A Target Repurposing Approach Identifies N-myristoyltransferase as a New Candidate Drug Target in Filarial Nematodes

Galvin, Brendan D.; Li, Zhi‐Ru; Villemaine, Estelle; Poole, Catherine B.; Chapman, Melissa; Pollastri, Michael P.; Wyatt, Paul G.; Carlow, Clotilde K. S.

doi:10.1371/journal.pntd.0003145

Cited by 21 publications

(13 citation statements)

References 55 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Further studies will be necessary to follow up on these findings and titrate minimal effective concentrations of drug with activity against adult filarial worms. Interestingly, inhibition of filarial NMT proved to be very effective in our screen, confirming previous data of Galvin et al [ 27 ]. In that study, NMT was shown to be essential for viability in both B .…”

Section: Discussionsupporting

confidence: 92%

Yeast-Based High-Throughput Screens to Identify Novel Compounds Active against Brugia malayi

et al. 2016

View full text Add to dashboard Cite

BackgroundLymphatic filariasis is caused by the parasitic worms Wuchereria bancrofti, Brugia malayi or B. timori, which are transmitted via the bites from infected mosquitoes. Once in the human body, the parasites develop into adult worms in the lymphatic vessels, causing severe damage and swelling of the affected tissues. According to the World Health Organization, over 1.2 billion people in 58 countries are at risk of contracting lymphatic filariasis. Very few drugs are available to treat patients infected with these parasites, and these have low efficacy against the adult stages of the worms, which can live for 7–15 years in the human body. The requirement for annual treatment increases the risk of drug-resistant worms emerging, making it imperative to develop new drugs against these devastating diseases.Methodology/Principal FindingsWe have developed a yeast-based, high-throughput screening system whereby essential yeast genes are replaced with their filarial or human counterparts. These strains are labeled with different fluorescent proteins to allow the simultaneous monitoring of strains with parasite or human genes in competition, and hence the identification of compounds that inhibit the parasite target without affecting its human ortholog. We constructed yeast strains expressing eight different Brugia malayi drug targets (as well as seven of their human counterparts), and performed medium-throughput drug screens for compounds that specifically inhibit the parasite enzymes. Using the Malaria Box collection (400 compounds), we identified nine filarial specific inhibitors and confirmed the antifilarial activity of five of these using in vitro assays against Brugia pahangi.Conclusions/SignificanceWe were able to functionally complement yeast deletions with eight different Brugia malayi enzymes that represent potential drug targets. We demonstrated that our yeast-based screening platform is efficient in identifying compounds that can discriminate between human and filarial enzymes. Hence, we are confident that we can extend our efforts to the construction of strains with further filarial targets (in particular for those species that cannot be cultivated in the laboratory), and perform high-throughput drug screens to identify specific inhibitors of the parasite enzymes. By establishing synergistic collaborations with researchers working directly on different parasitic worms, we aim to aid antihelmintic drug development for both human and veterinary infections.

show abstract

Section: Discussionsupporting

confidence: 92%

Yeast-Based High-Throughput Screens to Identify Novel Compounds Active against Brugia malayi

et al. 2016

View full text Add to dashboard Cite

show abstract

“…The enzyme myristoyl-CoA:protein N -myristoyltransferase (NMT) has recently been validated as a promising drug target in trypanosomiasis ( Frearson et al., 2010 ) and malaria ( Wright et al., 2014 ), diseases caused by the protozoan parasites Trypanosoma brucei and species of Plasmodium , respectively. NMT has also been proposed as a potential drug target for infections caused by parasitic nematodes ( Galvin et al., 2014 ) and for the leishmaniases ( Price et al., 2003 ). NMT catalyzes a protein modification, the attachment of the C14:0 lipid myristate (Myr, tetradecanoic acid, Figure 1 A) to the N-terminal glycine residue of specific target proteins in the cell.…”

Section: Introductionmentioning

confidence: 99%

Global Analysis of Protein N-Myristoylation and Exploration of N-Myristoyltransferase as a Drug Target in the Neglected Human Pathogen Leishmania donovani

Wright

Paape

Storck

et al. 2015

Chemistry & Biology

View full text Add to dashboard Cite

SummaryN-Myristoyltransferase (NMT) modulates protein function through the attachment of the lipid myristate to the N terminus of target proteins, and is a promising drug target in eukaryotic parasites such as Leishmania donovani. Only a small number of NMT substrates have been characterized in Leishmania, and a global picture of N-myristoylation is lacking. Here, we use metabolic tagging with an alkyne-functionalized myristic acid mimetic in live parasites followed by downstream click chemistry and analysis to identify lipidated proteins in both the promastigote (extracellular) and amastigote (intracellular) life stages. Quantitative chemical proteomics is used to profile target engagement by NMT inhibitors, and to define the complement of N-myristoylated proteins. Our results provide new insight into the multiple pathways modulated by NMT and the pleiotropic effects of NMT inhibition. This work constitutes the first global experimental analysis of protein lipidation in Leishmania, and reveals the extent of NMT-related biology yet to be explored for this neglected human pathogen.

show abstract

“…N-myristoyltransferase (NMT), an enzyme which has been validated as a drug target in human cancers, and for infectious diseases caused by fungi, viruses, and protozoan parasites. Galvin et al 83 purified C. elegans and B. malayi NMTs as active recombinant proteins and carried out kinetic analyses with their essential fatty acid donor, myristoyl-CoA, and peptide substrates. The genetic and chemical studies performed in the study highlighted the importance of myristoylation in the synthesis of functional proteins in nematodes and have shown, for the first time, that NMT is required for viability in parasitic nematodes.…”

Section: Article In Pressmentioning

confidence: 99%

Targeting Human Onchocerciasis: Recent Advances Beyond Ivermectin

Sainas

Dosio

Boschi

et al. 2018

Annual Reports in Medicinal Chemistry

View full text Add to dashboard Cite

The Parasite and Its Life Cycle 1.2 Clinical Presentation 2. Drugs and Drug Combinations in Clinical or Preclinical Evaluations 2.1 Diethylcarbamazine (DEC) 2.2 Ivermectin 2.3 Benzimidazole Drugs 2.4 Doxycycline 2.5 Rifampicin 2.6 Emodepside 2.7 Combination Therapy 3. Recent Advances Beyond Ivermectin 3.1 Searching for Chemodiversity within Natural Sources 3.2 The Repositioning of Closantel: Targeting OvCHT1 Chitinase 3.3 Studies on New Therapeutic O. volvus Targets 4. Methodologies for Onchocerciasis Diagnosis 4.1 Clinical Examination 4.2 Microscopic Identification of Microfilariae in Skin Snips 4.3 Nucleic Acid-Based Molecular Assays 4.4 Diethylcarbamazine Patch Test 4.5 Antibody Diagnostic Tests 5. Conclusions References

show abstract

A Target Repurposing Approach Identifies N-myristoyltransferase as a New Candidate Drug Target in Filarial Nematodes

Cited by 21 publications

References 55 publications

Yeast-Based High-Throughput Screens to Identify Novel Compounds Active against Brugia malayi

Yeast-Based High-Throughput Screens to Identify Novel Compounds Active against Brugia malayi

Global Analysis of Protein N-Myristoylation and Exploration of N-Myristoyltransferase as a Drug Target in the Neglected Human Pathogen Leishmania donovani

Targeting Human Onchocerciasis: Recent Advances Beyond Ivermectin

Contact Info

Product

Resources

About