Anti-trypanosomatid drug discovery: progress and challenges

Nature Reviews Microbiology, Год журнала: 2022, Номер 21(1), С. 35 - 50

Опубликована: Авг. 22, 2022

Leishmaniasis (visceral and cutaneous), Chagas disease human African trypanosomiasis cause substantial death morbidity, particularly in low- middle-income countries. Although the situation has improved for trypanosomiasis, there remains an urgent need new medicines to treat leishmaniasis disease; clinical development pipeline is sparse disease. In this Review, we describe recent advances our understanding of biology causative pathogens, from drug discovery perspective, explore progress that been made candidates identification promising molecular targets. We also challenges developing discuss potential solutions overcome such hurdles. Gilbert colleagues kinetoplastid diseases how parasite affects process

Язык: Английский

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Challenges in drug discovery targeting TriTryp diseases with an emphasis on leishmaniasis

International Journal for Parasitology Drugs and Drug Resistance, Год журнала: 2018, Номер 8(3), С. 430 - 439

Опубликована: Сен. 28, 2018

Tritryps diseases are devastating parasitic neglected infections caused by Leishmania spp., Trypanosoma cruzi and brucei subspecies. Together, these parasites affect more than 30 million people worldwide cause high mortality morbidity. Leishmaniasis comprises a complex group of with clinical manifestation ranging from cutaneous lesions to systemic visceral damage. Antimonials, the first-choice drugs used treat leishmaniasis, lead toxicity carry significant contraindications limiting its use. Drug-resistant parasite strains also matter for increasing concern, especially in areas very limited resources. The current scenario calls novel and/or improvement existing therapeutics as key research priorities field. Although several studies have shown advances drug discovery towards leishmaniasis recent years, knowledge gaps pipelines still need be addressed. In this review we discuss not only scientific non-scientific bottlenecks development, but central role public-private partnerships successful campaign treatment options against disease.

Язык: Английский

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Systematic functional analysis of Leishmania protein kinases identifies regulators of differentiation or survival

Nature Communications, Год журнала: 2021, Номер 12(1)

Опубликована: Фев. 23, 2021

Abstract Differentiation between distinct stages is fundamental for the life cycle of intracellular protozoan parasites and transmission hosts, requiring stringent spatial temporal regulation. Here, we apply kinome-wide gene deletion tagging in Leishmania mexicana promastigotes to define protein kinases with transition roles. Whilst 162 are dispensable, 44 kinase genes refractory likely core required parasite replication. Phenotyping pooled mutants using bar-seq projection pursuit clustering reveal functional phenotypic groups involved differentiation from metacyclic promastigote amastigote, growth survival macrophages mice, colonisation sand fly motility. This unbiased interrogation function allows targeted investigation organelle-associated signalling pathways successful parasitism.

Язык: Английский

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The current drug discovery landscape for trypanosomiasis and leishmaniasis: Challenges and strategies to identify drug targets

Drug Development Research, Год журнала: 2020, Номер 83(2), С. 225 - 252

Опубликована: Апрель 6, 2020

Abstract Human trypanosomiasis and leishmaniasis are vector‐borne neglected tropical diseases caused by infection with the protozoan parasites Trypanosoma spp. Leishmania spp., respectively. Once restricted to endemic areas, these now distributed worldwide due human migration, climate change, anthropogenic disturbance, causing significant health economic burden globally. The current chemotherapy used treat has limited efficacy, drug resistance is spreading. Hence, new drugs urgently needed. Phenotypic compound screenings have prevailed as leading method discover candidates against diseases. However, publication of complete genome sequences multiple strains, advances in application CRISPR/Cas9 technology, vivo bioluminescence‐based imaging set stage for advancing target‐based discovery. This review analyses limitations narrow pool available presently treating It describes drug‐based clinical trials highlighting most promising leads. Furthermore, presents a focused discussion on important biological pharmacological challenges that discovery programs must overcome advance candidates. Finally, it examines advantages modern research tools designed identify validate essential genes targets, including genomic editing applications imaging.

