The heme biosynthetic pathway of the obligate Wolbachia endosymbiont of Brugia malayi as a potential anti-filarial drug target.
Filarial parasites (e.g., Brugia malayi, Onchocerca volvulus, and Wuchereria bancrofti) are causative agents of lymphatic filariasis and onchocerciasis, which are among the most disabling of neglected tropical diseases. There is an urgent need to develop macro-filaricidal drugs, as current anti-fila...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2009-07-01
|
| Series: | PLoS Neglected Tropical Diseases |
| Online Access: | http://europepmc.org/articles/PMC2703803?pdf=render |
| _version_ | 1811196196117544960 |
|---|---|
| author | Bo Wu Jacopo Novelli Jeremy Foster Romualdas Vaisvila Leslie Conway Jessica Ingram Mehul Ganatra Anita U Rao Iqbal Hamza Barton Slatko |
| author_facet | Bo Wu Jacopo Novelli Jeremy Foster Romualdas Vaisvila Leslie Conway Jessica Ingram Mehul Ganatra Anita U Rao Iqbal Hamza Barton Slatko |
| author_sort | Bo Wu |
| collection | DOAJ |
| description | Filarial parasites (e.g., Brugia malayi, Onchocerca volvulus, and Wuchereria bancrofti) are causative agents of lymphatic filariasis and onchocerciasis, which are among the most disabling of neglected tropical diseases. There is an urgent need to develop macro-filaricidal drugs, as current anti-filarial chemotherapy (e.g., diethylcarbamazine [DEC], ivermectin and albendazole) can interrupt transmission predominantly by killing microfilariae (mf) larvae, but is less effective on adult worms, which can live for decades in the human host. All medically relevant human filarial parasites appear to contain an obligate endosymbiotic bacterium, Wolbachia. This alpha-proteobacterial mutualist has been recognized as a potential target for filarial nematode life cycle intervention, as antibiotic treatments of filarial worms harboring Wolbachia result in the loss of worm fertility and viability upon antibiotic treatments both in vitro and in vivo. Human trials have confirmed this approach, although the length of treatments, high doses required and medical counter-indications for young children and pregnant women warrant the identification of additional anti-Wolbachia drugs.Genome sequence analysis indicated that enzymes involved in heme biosynthesis might constitute a potential anti-Wolbachia target set. We tested different heme biosynthetic pathway inhibitors in ex vivo B. malayi viability assays and report a specific effect of N-methyl mesoporphyrin (NMMP), which targets ferrochelatase (FC, the last step). Our phylogenetic analysis indicates evolutionarily significant divergence between Wolbachia heme genes and their human homologues. We therefore undertook the cloning, overexpression and analysis of several enzymes of this pathway alongside their human homologues, and prepared proteins for drug targeting. In vitro enzyme assays revealed a approximately 600-fold difference in drug sensitivities to succinyl acetone (SA) between Wolbachia and human 5'-aminolevulinic acid dehydratase (ALAD, the second step). Similarly, Escherichia coli hemH (FC) deficient strains transformed with human and Wolbachia FC homologues showed significantly different sensitivities to NMMP. This approach enables functional complementation in E. coli heme deficient mutants as an alternative E. coli-based method for drug screening.Our studies indicate that the heme biosynthetic genes in the Wolbachia of B. malayi (wBm) might be essential for the filarial host survival. In addition, the results suggest they are likely candidate drug targets based upon significant differences in phylogenetic distance, biochemical properties and sensitivities to heme biosynthesis inhibitors, as compared to their human homologues. |
| first_indexed | 2024-04-12T00:54:33Z |
| format | Article |
| id | doaj.art-5a46d746c1184bb1ae13e0c5dd598142 |
| institution | Directory Open Access Journal |
| issn | 1935-2727 1935-2735 |
| language | English |
| last_indexed | 2024-04-12T00:54:33Z |
| publishDate | 2009-07-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Neglected Tropical Diseases |
| spelling | doaj.art-5a46d746c1184bb1ae13e0c5dd5981422022-12-22T03:54:38ZengPublic Library of Science (PLoS)PLoS Neglected Tropical Diseases1935-27271935-27352009-07-0137e47510.1371/journal.pntd.0000475The heme biosynthetic pathway of the obligate Wolbachia endosymbiont of Brugia malayi as a potential anti-filarial drug target.Bo WuJacopo NovelliJeremy FosterRomualdas VaisvilaLeslie ConwayJessica IngramMehul GanatraAnita U RaoIqbal HamzaBarton SlatkoFilarial parasites (e.g., Brugia malayi, Onchocerca volvulus, and Wuchereria bancrofti) are causative agents of lymphatic filariasis and onchocerciasis, which are among the most disabling of neglected tropical diseases. There is an urgent need to develop macro-filaricidal drugs, as current anti-filarial chemotherapy (e.g., diethylcarbamazine [DEC], ivermectin and albendazole) can interrupt transmission predominantly by killing microfilariae (mf) larvae, but is less effective on adult worms, which can live for decades in the human host. All medically relevant human filarial parasites appear to contain an obligate endosymbiotic bacterium, Wolbachia. This alpha-proteobacterial mutualist has been recognized as a potential target for filarial nematode life cycle intervention, as antibiotic treatments of filarial worms harboring Wolbachia result in the loss of worm fertility and viability upon antibiotic treatments both in vitro and in vivo. Human trials have