Enolase inhibitors as therapeutic leads for Naegleria fowleri infection.
Infections with the pathogenic free-living amoebae Naegleria fowleri can lead to life-threatening illnesses including catastrophic primary amoebic meningoencephalitis (PAM). Efficacious treatment options for these infections are lacking and the mortality rate remains >95% in the US. Glycolysis is...
Saved in:
| Main Authors: | , , , , , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2024-08-01
|
| Series: | PLoS Pathogens |
| Online Access: | https://doi.org/10.1371/journal.ppat.1012412 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1841533266072436736 |
|---|---|
| author | Jillian E Milanes Victoria C Yan Cong-Dat Pham Florian Muller Samuel Kwain Kerrick C Rees Brian N Dominy Daniel C Whitehead Steven W Millward Madison Bolejack Roger Shek Logan Tillery Isabelle Q Phan Bart Staker E Ashley Moseman Xiang Zhang Xipeng Ma Audriy Jebet Xinmin Yin James C Morris |
| author_facet | Jillian E Milanes Victoria C Yan Cong-Dat Pham Florian Muller Samuel Kwain Kerrick C Rees Brian N Dominy Daniel C Whitehead Steven W Millward Madison Bolejack Roger Shek Logan Tillery Isabelle Q Phan Bart Staker E Ashley Moseman Xiang Zhang Xipeng Ma Audriy Jebet Xinmin Yin James C Morris |
| author_sort | Jillian E Milanes |
| collection | DOAJ |
| description | Infections with the pathogenic free-living amoebae Naegleria fowleri can lead to life-threatening illnesses including catastrophic primary amoebic meningoencephalitis (PAM). Efficacious treatment options for these infections are lacking and the mortality rate remains >95% in the US. Glycolysis is very important for the infectious trophozoite lifecycle stage and inhibitors of glucose metabolism have been found to be toxic to the pathogen. Recently, human enolase 2 (ENO2) phosphonate inhibitors have been developed as lead agents to treat glioblastoma multiforme (GBM). These compounds, which cure GBM in a rodent model, are well-tolerated in mammals because enolase 1 (ENO1) is the predominant isoform used systemically. Here, we describe findings that demonstrate these agents are potent inhibitors of N. fowleri ENO (NfENO) and are lethal to amoebae. In particular, (1-hydroxy-2-oxopiperidin-3-yl) phosphonic acid (HEX) was a potent enzyme inhibitor (IC50 = 0.14 ± 0.04 μM) that was toxic to trophozoites (EC50 = 0.21 ± 0.02 μM) while the reported CC50 was >300 μM. Molecular docking simulation revealed that HEX binds strongly to the active site of NfENO with a binding affinity of -8.6 kcal/mol. Metabolomic studies of parasites treated with HEX revealed a 4.5 to 78-fold accumulation of glycolytic intermediates upstream of NfENO. Last, nasal instillation of HEX increased longevity of amoebae-infected rodents. Two days after infection, animals were treated for 10 days with 3 mg/kg HEX, followed by one week of observation. At the end of the one-week observation, eight of 12 HEX-treated animals remained alive (resulting in an indeterminable median survival time) while one of 12 vehicle-treated rodents remained, yielding a median survival time of 10.9 days. However, intranasal HEX delivery was not curative as brains of six of the eight survivors were positive for amoebae. These findings suggest that HEX requires further evaluation to develop as a lead for treatment of PAM. |
| format | Article |
| id | doaj-art-cb6c340b2bff4f6ca9f6beb1b6aa1504 |
| institution | Kabale University |
| issn | 1553-7366 1553-7374 |
| language | English |
| publishDate | 2024-08-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Pathogens |
| spelling | doaj-art-cb6c340b2bff4f6ca9f6beb1b6aa15042025-01-17T05:31:04ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742024-08-01208e101241210.1371/journal.ppat.1012412Enolase inhibitors as therapeutic leads for Naegleria fowleri infection.Jillian E MilanesVictoria C YanCong-Dat PhamFlorian MullerSamuel KwainKerrick C ReesBrian N DominyDaniel C WhiteheadSteven W MillwardMadison BolejackRoger ShekLogan TilleryIsabelle Q PhanBart StakerE Ashley MosemanXiang ZhangXipeng MaAudriy JebetXinmin YinJames C MorrisInfections with the pathogenic free-living amoebae Naegleria fowleri can lead to life-threatening illnesses including catastrophic primary amoebic meningoencephalitis (PAM). Efficacious treatment options for these infections are lacking and the mortality rate remains >95% in the