoscreening model for Chagas disease is predictive, reproducible and adaptable to diverse treatment schedules. It should provide greater assurance that drugs are not advanced prematurely into clinical trial.

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Increased heart fibrosis and acute infection in a murine Chagas disease model associated with organophosphorus pesticide metabolite exposure

Scientific Reports ◽

10.1038/s41598-019-54218-7 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 1

Author(s):

Dunia Margarita Medina-Buelvas ◽

Elizabet Estrada-Muñiz ◽

Miriam Rodríguez-Sosa ◽

Mineko Shibayama ◽

Libia Vega

Keyword(s):

Chagas Disease ◽

Cardiac Fibrosis ◽

Acute Infection ◽

Disease Model ◽

Parasitic Infection ◽

Organophosphorus Pesticide ◽

Parasite Burden ◽

Myocardial Damage ◽

Inflammatory Cells ◽

Inflammatory Macrophages

AbstractSome reports suggest that exposure to organophosphorus (OP) pesticides increases the incidence of infections. Ethylated dialkylphosphates (EtDAPs) are metabolites of OP pesticides widely distributed with immunomodulatory potential. Chagas disease is produced by Trypanosoma cruzi parasites, and resolution of this infection requires the activation of inflammatory macrophages (MΦ), which results in cardiac fibrosis. Some reports indicate that EtDAPs increase the amount of the anti-inflammatory alternatively activated MΦ (M2; CD206+F4/80+). Therefore, we analyzed the course of T. cruzi infection, MΦ profiles from peritoneal exudate cells (PECs), inflammatory cell infiltration and fibrosis in the heart of BALB/c mice exposed to diethyldithiophosphate (DEDTP), diethylthiophosphate (DETP) or diethylphosphate (DEP, 0.01 g/kg), common DAPs produced by OP pesticides, 24 h before infection with T. cruzi. We found that DEDTP increased the parasite burden in blood by 99% at the peak of the infection and enhanced the myocardial damage due to an increase in infiltrated inflammatory cells (induced by DEDTP or DETP) and fibrosis (induced by EtDAPs). In the PECs, exposure to EtDAPs increased the proportion of the MΦ subpopulations of M2a, M2b and M2d, which are associated with tissue repair. These results indicate that exposure to EtDAPs can exacerbate the acute phase of a parasitic infection and increase the long-term damage to the heart.

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Tolerance of Benznidazole in Treatment of Chagas' Disease in Adults

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00537-10 ◽

2010 ◽

Vol 54 (11) ◽

pp. 4896-4899 ◽

Cited By ~ 114

Author(s):

María-Jesús Pinazo ◽

José Muñoz ◽

Elizabeth Posada ◽

Paulo López-Chejade ◽

Montserrat Gállego ◽

...

Keyword(s):

Public Health ◽

Side Effects ◽

Trypanosoma Cruzi ◽

European Country ◽

Chagas Disease ◽

Health Problem ◽

Public Health Problem ◽

Acute Stage ◽

Chronic Stage

ABSTRACT Chagas’ disease is an emerging public health problem in areas where the disease is not endemic. Treatment with benznidazole has shown efficacy in the acute stage of the disease, but its efficacy in the chronic stage remains controversial, and unwanted side effects are more frequent and severe in adults than in children. This study describes the profile of side effects of benznidazole in a cohort of Trypanosoma cruzi-infected patients in a European country.

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A longitudinal study of white matter functional network in mild traumatic brain injury

10.1101/2020.09.25.313338 ◽

2020 ◽

Author(s):

Xiaoyan Jia ◽

Xuebin Chang ◽

Lijun Bai ◽

Yulin Wang ◽

Debo Dong ◽

...

