In Situ Detection of Dormant Trypanosoma cruzi Amastigotes Using Bioluminescent-Fluorescent Reporters

Sánchez-Valdéz, Fernando; Padilla, Ángel M.

doi:10.1007/978-1-4939-9148-8_13

Cited by 7 publications

(5 citation statements)

References 14 publications

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“…Although there have been some new developments in R&D from recent clinical trials, there is an urgent need to review and challenge the current evidence and define clinical research priorities for the immediate future. This would ensure that appropriate, efficacious drugs for CD are developed, but will require a focused and collaborative effort from the entire CD research community [127].…”

Section: Chagas Disease: Key Issues Inadequate R and Dmentioning

confidence: 99%

WHF IASC Roadmap on Chagas Disease

et al. 2020

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Background: Chagas Disease is a neglected tropical disease caused by the protozoan Trypanosoma cruzi, with some of the most serious manifestations affecting the cardiovascular system. It is a chronic, stigmatizing condition, closely associated with poverty and affecting close to 6 million people globally. Although historically the disease was limited to endemic areas of Latin America recent years have seen an increasing global spread. In addition to the morbidity and mortality associated with the disease, the social and economic burdens on individuals and society are substantial. Often called the 'silent killer', Chagas disease is characterized by a long, asymptomatic phase in affected indivi duals. A pproximately 30% then go on develop chronic Chagas cardiomyopathy and other serious cardiac complications such as stroke, rhythm disturbances and severe heart failure. Methods: In a collaboration of the World Hearth Federation (WHF) and the Inter-American S ociety of Cardiology (IASC) a writing group consisting of 20 diverse experts on Chagas disease (CD) was convened. The group provided up to date expert knowledge based on their area of expertise. An extensive review of the literature describing obstacles to diagnosis and treatment Echeverría et al: WHF IASC Roadmap on Chagas Disease Art. 26, page 2 of 31 of CD along with proposed solutions was conducted. A survey was sent to all WHF Members and, using snowball sampling to widen the consultation, to a variety of health care professionals working in the CD global health community. The results were analyzed, open comments were reviewed and consolidated, and the findings were incorporated into this document, thus ensuring a consensus representation. Results: The WHF IASC Roadmap on Chagas Disease offers a comprehensive summary of current knowledge on prevention, diagnosis and management of the disease. In providing an analysis of 'roadblocks' in access to comprehensive care for Chagas disease patients, the document serves as a framework from which strategies for implementation such as national plans can be formulated. Several dimensions are considered in the analysis: healthcare system capabilities, governance, financing, community awareness and advocacy. Conclusion: The WHF IASC Roadmap proposes strategies and evidence-based solutions for healthcare professionals, health authorities and governments to help overcome the barriers to comprehensive care for Chagas disease patients. This roadmap describes an ideal patient care pathway, and explores the roadblocks along the way, offering potential solutions based on available research and examples in practice. It represents a call to action to decision-makers and health care professionals to step up efforts to eradicate Chagas disease.

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Section: Chagas Disease: Key Issues Inadequate R and Dmentioning

confidence: 99%

WHF IASC Roadmap on Chagas Disease

et al. 2020

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“…TH 3.75 mpk suppressed/highly reduced the parasitemia, neither therapeutic scheme was able to reach 100% animal survival and induce sterile cure. Thus, further studies will need to address the efficacy against dormant forms of T. cruzi as recent data suggest the existence of an adaptive difference between parasite strains to generate dormant cells, and that homologous recombination in T. cruzi may be important for dormancy stages (Sańchez-Valdeź and Padilla, 2019;Resende et al, 2020).…”

Section: Combos Meanmentioning

confidence: 99%

Combination With Tomatidine Improves the Potency of Posaconazole Against Trypanosoma cruzi

Rocha-Hasler

Oliveira

Gama

et al. 2021

Front. Cell. Infect. Microbiol.

