A nanotherapy strategy significantly enhances anticryptosporidial activity of an inhibitor of bifunctional thymidylate synthase-dihydrofolate reductase from Cryptosporidium

Mukerjee, Anindita; Iyidogan, Pinar; Castellanos-González, Alejandro; Cisneros, José A.; Czyzyk, Daniel J.; Ranjan, Amalendu; Jorgensen, William L.; White, A. Clinton; Vishwanatha, Jamboor K.; Anderson, Karen S.

doi:10.1016/j.bmcl.2015.03.091

Cited by 15 publications

(9 citation statements)

References 28 publications

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“…parasite's purine and pyrimidine nucleotide pathways, two of which, dihydrofolate reductase-thymidylate synthase (DHFR-TS) (11) and inosine monophosphate dehydrogenase (IMPDH) (12), are currently pursued as targets for the treatment of cryptosporidiosis. Loss of these enzymes comes without apparent fitness cost, but renders the parasites keenly dependent on host cell nucleotide synthesis.…”

Section: Significancementioning

confidence: 99%

“…The recent advent of CRISPR/Cas9–based genome engineering for Cryptosporidium (10) allowed us to put this hypothesis to a rigorous experimental test. Here we use gene targeting to disrupt multiple enzymes in the parasite’s purine and pyrimidine nucleotide pathways, two of which, dihydrofolate reductase-thymidylate synthase (DHFR-TS) (11) and inosine monophosphate dehydrogenase (IMPDH) (12), are currently pursued as targets for the treatment of cryptosporidiosis. Loss of these enzymes comes without apparent fitness cost, but renders the parasites keenly dependent on host cell nucleotide synthesis.…”

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confidence: 99%

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Genetic ablation of purine salvage in Cryptosporidium parvum reveals nucleotide uptake from the host cell

Pawlowic

Somepalli

Sateriale

et al. 2019

Proc. Natl. Acad. Sci. U.S.A.

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The apicomplexan parasite Cryptosporidium is a leading global cause of severe diarrheal disease and an important contributor to early-childhood mortality. Waterborne outbreaks occur frequently, even in countries with advanced water treatment capabilities, and there is currently no fully effective treatment. Nucleotide pathways are attractive targets for antimicrobial development, and several laboratories are designing inhibitors of these enzymes as potential treatment for Cryptosporidium infections. Here we take advantage of newly available molecular genetics for Cryptosporidium parvum to investigate nucleotide biosynthesis by directed gene ablation. Surprisingly, we found that the parasite tolerates the loss of classical targets including dihydrofolate reductase-thymidylate synthase (DHFR-TS) and inosine monophosphate dehydrogenase (IMPDH). We show that thymidine kinase provides a route to thymidine monophosphate in the absence of DHFR-TS. In contrast, only a single pathway has been identified for C. parvum purine nucleotide salvage. Nonetheless, multiple enzymes in the purine pathway, as well as the adenosine transporter, can be ablated. The resulting mutants are viable under normal conditions but are hypersensitive to inhibition of purine nucleotide synthesis in their host cell. Cryptosporidium might use as-yet undiscovered purine transporters and salvage enzymes; however, genetic and pharmacological experiments led us to conclude that Cryptosporidium imports purine nucleotides from the host cell. The potential for ATP uptake from the host has significant impact on our understanding of parasite energy metabolism given that Cryptosporidium lacks oxidative phosphorylation and glycolytic enzymes are not constitutively expressed throughout the parasite life cycle.

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Section: Significancementioning

confidence: 99%

mentioning

confidence: 99%

Genetic ablation of purine salvage in Cryptosporidium parvum reveals nucleotide uptake from the host cell

Pawlowic

Somepalli

Sateriale

et al. 2019

Proc. Natl. Acad. Sci. U.S.A.