Keyword(s):

Traumatic Brain Injury ◽

Brain Injury ◽

White Matter ◽

Mild Traumatic Brain Injury ◽

Cognitive Deficits ◽

Large Scale ◽

Acute Stage ◽

Functional Networks ◽

Long Distance ◽

Chronic Stage

AbstractThe mild traumatic brain injury (mTBI) results in traumatic axonal injury, which damages the long-distance white matter (WM) connections and thus disrupts the functional connectome of large-scale brain networks that support cognitive function. Patterns of WM structural damage following mTBI were well documented using diffusion tensor imaging, however, the functional organization of WM and its association with grey matter functional networks (GM-FNs) and cognitive assessments remains unknown. The present study adopted resting-state functional magnetic resonance imaging to explore WM functional properties in mTBI patients (113 acute patients, 56 chronic patients, 47 healthy controls (HCs)). Eleven large-scale WM functional networks (WM-FNs) were constructed by the k-means clustering algorithm which carried out in voxel-wise WM functional connectivity (FC). Compared to HCs, acute mTBI patients showed enhanced FC between inferior fronto-occipital fasciculus (IFOF) WM-FN and primary sensorimotor WM-FNs, and cortical primary sensorimotor GM-FNs. And FC between IFOF WM-FN and anterior cerebellar GM-FN was positively correlated with information processing speed. Moreover, all of these WM-FNs abnormalities were returned to the normal level at the chronic stage. Our findings suggest the compensatory mechanism of cognitive deficits in the acute stage and its involvement in facilitating recovery from cognitive deficits in the chronic stage. The convergent damage of the IFOF network highlighted its key role in our understanding of the pathophysiology mechanism of mTBI patients and thus might be regarded as a biomarker in the acute stage and a potential indicator of treatment effect.

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Spatial metabolomics identifies localized chemical changes in heart tissue during chronic cardiac Chagas disease

10.1101/2020.06.29.178038 ◽

2020 ◽

Author(s):

Danya A. Dean ◽

Gautham ◽

Jair L. Siqueira-Neto ◽

James H. McKerrow ◽

Pieter C. Dorrestein ◽

...

Keyword(s):

Trypanosoma Cruzi ◽

Chagas Disease ◽

Parasite Burden ◽

Heart Tissue ◽

Blood Transfusions ◽

Organ Transplants ◽

Chronic Stage ◽

Congenital Transmission ◽

Cardiac Symptoms ◽

The Impact

AbstractChagas disease (CD) is one of thirteen neglected tropical diseases caused by the parasite Trypanosoma cruzi. CD is a vector-borne disease transmitted by triatomines but CD can also be transmitted through blood transfusions, organ transplants and congenital transmission. While endemic to Latin America, T. cruzi infects 7-8 million people worldwide and can induce severe cardiac symptoms including apical aneurysms, thromboembolisms and arrhythmias during the chronic stage of CD. However, these cardiac clinical manifestations and CD disease pathogenesis are not fully understood. Using spatial metabolomics (chemical cartography), we sought to understand the localized impact of infection on the cardiac metabolome of mice chronically infected with two divergent T. cruzi strains. Our data showed chemical differences in localized cardiac regions upon chronic T. cruzi infection, indicating that parasite infection changes the host metabolome at select sites in chronic CD. These sites were distinct from the sites of highest parasite burden. In addition, we identified acylcarnitines and phosphocholines as discriminatory chemical families within each heart region, comparing infected and uninfected samples. Overall, our study indicated overall and positional metabolic differences common to infection with different T. cruzi strains, and identified select infection-modulated pathways. These results provide further insight into CD pathogenesis and demonstrate the advantage of a spatial perspective to understand infectious disease tropism.Author SummaryChagas disease (CD) is a tropical disease caused by the parasite Trypanosoma cruzi. CD originated in South America; however, there are now 7-8 million people infected worldwide due to population movements. CD is transmitted through a triatomine vector, organ transplants, blood transfusions and congenital transmission. It occurs in two stages, an acute stage (usually asymptomatic) and the chronic stage. Chronic stage CD presents with severe cardiac symptoms such as heart failure, localized aneurysms and cardiomyopathy. Unfortunately, what causes severe cardiac symptoms in some individuals in chronic CD is not fully understood. Therefore, we used liquid chromatography-tandem mass spectrometry to analyze the heart tissue of chronically T. cruzi-infected and uninfected mice, to understand the impact of infection on the tissue metabolome. We identified discriminatory small molecules related to T. cruzi infection. We also determined that regions with the highest parasite burden are distinct from the regions with the largest changes in overall metabolite profile; these locations of high metabolic perturbation provide a molecular mechanism to why localized cardiac symptoms occur in CD. Overall, our work gives insight to chronic cardiac CD symptom development and shapes a framework for novel treatment and biomarker development.