US. Glycolysis is very important for the infectious trophozoite lifecycle stage and inhibitors of glucose metabolism have been found to be toxic to the pathogen. Recently, human enolase 2 (ENO2) phosphonate inhibitors have been developed as lead agents to treat glioblastoma multiforme (GBM). These compounds, which cure GBM in a rodent model, are well-tolerated in mammals because enolase 1 (ENO1) is the predominant isoform used systemically. Here, we describe findings that demonstrate these agents are potent inhibitors of N. fowleri ENO (NfENO) and are lethal to amoebae. In particular, (1-hydroxy-2-oxopiperidin-3-yl) phosphonic acid (HEX) was a potent enzyme inhibitor (IC50 = 0.14 ± 0.04 μM) that was toxic to trophozoites (EC50 = 0.21 ± 0.02 μM) while the reported CC50 was >300 μM. Molecular docking simulation revealed that HEX binds strongly to the active site of NfENO with a binding affinity of -8.6 kcal/mol. Metabolomic studies of parasites treated with HEX revealed a 4.5 to 78-fold accumulation of glycolytic intermediates upstream of NfENO. Last, nasal instillation of HEX increased longevity of amoebae-infected rodents. Two days after infection, animals were treated for 10 days with 3 mg/kg HEX, followed by one week of observation. At the end of the one-week observation, eight of 12 HEX-treated animals remained alive (resulting in an indeterminable median survival time) while one of 12 vehicle-treated rodents remained, yielding a median survival time of 10.9 days. However, intranasal HEX delivery was not curative as brains of six of the eight survivors were positive for amoebae. These findings suggest that HEX requires further evaluation to develop as a lead for treatment of PAM.https://doi.org/10.1371/journal.ppat.1012412 |
| spellingShingle | Jillian E Milanes Victoria C Yan Cong-Dat Pham Florian Muller Samuel Kwain Kerrick C Rees Brian N Dominy Daniel C Whitehead Steven W Millward Madison Bolejack Roger Shek Logan Tillery Isabelle Q Phan Bart Staker E Ashley Moseman Xiang Zhang Xipeng Ma Audriy Jebet Xinmin Yin James C Morris Enolase inhibitors as therapeutic leads for Naegleria fowleri infection. PLoS Pathogens |
| title | Enolase inhibitors as therapeutic leads for Naegleria fowleri infection. |
| title_full | Enolase inhibitors as therapeutic leads for Naegleria fowleri infection. |
| title_fullStr | Enolase inhibitors as therapeutic leads for Naegleria fowleri infection. |
| title_full_unstemmed | Enolase inhibitors as therapeutic leads for Naegleria fowleri infection. |
| title_short | Enolase inhibitors as therapeutic leads for Naegleria fowleri infection. |
| title_sort | enolase inhibitors as therapeutic leads for naegleria fowleri infection |
| url | https://doi.org/10.1371/journal.ppat.1012412 |
| work_keys_str_mv | AT jillianemilanes enolaseinhibitorsastherapeuticleadsfornaegleriafowleriinfection AT victoriacyan enolaseinhibitorsastherapeuticleadsfornaegleriafowleriinfection AT congdatpham enolaseinhibitorsastherapeuticleadsfornaegleriafowleriinfection AT florianmuller enolaseinhibitorsastherapeuticleadsfornaegleriafowleriinfection AT samuelkwain enolaseinhibitorsastherapeuticleadsfornaegleriafowleriinfection AT kerrickcrees enolaseinhibitorsastherapeuticleadsfornaegleriafowleriinfection AT brianndominy enolaseinhibitorsastherapeuticleadsfornaegleriafowleriinfection AT danielcwhitehead enolaseinhibitorsastherapeuticleadsfornaegleriafowleriinfection AT stevenwmillward enolaseinhibitorsastherapeuticleadsfornaegleriafowleriinfection AT madisonbolejack enolaseinhibitorsastherapeuticleadsfornaegleriafowleriinfection AT rogershek enolaseinhibitorsastherapeuticleadsfornaegleriafowleriinfection AT logantillery enolaseinhibitorsastherapeuticleadsfornaegleriafowleriinfection AT isabelleqphan enolaseinhibitorsastherapeuticleadsfornaegleriafowleriinfection AT bartstaker enolaseinhibitorsastherapeuticleadsfornaegleriafowleriinfection AT eashleymoseman enolaseinhibitorsastherapeuticleadsfornaegleriafowleriinfection AT xiangzhang enolaseinhibitorsastherapeuticleadsfornaegleriafowleriinfection AT xipengma enolaseinhibitorsastherapeuticleadsfornaegleriafowleriinfection AT audriyjebet enolaseinhibitorsastherapeuticleadsfornaegleriafowleriinfection AT xinminyin enolaseinhibitorsastherapeuticleadsfornaegleriafowleriinfection AT jamescmorris enolaseinhibitorsastherapeuticleadsfornaegleriafowleriinfection |