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“…Folate and thymidylate biosynthesis are of vital importance during DNA replication, and dihydrofolate reductase (DHFR) is a key enzyme in folate metabolism, catalyzing the oxidation of NADPH and reduction of dihydrofolate to NADP and tetrahydrofolate [ 69 , 70 ]. The important roles of folate metabolism and nucleic acid synthesis in cryptosporidiosis, mean that currently, inosine monophosphate dehydrogenase (IMPDH) and dihydrofolate reductase-thymidylate synthase (DHFR-TS) are common targets for the treatment of cryptosporidiosis [ 71 , 72 ]. However, the parasite has been found to tolerate the loss of these classical targets, possibly via as-yet-undiscovered purine transporters and salvage enzymes.…”

Section: Discussionmentioning

confidence: 99%

Comparative proteomics reveals Cryptosporidium parvum manipulation of the host cell molecular expression and immune response

Liu

Jiang

et al. 2021

PLoS Negl Trop Dis

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Cryptosporidium is a life-threating protozoan parasite belonging to the phylum Apicomplexa, which mainly causes gastroenteritis in a variety of vertebrate hosts. Currently, there is a re-emergence of Cryptosporidium infection; however, no fully effective drug or vaccine is available to treat Cryptosporidiosis. In the present study, to better understand the detailed interaction between the host and Cryptosporidium parvum, a large-scale label-free proteomics study was conducted to characterize the changes to the proteome induced by C. parvum infection. Among 4406 proteins identified, 121 proteins were identified as differentially abundant (> 1.5-fold cutoff, P < 0.05) in C. parvum infected HCT-8 cells compared with uninfected cells. Among them, 67 proteins were upregulated, and 54 proteins were downregulated at 36 h post infection. Analysis of the differentially abundant proteins revealed an interferon-centered immune response of the host cells against C. parvum infection and extensive inhibition of metabolism-related enzymes in the host cells caused by infection. Several proteins were further verified using quantitative real-time reverse transcription polymerase chain reaction and western blotting. This systematic analysis of the proteomics of C. parvum-infected HCT-8 cells identified a wide range of functional proteins that participate in host anti-parasite immunity or act as potential targets during infection, providing new insights into the molecular mechanism of C. parvum infection.

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“…In the case of anti-Cryptosporidium drugs, there are several structures of protein targets have been resolved using X-Ray diffraction method 31,27,32 . These protein targets are generally derived from several types of enzymes classification, namely dihydrofolate reductase, lactate dehydrogenase, adjacent gene encoding predicted malate dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase, inosine-5'-monophosphate dehydrogenase, bifunctional dihydrofolate reductase-thymidylate synthase and S-adenosylmethionine synthetase [33][34][35][36][37][38][39] . So far, the X-Ray resolutions for all protein structures related to both species of Cryptosporidium have been resolved mostly between 2.0-2.5 Å (angstrom) 28 .…”

Section: Protein Target Selectionmentioning

confidence: 99%

Bioinformatics Applications on Cryptosporidium Research: A Review

Yusof

2022

Bangladesh J Med Sci

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The biology of Cryptosporidium has been studied increasingly since it was recognized as a pathogen of humans more than a century. Its recent recognition as a second leading cause of diarrhoea or cryptosporidiosis immunocompromised patients globally has led many researchers to study on this parasite. Many new technologies such as high-throughput omics and bioinformatics tools have been implemented to investigate this zoonotic parasite in a better approach. The aim of this review article is mainly to briefly describe recent applications of structural bioinformatics in order to reveal the potentiality of a suitable therapeutic target in Cryptosporidium. This review was written based on the search of cited publications in SCOPUS website with the combination of word ‘Cryptosporidium’ with other words like bioinformatics, protein structure, structural biology and homology modeling. The search results then were selected based on the relativeness of updated information needed to be prepared in this review. Several cited publications were used to elaborate the review accordingly despitelimitedreviewupdatesrelatedtoprotein bioinformation of thisparasite. As a conclusion, bioinformatics is a commonly known to be cutting-edge technology that has been recognised for its power to reveal the secret of parasite biology in silico including a neglected parasite, Cryptosporidium. Bangladesh Journal of Medical Science Vol. 21(1) 2022 Page : 8-18

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A nanotherapy strategy significantly enhances anticryptosporidial activity of an inhibitor of bifunctional thymidylate synthase-dihydrofolate reductase from Cryptosporidium

Cited by 15 publications

References 28 publications

Genetic ablation of purine salvage in Cryptosporidium parvum reveals nucleotide uptake from the host cell

Genetic ablation of purine salvage in Cryptosporidium parvum reveals nucleotide uptake from the host cell

Comparative proteomics reveals Cryptosporidium parvum manipulation of the host cell molecular expression and immune response

Bioinformatics Applications on Cryptosporidium Research: A Review

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