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Image-Based In Vitro Screening Reveals the Trypanostatic Activity of Hydroxymethylnitrofurazone against Trypanosoma cruzi

International Journal of Molecular Sciences ◽

10.3390/ijms22136930 ◽

2021 ◽

Vol 22 (13) ◽

pp. 6930

Author(s):

Cauê Benito Scarim ◽

Francisco Olmo ◽

Elizabeth Igne Ferreira ◽

Chung Man Chin ◽

John M. Kelly ◽

...

Keyword(s):

Trypanosoma Cruzi ◽

Chagas Disease ◽

Ex Vivo ◽

Therapeutic Option ◽

Acute Stage ◽

Reporter Strain ◽

Fluorescent Reporter ◽

Chronic Stage

Hydroxymethylnitrofurazone (NFOH) is a therapeutic candidate for Chagas disease (CD). It has negligible hepatotoxicity in a murine model compared to the front-line drug benznidazole (BZN). Here, using Trypanosoma cruzi strains that express bioluminescent and/or fluorescent reporter proteins, we further investigated the in vitro and in vivo activity of NFOH to define whether the compound is trypanocidal or trypanostatic. The in vitro activity was assessed by exploiting the fluorescent reporter strain using wash-out assays and real-time microscopy. For animal experimentation, BALB/c mice were inoculated with the bioluminescent reporter strain and assessed by highly sensitive in vivo and ex vivo imaging. Cyclophosphamide treatment was used to promote parasite relapse in the chronic stage of infection. Our data show that NFOH acts by a trypanostatic mechanism, and that it is more active than BZN in vitro against the infectious trypomastigote form of Trypanosoma cruzi. We also found that it is more effective at curing experimental infections in the chronic stage, compared with the acute stage, a feature that it shares with BZN. Therefore, given its reduced toxicity, enhanced anti-trypomastigote activity, and curative properties, NFOH can be considered as a potential therapeutic option for Chagas disease, perhaps in combination with other trypanocidal agents.

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Assessing the Effectiveness of Curative Benznidazole Treatment in Preventing Chronic Cardiac Pathology in Experimental Models of Chagas Disease

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00832-18 ◽

2018 ◽

Vol 62 (10) ◽

Cited By ~ 9

Author(s):

Amanda Fortes Francisco ◽

Shiromani Jayawardhana ◽

Martin C. Taylor ◽

Michael D. Lewis ◽

John M. Kelly

Keyword(s):

