bims-toxgon 2023-06-04 papers

bims-toxgon

Biomed News

on Toxoplasma gondii metabolism

Issue of 2023–06–04
ten papers selected by
Lakesh Kumar, BITS Pilani

Overexpression of Eimeria tenella Rhoptry Kinase 2 Induces Early Production of Schizonts.
Expression of cytokines and co-stimulatory molecules in the Toxoplasma gondii-infected dendritic cells of C57BL/6 and BALB/c mice.
An essential role for an Fe-S cluster protein in the cytochrome c oxidase complex of Toxoplasma parasites.
Anti-Toxoplasma gondii activity of Trametes versicolor (Turkey tail) mushroom extract.
Host-pathogen interactions mediated by extracellular vesicles in Toxoplasma gondii infection during pregnancy.
Both maternal IFNγ exposure and acute prenatal infection with Toxoplasma gondii activate fetal hematopoietic stem cells.
Toxoplasma and Toxocara seropositivity in juvenile idiopathic arthritis and its relation to disease activity and type of therapies.
Toxoplasma gondii attenuates the ethidium bromide induced demyelination lesions in multiple sclerosis model rats.
ULK1-mediated metabolic reprogramming regulates Vps34 lipid kinase activity by its lactylation.
Post-translational Modifications by Acyl Groups Regulate FEN1's Activities and Play Essential Roles in Cell Proliferation and DNA Repair.

Microbiol Spectr. 2023 Jun 01. e0013723

Overexpression of Eimeria tenella Rhoptry Kinase 2 Induces Early Production of Schizonts.

Adeline Ribeiro E Silva, Mamadou Amadou Diallo, Alix Sausset, Thomas Robert, Stéphane Bach, Françoise I Bussière, Fabrice Laurent, Sonia Lacroix-Lamandé, Anne Silvestre.

  Eimeria tenella is an obligate intracellular parasite responsible for avian coccidiosis. Like other apicomplexan parasites, such as Toxoplasma gondii, cell invasion and intracellular development rely on apical organelle content discharge, named micronemes and rhoptries. Some rhoptry (ROP) kinases (ROPK) are key virulence factors in T. gondii. To date, among the 28 ropk genes carried by E. tenella, only two to four were confirmed by proteomic analysis or immunostaining to be expressed at the sporozoite stage. We have previously shown that EtROP1 is implicated in the inhibition of host cell apoptosis by interacting with the cellular p53. This work functionally described the second ROP kinase expressed at the sporozoite stage in E. tenella. EtROP2 is an active kinase that phosphorylates cell substrates of approximately 50 kDa. Its overexpression leads to the shortening of the prepatent period and to the early development of first-generation schizonts. Conduction of RNA sequencing analysis and reverse transcriptase quantitative PCR (RT-qPCR) on the host cell allowed us to identify the mitogen-activated protein kinase (MAPK) pathway and the transcription factor cFos to be upregulated by EtROP2. We also showed by immunofluorescence assay that the active kinase EtROP2 is implicated in the p38 MAPK pathway activation. We established here that EtROP2 activates the p38 MAPK pathway through a direct or indirect phosphorylation, leading to the overexpression of the master transcription factor cFos known to be implicated in E. tenella development. IMPORTANCE Rhoptries are specialized secretory organelles found in zoite stages of apicomplexan parasites. In addition to well-conserved rhoptry neck proteins, their protein consists mostly of kinase proteins, highly divergent from eukaryotic kinases. Some of those kinases are described as major virulence factors in Toxoplasma gondii, secreted into the host cell to hijack signaling pathways. Most of those kinases remain to be characterized in Eimeria tenella. Deciphering their cellular function is a prerequisite to supporting their relevance as a druggable target in development of new means of Eimeria tenella control. Secreted divergent kinases that interact with host cell partners to modulate pathways are good candidates, as they coevolve with their host targets to ensure their function within the host and are less prone to mutations that would lead to drug resistance. The absence of any orthologous kinase in host cells makes these parasite kinases a promising drug target candidate.

Keywords:  Apicomplexa; Eimeria; ROPK; kinase; p38 MAPK; rhoptry

DOI:  https://doi.org/10.1128/spectrum.00137-23
Parasites Hosts Dis. 2023 May;61(2): 138-146

Expression of cytokines and co-stimulatory molecules in the Toxoplasma gondii-infected dendritic cells of C57BL/6 and BALB/c mice.