confirmed this approach, although the length of treatments, high doses required and medical counter-indications for young children and pregnant women warrant the identification of additional anti-Wolbachia drugs.Genome sequence analysis indicated that enzymes involved in heme biosynthesis might constitute a potential anti-Wolbachia target set. We tested different heme biosynthetic pathway inhibitors in ex vivo B. malayi viability assays and report a specific effect of N-methyl mesoporphyrin (NMMP), which targets ferrochelatase (FC, the last step). Our phylogenetic analysis indicates evolutionarily significant divergence between Wolbachia heme genes and their human homologues. We therefore undertook the cloning, overexpression and analysis of several enzymes of this pathway alongside their human homologues, and prepared proteins for drug targeting. In vitro enzyme assays revealed a approximately 600-fold difference in drug sensitivities to succinyl acetone (SA) between Wolbachia and human 5'-aminolevulinic acid dehydratase (ALAD, the second step). Similarly, Escherichia coli hemH (FC) deficient strains transformed with human and Wolbachia FC homologues showed significantly different sensitivities to NMMP. This approach enables functional complementation in E. coli heme deficient mutants as an alternative E. coli-based method for drug screening.Our studies indicate that the heme biosynthetic genes in the Wolbachia of B. malayi (wBm) might be essential for the filarial host survival. In addition, the results suggest they are likely candidate drug targets based upon significant differences in phylogenetic distance, biochemical properties and sensitivities to heme biosynthesis inhibitors, as compared to their human homologues.http://europepmc.org/articles/PMC2703803?pdf=render |
| spellingShingle | Bo Wu Jacopo Novelli Jeremy Foster Romualdas Vaisvila Leslie Conway Jessica Ingram Mehul Ganatra Anita U Rao Iqbal Hamza Barton Slatko The heme biosynthetic pathway of the obligate Wolbachia endosymbiont of Brugia malayi as a potential anti-filarial drug target. PLoS Neglected Tropical Diseases |
| title | The heme biosynthetic pathway of the obligate Wolbachia endosymbiont of Brugia malayi as a potential anti-filarial drug target. |
| title_full | The heme biosynthetic pathway of the obligate Wolbachia endosymbiont of Brugia malayi as a potential anti-filarial drug target. |
| title_fullStr | The heme biosynthetic pathway of the obligate Wolbachia endosymbiont of Brugia malayi as a potential anti-filarial drug target. |
| title_full_unstemmed | The heme biosynthetic pathway of the obligate Wolbachia endosymbiont of Brugia malayi as a potential anti-filarial drug target. |
| title_short | The heme biosynthetic pathway of the obligate Wolbachia endosymbiont of Brugia malayi as a potential anti-filarial drug target. |
| title_sort | heme biosynthetic pathway of the obligate wolbachia endosymbiont of brugia malayi as a potential anti filarial drug target |
| url | http://europepmc.org/articles/PMC2703803?pdf=render |
| work_keys_str_mv | AT bowu thehemebiosyntheticpathwayoftheobligatewolbachiaendosymbiontofbrugiamalayiasapotentialantifilarialdrugtarget AT jacoponovelli thehemebiosyntheticpathwayoftheobligatewolbachiaendosymbiontofbrugiamalayiasapotentialantifilarialdrugtarget AT jeremyfoster thehemebiosyntheticpathwayoftheobligatewolbachiaendosymbiontofbrugiamalayiasapotentialantifilarialdrugtarget AT romualdasvaisvila thehemebiosyntheticpathwayoftheobligatewolbachiaendosymbiontofbrugiamalayiasapotentialantifilarialdrugtarget AT leslieconway thehemebiosyntheticpathwayoftheobligatewolbachiaendosymbiontofbrugiamalayiasapotentialantifilarialdrugtarget AT jessicaingram thehemebiosyntheticpathwayoftheobligatewolbachiaendosymbiontofbrugiamalayiasapotentialantifilarialdrugtarget AT mehulganatra thehemebiosyntheticpathwayoftheobligatewolbachiaendosymbiontofbrugiamalayiasapotentialantifilarialdrugtarget AT anitaurao thehemebiosyntheticpathwayoftheobligatewolbachiaendosymbiontofbrugiamalayiasapotentialantifilarialdrugtarget AT iqbalhamza thehemebiosyntheticpathwayoftheobligatewolbachiaendosymbiontofbrugiamalayiasapotentialantifilarialdrugtarget AT bartonslatko thehemebiosyntheticpathwayoftheobligatewolbachiaendosymbiontofbrugiamalayiasapotentialantifilarialdrugtarget AT bowu hemebiosyntheticpathwayoftheobligatewolbachiaendosymbiontofbrugiamalayiasapotentialantifilarialdrugtarget AT jacoponovelli hemebiosyntheticpathwayoftheobligatewolbachiaendosymbiontofbrugiamalayiasapotentialantifilarialdrugtarget AT jeremyfoster hemebiosyntheticpathwayoftheobligatewolbachiaendosymbiontofbrugiamalayiasapotentialantifilarialdrugtarget AT romualdasvaisvila hemebiosyntheticpathwayoftheobligatewolbachiaendosymbiontofbrugiamalayiasapotentialantifilarialdrugtarget AT leslieconway hemebiosyntheticpathwayoftheobligatewolbachiaendosymbiontofbrugiamalayiasapotentialantifilarialdrugtarget AT jessicaingram hemebiosyntheticpathwayoftheobligatewolbachiaendosymbiontofbrugiamalayiasapotentialantifilarialdrugtarget AT mehulganatra hemebiosyntheticpathwayoftheobligatewolbachiaendosymbiontofbrugiamalayiasapotentialantifilarialdrugtarget AT anitaurao hemebiosyntheticpathwayoftheobligatewolbachiaendosymbiontofbrugiamalayiasapotentialantifilarialdrugtarget AT iqbalhamza hemebiosyntheticpathwayoftheobligatewolbachiaendosymbiontofbrugiamalayiasapotentialantifilarialdrugtarget AT bartonslatko hemebiosyntheticpathwayoftheobligatewolbachiaendosymbiontofbrugiamalayiasapotentialantifilarialdrugtarget |