Chagas Disease ◽

Cardiac Fibrosis ◽

Early Stage ◽

Population Level ◽

Heart Tissue ◽

Experimental Models ◽

Acute Stage ◽

Cardiac Pathology ◽

Content Type ◽

Chronic Stage

ABSTRACT Chagasic heart disease develops in 30% of those infected with the protozoan parasite Trypanosoma cruzi, but can take decades to become symptomatic. Because of this, it has been difficult to assess the extent to which antiparasitic therapy can prevent the development of pathology. We sought to address this question using experimental murine models, exploiting highly sensitive bioluminescent imaging to monitor curative efficacy. Mice were inoculated with bioluminescent parasites and then cured in either the acute or chronic stage of infection with benznidazole. At the experimental endpoint (5 to 6 months postinfection), heart tissue was removed and assessed for inflammation and fibrosis, two widely used markers of cardiac pathology. Infection of BALB/c and C3H/HeN mice with distinct T. cruzi lineages resulted in greatly increased myocardial collagen content at a group level, indicative of fibrotic pathology. When mice were cured by benznidazole in the acute stage, the development of pathology was completely blocked. However, if treatment was delayed until the chronic stage, cardiac fibrosis was observed in the BALB/c model, although the protective effect was maintained in the case of C3H/HeN mice. These experiments therefore demonstrate that curative benznidazole treatment early in murine T. cruzi infections can prevent the development of cardiac fibrosis. They also show that treatment during the chronic stage can block pathology but the effectiveness varies between infection models. If these findings are extendable to humans, it implies that widespread chemotherapeutic intervention targeted at early-stage infections could play a crucial role in reducing Chagas disease morbidity at a population level.

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Immunity conferred by drug-cured experimental Trypanosoma cruzi infections is long-lasting and cross-strain protective

10.1101/741462 ◽

2019 ◽

Author(s):

Gurdip Singh Mann ◽

Amanda F. Francisco ◽

Shiromani Jayawardhana ◽

Martin C. Taylor ◽

Michael D. Lewis ◽

...

Keyword(s):

Trypanosoma Cruzi ◽

Chagas Disease ◽

Bioluminescence Imaging ◽

Viral Vector ◽

Parasite Burden ◽

Whole Body ◽

Luciferase Reporter ◽

Chronic Stage ◽

Route Of Infection ◽

Highly Sensitive

ABSTRACTBackgroundThe long term and complex nature of Chagas disease in humans has restricted studies on vaccine feasibility. Animal models also have limitations due to technical difficulties in monitoring the extremely low parasite burden that is characteristic of chronic stage infections. Advances in imaging technology offer alternative approaches that circumvent these problems. Here, we describe the use of highly sensitive whole body in vivo imaging to assess the efficacy of recombinant viral vector vaccines and benznidazole-cured infections to protect mice from challenge with Trypanosoma cruzi.Methodology/Principal FindingsMice were infected with T. cruzi strains modified to express a red-shifted luciferase reporter. Using bioluminescence imaging, we assessed the degree of immunity to re-infection conferred after benznidazole-cure. Mice infected for 14 days or more, prior to the initiation of treatment, were highly protected from challenge with both homologous and heterologous strains (>99% reduction in parasite burden). Sterile protection against homologous challenge was frequently observed. This level of protection was considerably greater than that achieved with recombinant vaccines. It was also independent of the route of infection or size of the challenge inoculum, and was long-lasting, with no significant diminution in immunity after almost a year. When the primary infection was benznidazole-treated after 4 days (before completion of the first cycle of intracellular infection), the degree of protection was much reduced, an outcome associated with a minimal T. cruzi-specific IFN-γ+ T cell response.Conclusions/SignificanceOur findings suggest that a protective Chagas disease vaccine must have the ability to eliminate parasites before they reach organs/tissues, such as the GI tract, where once established, they become largely refractory to the induced immune response.AUTHOR SUMMARYChagas disease, which is caused by the protozoan parasite Trypanosoma cruzi, is a major public health problem throughout Latin America. Attempts to develop a vaccine have been hampered by technical difficulties in monitoring the extremely low parasite burden during the life-long chronic stage of infection. To circumvent these issues, we used highly sensitive bioluminescence imaging to assess the ability of recombinant viral vector vaccines and drug-cured infections to confer protection against experimental challenge in mice. We observed that drug-cured infections were much more effective than subunit vaccines, with many instances of sterile protection. Efficacy was independent of the route of infection or size of the challenge inoculum, and was undiminished after almost a year. In addition, drug-cured infections conferred a high level of cross-strain protection. The highly sensitive imaging procedures enabled us to visualise parasite distribution in mice where sterile protection was not achieved. This suggested that to confer sterile protection, vaccines must prevent the infection of organs/tissues that act as parasite reservoirs during the chronic stage. Once established at these sites, parasites become largely refractory to vaccine-induced elimination.

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