Jae-Hyung Lee, Jae-Min Yuk, Guang-Ho Cha, Young-Ha Lee.

  Toxoplasma gondii is an intracellular protozoan parasite which can infect most warm-blooded animals and humans. Among the different mouse models, C57BL/6 mice are more susceptible to T. gondii infection compared to BALB/c mice, and this increased susceptibility has been attributed to various factors, including T-cell responses. Dendritic cells (DCs) are the most prominent type of antigen-presenting cells and regulate the host immune response, including the response of T-cells. However, differences in the DC responses of these mouse strains to T. gondii infection have yet to be characterized. In this study, we cultured bone marrow-derived DCs (BMDCs) from BALB/c and C57BL/6 mice. These cells were infected with T. gondii. The activation of the BMDCs was assessed based on the expression of cell surface markers and cytokines. In the BMDCs of both mouse strains, we detected significant increases in the expression of cell surface T-cell co-stimulatory molecules (major histocompatibility complex (MHC) II, CD40, CD80, and CD86) and cytokines (tumor necrosis factor (TNF)-α, interferon (IFN)-γ, interleukin (IL)-12p40, IL-1β, and IL-10) from 3 h post-T. gondii infection. The expression of MHC II, CD40, CD80, CD86, IFN-γ, IL-12p40, and IL-1β was significantly higher in the T. gondii-infected BMDCs obtained from the C57BL/6 mice than in those from the BALB/c mice. These findings indicate that differences in the activation status of the BMDCs in the BALB/c and C57BL/6 mice may account for their differential susceptibility to T. gondii.

Keywords:  Toxoplasma gondii; co-stimulatory molecule; cytokine; dendritic cell; mouse

DOI:  https://doi.org/10.3347/PHD.22150
PLoS Pathog. 2023 Jun 01. 19(6): e1011430

An essential role for an Fe-S cluster protein in the cytochrome c oxidase complex of Toxoplasma parasites.

Rachel A Leonard, Yuan Tian, Feng Tan, Giel G van Dooren, Jenni A Hayward.

The mitochondrial electron transport chain (ETC) of apicomplexan parasites differs considerably from the ETC of the animals that these parasites infect, and is the target of numerous anti-parasitic drugs. The cytochrome c oxidase complex (Complex IV) of the apicomplexan Toxoplasma gondii ETC is more than twice the mass and contains subunits not found in human Complex IV, including a 13 kDa protein termed TgApiCox13. TgApiCox13 is homologous to a human iron-sulfur (Fe-S) cluster-containing protein called the mitochondrial inner NEET protein (HsMiNT) which is not a component of Complex IV in humans. Here, we establish that TgApiCox13 is a critical component of Complex IV in T. gondii, required for complex activity and stability. Furthermore, we demonstrate that TgApiCox13, like its human homolog, binds two Fe-S clusters. We show that the Fe-S clusters of TgApiCox13 are critical for ETC function, having an essential role in mediating Complex IV integrity. Our study provides the first functional characterisation of an Fe-S protein in Complex IV.

DOI: https://doi.org/10.1371/journal.ppat.1011430
Sci Rep. 2023 05 29. 13(1): 8667

Anti-Toxoplasma gondii activity of Trametes versicolor (Turkey tail) mushroom extract.

Homa Nath Sharma, Jonathan Catrett, Ogechi Destiny Nwokeocha, Melissa Boersma, Michael E Miller, Audrey Napier, Boakai K Robertson, Daniel A Abugri.