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Azoles such as posaconazole (Posa) are highly potent against Trypanosoma cruzi. However, when tested in chronic Chagas disease patients, a high rate of relapse after Posa treatment was observed. It appears that inhibition of T. cruzi cytochrome CYP51, the target of azoles, does not deliver sterile cure in monotherapy. Looking for suitable combination partners of azoles, we have selected a set of inhibitors of sterol and sphingolipid biosynthetic enzymes. A small-scale phenotypic screening was conducted in vitro against the proliferative forms of T. cruzi, extracellular epimastigotes and intracellular amastigotes. Against the intracellular, clinically relevant forms, four out of 15 tested compounds presented higher or equal activity as benznidazole (Bz), with EC50 values ≤2.2 μM. Ro48-8071, an inhibitor of lanosterol synthase (ERG7), and the steroidal alkaloid tomatidine (TH), an inhibitor of C-24 sterol methyltransferase (ERG6), exhibited the highest potency and selectivity indices (SI = 12 and 115, respectively). Both were directed to combinatory assays using fixed-ratio protocols with Posa, Bz, and fexinidazole. The combination of TH with Posa displayed a synergistic profile against amastigotes, with a mean ΣFICI value of 0.2. In vivo assays using an acute mouse model of T. cruzi infection demonstrated lack of antiparasitic activity of TH alone in doses ranging from 0.5 to 5 mg/kg. As observed in vitro, the best combo proportion in vivo was the ratio 3 TH:1 Posa. The combination of Posa at 1.25 mpk plus TH at 3.75 mpk displayed suppression of peak parasitemia of 80% and a survival rate of 60% in the acute infection model, as compared to 20% survival for Posa at 1.25 mpk alone and 40% for Posa at 10 mpk alone. These initial results indicate a potential for the combination of posaconazole with tomatidine against T. cruzi.

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“…Recently, novel LUC variants, namely, a red-shifted and a nanoluciferase, with increased luminescence emission have been expressed in T. cruzi (Lewis et al, 2014;Taylor et al, 2019;Ward et al, 2020). Worth noting, these and other LUC-reporter cell lines have attracted more interest for investigating host-parasite interaction and drug response (e.g., relapse to treatment and preclinical assessment of hits) by in vivo imaging (Hyland et al, 2008;Andriani et al, 2011;Henriques et al, 2014;Lewis et al, 2014;Calvet et al, 2017;Silva-Dos-Santos et al, 2017;Sánchez-Valdéz and Padilla, 2019;Taylor et al, 2019;Mann et al, 2020) rather than for in vitro screening of compounds (Bot et al, 2010;Lara et al, 2018;Tayama et al, 2023).…”

Section: Introductionmentioning

confidence: 99%

Development of bioluminescent reporter Trypanosoma cruzi and bioassay for compound screening

Quiroga,

Incerti,

Benitez

et al. 2024

Front. Chem. Biol.

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Trypanosoma cruzi is the etiologic agent of Chagas’ disease, a debilitating and mortal zoonotic illness that threatens the lives of several millions of people in the American continent. The acute phase is underdiagnosed and there is no curative treatment for the chronic stage. This unicellular pathogen colonizes cells from different tissues wherein it replicates and remains hidden from immune recognition. There is an urgent need to develop new chemotherapy, which requires robust screening bioassays against the amastigote, a proliferative and drug-resilient intracellular stage of the parasite. With this aim, here we present the generation and characterization of a bioluminescent reporter cell line of the highly infective strain Dm28c from T. cruzi. Constitutive and stable expression of the reporter gene (a red-shifted luciferase from Photinus pyralis) was achieved in the different developmental stages of the pathogen. The transgene did not affect parasite growth and differentiation. The bioluminescence signal displayed a linear correlation with the parasite number. A 96-well screening assay against the amastigote stage has been established and validated with a small compound library. The bioassay proved sensitive, robust, and amenable for high throughput applications. The reporter cell line may prove highly valuable to address different goals in the early phase of the drug discovery process against American trypanosomiasis.

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In Situ Detection of Dormant Trypanosoma cruzi Amastigotes Using Bioluminescent-Fluorescent Reporters

Cited by 7 publications

References 14 publications

WHF IASC Roadmap on Chagas Disease

WHF IASC Roadmap on Chagas Disease

Combination With Tomatidine Improves the Potency of Posaconazole Against Trypanosoma cruzi

Development of bioluminescent reporter Trypanosoma cruzi and bioassay for compound screening

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