Язык: Английский

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A limitation lifted: A conditional knockdown system reveals essential roles for Polo-like kinase and Aurora kinase 1 in Trypanosoma cruzi cell division

Proceedings of the National Academy of Sciences, Год журнала: 2025, Номер 122(8)

Опубликована: Фев. 18, 2025

While advances in genome editing technologies have simplified gene disruption many organisms, the study of essential genes requires development conditional or knockdown systems that are not available most organisms. Such is case for Trypanosoma cruzi, a parasite causes Chagas disease, severely neglected tropical disease endemic to Latin America often fatal. Our knowledge identity and their functions T. cruzi has been constrained by historical challenges very basic genetic manipulation absence RNA interference machinery. Here, we describe use self-cleaving sequences conditionally regulate expression cruzi. Using these tools, identified roles Polo-like Aurora kinases cell division, mirroring brucei. Importantly, demonstrate intracellular amastigotes, disease-causing stage its human host. This system enables efficient scalable functional characterization provides framework other nonmodel

Язык: Английский

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LeishGEdit: A Method for Rapid Gene Knockout and Tagging Using CRISPR-Cas9

Methods in molecular biology, Год журнала: 2019, Номер unknown, С. 189 - 210

Опубликована: Янв. 1, 2019

Язык: Английский

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TransLeish: Identification of membrane transporters essential for survival of intracellular Leishmania parasites in a systematic gene deletion screen

Nature Communications, Год журнала: 2025, Номер 16(1)

Опубликована: Янв. 2, 2025

Abstract For the protozoan parasite Leishmania , completion of its life cycle requires sequential adaptation cellular physiology and nutrient scavenging mechanisms to different environments a sand fly alimentary tract acidic mammalian host cell phagolysosome. Transmembrane transporters are gatekeepers intracellular environments, controlling flux solutes ions across membranes. To discover which vital for survival as amastigote forms, we carried out systematic loss-of-function screen L. mexicana transportome. A total 312 protein components small molecule carriers, ion channels pumps were identified targeted in CRISPR-Cas9 gene deletion promastigote form, yielding 188 viable null mutants. Forty transporter deletions caused significant loss fitness macrophage mouse infections. striking example is Vacuolar H + ATPase (V-ATPase), which, unexpectedly, was dispensable growth vitro but essential disease-causing amastigotes.

Язык: Английский

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Essential roles for deubiquitination in Leishmania life cycle progression

PLoS Pathogens, Год журнала: 2020, Номер 16(6), С. e1008455 - e1008455

Опубликована: Июнь 16, 2020

The parasitic protozoan Leishmania requires proteasomal, autophagic and lysosomal proteolytic pathways to enact the extensive cellular remodelling that occurs during its life cycle. proteasome is essential for parasite proliferation, yet little known about requirement ubiquitination/deubiquitination processes in growth differentiation. Activity-based protein profiling of L. mexicana C12, C19 C65 deubiquitinating cysteine peptidases (DUBs) revealed DUB activity remains relatively constant differentiation procyclic promastigote amastigote. However, when cycle phenotyping (bar-seq) was performed on a pool including 15 barcoded null mutants created promastigotes using CRISPR-Cas9, significant loss fitness observed intracellular infection. DUBs 4, 7, 13 are required successful transformation from metacyclic amastigote 3, 5, 6, 8, 10, 11 14 normal proliferation mice. 1, 2, 12 16 viability role DUB2 establishing infection demonstrated DiCre inducible gene deletion vitro vivo. found nucleus interacts with nuclear proteins associated transcription/chromatin dynamics, mRNA splicing capping. has broad linkage specificity, cleaving all di-ubiquitin chains except Lys27 Met1. Our study demonstrates crucial play survival amastigotes exquisitely sensitive disruption ubiquitination homeostasis.

Язык: Английский

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Need for sustainable approaches in antileishmanial drug discovery

Parasitology Research, Год журнала: 2019, Номер 118(10), С. 2743 - 2752

Опубликована: Авг. 31, 2019

Язык: Английский

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Gene editing and scalable functional genomic screening in Leishmania species using the CRISPR/Cas9 cytosine base editor toolbox LeishBASEedit

eLife, Год журнала: 2023, Номер 12

Опубликована: Май 24, 2023

CRISPR/Cas9 gene editing has revolutionised loss-of-function experiments in Leishmania , the causative agent of leishmaniasis. As lack a functional non-homologous DNA end joining pathway however, obtaining null mutants typically requires additional donor DNA, selection drug resistance-associated edits or time-consuming isolation clones. Genome-wide screens across different conditions and multiple species are therefore unfeasible at present. Here, we report cytosine base editor (CBE) toolbox that overcomes these limitations. We employed CBEs to introduce STOP codons by converting into thymine created http://www.leishbaseedit.net/ for CBE primer design kinetoplastids. Through reporter assays targeting single- multi-copy genes L. mexicana major donovani infantum demonstrate how this tool can efficiently generate expressing just one single-guide RNA, reaching up 100% rate non-clonal populations. then generated -optimised successfully targeted an essential plasmid library delivered screen . Since our method does not require double-strand breaks, homologous recombination, clones, believe enables first time genetic via delivery libraries.

Язык: Английский

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