Toxoplasma gondii (T. gondii) infection continues to rise globally in humans and animals with high socioeconomic and public health challenges. Current medications used against T. gondii infection are limited in efficacy, safety, and affordability. This research was conducted to assess the higher fungi extract effect on T. gondii tachyzoites growth in vitro and possibly decipher its mechanism of action. Furthermore, we evaluated the extract's effect on human foreskin fibroblast viability. The methanol extracts of Turkey tail (TT) mushroom was tested against T. gondii tachyzoites growth using an RH-RFP type I strain that expresses red fluorescent protein throughout culture in a dose-dependent manner using a fluorescent plate reader. Similarly, we tested the effect of the extract on host cell viability. We observed that TT extract inhibited tachyzoites growth with a 50% minimum inhibitory concentration (IC50s), IC50 = 5.98 ± 1.22 µg/mL, and 50% cytotoxic concentration (CC50s), CC50 ≥ 100 µg/mL. It was discovered that TT extract induced strong mitochondria superoxide and reactive oxygen species production and disrupted mitochondria membrane potential in T. gondii tachyzoites. Additionally, scanning electron microscopy depicted that TT extract and pyrimethamine (PY) caused a morphological deformation of tachyzoites in vitro. In conclusion, TT methanol extract made up of phytosterols, bioactive sphingolipids, peptides, phenolic acids, and lactones could be a promising source of new compounds for the future development of anti-Toxoplasma gondii drugs. Extracts were non-cytotoxic, even at higher concentrations.

DOI: https://doi.org/10.1038/s41598-023-35676-6
J Reprod Immunol. 2023 May 18. pii: S0165-0378(23)00163-8. [Epub ahead of print]158 103957

Host-pathogen interactions mediated by extracellular vesicles in Toxoplasma gondii infection during pregnancy.

Fernando Gómez-Chávez, José M Murrieta-Coxca, Heriberto Caballero-Ortega, Diana M Morales-Prieto, Udo R Markert.

  Molecular communication between a pathogen and its host is crucial for a successful interplay. Extracellular vesicles (EVs) act as mediators for the delivery of molecular signals among pathogens or between pathogens and the host. Toxoplasma gondii (T. gondii), an intracellular parasite with a worldwide presence, produces EVs itself, or induces the secretion of EVs from infected host cells potentially having capacities to modulate the host immune response. T. gondii infection is particularly important during pregnancy. Depending on the gestational age at the time of infection, the parasite can be transmitted through the placenta to the fetus, causing clinical complications such as jaundice, hepatosplenomegaly, chorioretinitis, cranioencephalic abnormalities, or even death. T. gondii infection is related to a pro-inflammatory immune response in both mother and fetus, which may enhance parasite transmission, but the implication of EV signaling in this process remains unclear. In this review, we summarize the current knowledge on EV release from T. gondii and its human host cells in regard to the immunological consequences and the passage through the placenta.

Keywords:  Extracellular vesicles; Immune cells; Placenta; Pregnancy; Toxoplasma

DOI:  https://doi.org/10.1016/j.jri.2023.103957
EMBO J. 2023 Jun 01. e112693

Both maternal IFNγ exposure and acute prenatal infection with Toxoplasma gondii activate fetal hematopoietic stem cells.

Diego A López, Kelly S Otsuka, April C Apostol, Jasmine Posada, Juan C Sánchez-Arcila, Kirk Dc Jensen, Anna E Beaudin.

  Infection directly influences adult hematopoietic stem cell (HSC) function and differentiation, but the fetal hematopoietic response to infection during pregnancy is not well-studied. Here, we investigated the fetal hematopoietic response to maternal infection with Toxoplasma gondii (T. gondii), an intracellular parasite that elicits Type II IFNγ-mediated maternal immunity. While it is known that maternal infection without direct pathogen transmission can affect fetal immune development, the effects of maternal IFNγ on developing HSCs and the signals that mediate these interactions have not been investigated. Our investigation reveals that the fetal HSCs respond to T. gondii infection with virulence-dependent changes in proliferation, self-renewal potential, and lineage output. Furthermore, maternal IFNγ crosses the fetal-maternal interface, where it is perceived by fetal HSCs. By comparing the effects of maternal IFNγ injection with maternal T. gondii infection, we reveal that the effects of IFNγ treatment mimic some aspects of the fetal HSC response to infection. Moreover, our findings illuminate that the fetal HSC response to prenatal infection is distinct from the adult HSC response to IFNγ-induced inflammation. Altogether, our data disentangle the role of infection-induced inflammatory cytokines in driving the expansion of downstream hematopoietic progenitors.

Keywords:  Toxoplasma gondii; congenital infection; hematopoiesis; hematopoietic stem cell; inflammation

DOI:  https://doi.org/10.15252/embj.2022112693
Food Waterborne Parasitol. 2023 Jun;31 e00195

Toxoplasma and Toxocara seropositivity in juvenile idiopathic arthritis and its relation to disease activity and type of therapies.

Doaa A Salem, Ahmed Hassan Al-Ghamdi, Jameel Mohamed Alghamdi, Amira Ismail, Bakheet A Alghamdi, Eman Abdelrazek.

  Juvenile idiopathic arthritis (JIA) is the most frequently encountered autoimmune rheumatic disease in children. To our knowledge, this is the first study aimed to estimate the frequency of Toxoplasma gondii (T. gondii) and Toxocara seropositivity in JIA and assess its relation to the disease activity, IL-10 levels, and type of the received therapies. This study was conducted on 43 JIA patients and 50 cases as a control group. All participants were evaluated by disease activity score (JADAS-27), and the presence of specific IgG and IgM antibodies against T. gondii and IgG against Toxocara species using an enzyme-linked immunosorbent assay. IL-10 serum levels were measured using an ELISA kit. The results show that JIA patients have significantly higher seropositivity for anti-T. gondii IgG compared to control subjects (p = 0.02) and a non-significant difference for Toxocara seropositivity (p = 0.41). All participants were negative for IgM anti-Toxoplasma gondii. Demographic parameters did not significantly affect these seroprevalence frequencies (p > 0.05). IL-10 was significantly higher among JIA patients compared to controls (p = 0.007) and seropositive anti-T. gondii JIA exhibited significantly higher IL-10 levels compared to seronegative ones (p = 0.03). Seropositive anti-T. gondii IgG JIA patients had a significantly higher disease activity score (JADAS-27) than seronegative anti-T. gondii IgG cases (p = 0.02). There was a significant positive correlation between anti-T. gondii IgG and JADAS-27 score (p = 0.009). A significant association was detected between T. gondii infection and DMARDs including the biological therapies (p < 0.05). Overall, this study supports a possible association between T. gondii infection and JIA, IL-10, disease activity score, and DMARDs therapies. It is possible that IL-10 plays a role in the development of JIA and contributes to persistent asymptomatic infection with T. gondii in JIA patients. As a result, a recommendation for screening tests for T. gondii infection among JIA patients is crucial before and during commencing DMARDs therapies and closely monitoring early signs of infection.

Keywords:  Autoimmunity; Biological therapies; Interleukin-10; Juvenile idiopathic arthritis; Seroprevalence; Toxocara; Toxoplasma gondii

DOI:  https://doi.org/10.1016/j.fawpar.2023.e00195
Int Immunopharmacol. 2023 May 25. pii: S1567-5769(23)00702-6. [Epub ahead of print]120 110379

Toxoplasma gondii attenuates the ethidium bromide induced demyelination lesions in multiple sclerosis model rats.

Seyedmousa Motavallihaghi, Mojgan Ghaemipanaeian, Sara Soleimani Asl, Faeze Foroughi-Parvar, Amir Hossein Maghsood.

  Multiple sclerosis (MS) is an autoimmune neurodegenerative disease. Since the modulation of the immune system by parasites has been proven, and there have been reports of a reduction in the clinical symptoms of MS in people with toxoplasmosis, this study aimed to investigate the effect of toxoplasmosis on MS in an animal model. MS model was induced by the ethidium bromide injection in the areas specified in the Rat's brain in the stereotaxic device and Toxoplasma gondii RH strain injection of the rat's peritoneal for creation of toxoplasmosis. The effect of acute and chronic toxoplasmosis on the MS model was evaluated by examining the development of clinical symptoms of MS, body weight, changes in the levels of inflammatory cytokines, inflammatory cell infiltration, cell density, and spongy tissue in the brain. The body weight in the acute toxoplasmosis with MS was the same as the MS group, and a significant decrease was observed, but no weight loss was observed in the chronic toxoplasmosis with MS. In the chronic toxoplasmosis, the progress of clinical signs such as Immobility of limbs, including tail, hands, and feet, was observed less compared to other groups. The histology results in the group of chronic toxoplasmosis showed high cell density and inhibition of spongy tissue formation, and the infiltration of inflammatory cells in this group was less. TNF-α and INF-γ decreased in MS with chronic toxoplasmosis compared to the MS group. Our findings showed that chronic toxoplasmosis with inhibition of spongy tissue formation and prevention of cell infiltration and. As a result, the reduction of inflammatory cytokines could reduce clinical symptoms in MS in the animal model.

Keywords:  Demyelination; MS; Multiple sclerosis; Toxoplasma gondii

DOI:  https://doi.org/10.1016/j.intimp.2023.110379
Sci Adv. 2023 Jun 02. 9(22): eadg4993

ULK1-mediated metabolic reprogramming regulates Vps34 lipid kinase activity by its lactylation.

Mengshu Jia, Xiao Yue, Weixia Sun, Qianjun Zhou, Cheng Chang, Weihua Gong, Jian Feng, Xie Li, Ruonan Zhan, Kemin Mo, Lijuan Zhang, Yajie Qian, Yuying Sun, Aoxue Wang, Yejun Zou, Weicai Chen, Yan Li, Li Huang, Yi Yang, Yuzheng Zhao, Xiawei Cheng.

Autophagy and glycolysis are highly conserved biological processes involved in both physiological and pathological cellular programs, but the interplay between these processes is poorly understood. Here, we show that the glycolytic enzyme lactate dehydrogenase A (LDHA) is activated upon UNC-51-like kinase 1 (ULK1) activation under nutrient deprivation. Specifically, ULK1 directly interacts with LDHA, phosphorylates serine-196 when nutrients are scarce and promotes lactate production. Lactate connects autophagy and glycolysis through Vps34 lactylation (at lysine-356 and lysine-781), which is mediated by the acyltransferase KAT5/TIP60. Vps34 lactylation enhances the association of Vps34 with Beclin1, Atg14L, and UVRAG, and then increases Vps34 lipid kinase activity. Vps34 lactylation promotes autophagic flux and endolysosomal trafficking. Vps34 lactylation in skeletal muscle during intense exercise maintains muscle cell homeostasis and correlates with cancer progress by inducing cell autophagy. Together, our findings describe autophagy regulation mechanism and then integrate cell autophagy and glycolysis.

DOI: https://doi.org/10.1126/sciadv.adg4993
Protein Pept Lett. 2023 May 29.

Post-translational Modifications by Acyl Groups Regulate FEN1's Activities and Play Essential Roles in Cell Proliferation and DNA Repair.

Yue Xiao, Mingyu Yin, Yiyi Wang, Rongyi Shi, Shuyu Mao, Yuejin Hua, Hong Xu.

   BACKGROUND: Flap endonuclease 1 (FEN1), well known for its structural-specific nuclease, possessing 5'-flap endonuclease and 5'-3' exonuclease ac-tivities, is mainly involved in DNA replication and repair. Protein lysine acetylation is an important posttranslational modification that could regulate numerous proteins' activity, subcellular localization, protein-protein interac-tion etc., and influences many biological processes. Our previous studies on integrated succinylome profiles found that succinylation and acetylation lev-els of FEN1 would change under different conditions. Succinylation at FEN1 Lys200 site results in the accumulation of damaged DNA and in-creased susceptibility to fork-stalling agents. The interplay with other forms of modification could affects its protein interaction affinity and thus con-tribute to genome stability.
OBJECTIVE: This article studied the biological role of FEN1 by acyl modification in HeLa cells.
METHOD: In order to explore the function of FEN1 acylation in cells, we mimicked the presence or ab-sence of acetylation or succinylation by mutating key amino acids to glutam-ic acid and glutamine. We carried out a series of experiments including cell cycle, MTS, enzyme kinetics measurements, immunofluorescence and so on.
RESULTS: The absence of acylation of FEN1 leads to the blocked cell cycle process and the reduced efficiency of FEN1 on its DNA substrates, affect-ing the interaction of FEN1 with both repair and replication related proteins and thus its role in the repair of DNA damage.
CONCLUSION: We have veri-fied acyl groups could modify Lys125, Lys252 and Lys254 of FEN1. Acyl-ation level of these three is important for enzyme activity, cell proliferation and DNA damage response, thus contributing to genome stability.

Keywords:  DNA repair; FEN1; acetylation; genome stability; posttranslational modification; succinylation

DOI:  https://doi.org/10.2174/0929866530666230529152209