bims-toxgon Biomed News
on Toxoplasma gondii metabolism
Issue of 2022‒10‒30
sixteen papers selected by
Lakesh Kumar
BITS Pilani

  1. Trends Parasitol. 2022 Oct 25. pii: S1471-4922(22)00255-0. [Epub ahead of print]
      Using a CRISPR/Cas9-based method, Wincott et al. generated a stable, complex Toxoplasma gondii population composed of 96 barcoded clonal lineages. By tracking the population structure in vivo, they determine that - contrary to expectations - the pathway to infecting the brain is widely permissive for T. gondii.
    Keywords:  Toxoplasma gondii; blood–brain barrier; dissemination; genetic bottleneck; molecular barcode
  2. Trends Parasitol. 2022 Oct 24. pii: S1471-4922(22)00219-7. [Epub ahead of print]
      The mitochondrial respiratory chain is an essential pathway in most studied eukaryotes due to its roles in respiration and other pathways that depend on mitochondrial membrane potential. Apicomplexans are unicellular eukaryotes whose members have an impact on global health. The respiratory chain is a drug target for some members of this group, notably the malaria-causing Plasmodium spp. This has motivated studies of the respiratory chain in apicomplexan parasites, primarily Toxoplasma gondii and Plasmodium spp. for which experimental tools are most advanced. Studies of the respiratory complexes in these organisms revealed numerous novel features, including expansion of complex size. The divergence of apicomplexan mitochondria from commonly studied models highlights the diversity of mitochondrial form and function across eukaryotic life.
    Keywords:  Apicomplexa; F(o)-F(1) ATP synthase; Plasmodium; Toxoplasma; mitochondrial electron transport chain; mitochondrion
  3. Animals (Basel). 2022 Oct 20. pii: 2858. [Epub ahead of print]12(20):
      Toxoplasma gondii (T. gondii) is responsible for severe human and livestock diseases, huge economic losses, and adversely affects the health of the public and the development of animal husbandry. Glycosylation is a common posttranslational modification of proteins in eukaryotes, and N-glycosylation is closely related to the biological functions of proteins. However, glycosylation alterations in the feline small intestine following T. gondii infection have not been reported. In this study, the experimental group was intragastrically challenged with 600 brain cysts of the Prugniuad (Pru) strain that were collected from infected mice. The cats' intestinal epithelial tissues were harvested at 10 days post-infection and then sent for protein glycosylation analysis. High-performance liquid chromatography coupled to tandem mass spectrometry was used to analyze the glycosylation alterations in the small intestine of cats infected with T. gondii. The results of the present study showed that 56 glycosylated peptides were upregulated and 37 glycosylated peptides were downregulated in the feline small intestine infected by T. gondii. Additionally, we also identified eight N-glycosylated proteins of T. gondii including eight N-glycopeptides and eight N-glycosylation sites. The protein A0A086JND6_TOXGO (eEF2) and its corresponding peptide sequence were identified in T. gondii infection. Some special GO terms (i.e., cellular process and metabolic process, cell and cell part, and catalytic activity) were significantly enriched, and the Clusters of Orthologous Groups of proteins (COG) function prediction results showed that posttranslational modification, protein turnover, and chaperones (11%) had the highest enrichment for T. gondii. Interestingly, eEF2, a protein of T. gondii, is also involved in the significantly enriched T. gondii MAPK pathway. The host proteins ICAM-1 and PPT1 and the endoplasmic reticulum stress pathway may play an important role in the glycosylation of Toxoplasma-infected hosts. This is the first report showing that T. gondii oocysts can undergo N-glycosylation in the definitive host and that eEF2 is involved, which may provide a new target for T. gondii detection to prevent the spread of T. gondii oocysts in the future.
    Keywords:  N-glycosylation; Toxoplasma gondii; glycosylation; oocysts
  4. Pathogens. 2022 Oct 03. pii: 1142. [Epub ahead of print]11(10):
      Toxoplasma gondii is one of the most common protozoan parasites and is widely present in all warm-blooded animals. Although clinical disease is uncommon, some species, including ring-tailed lemurs (Lemur catta), have been found to develop acute and lethal toxoplasmosis. The aim of this study was to describe the pathologic, immunohistochemical, serological, and molecular findings of an outbreak of fatal toxoplasmosis in three captive ring-tailed lemurs in Central Italy in 2009. The animals died acutely within few days. The necropsy was immediately performed; necrotic lesions in the spleen, liver, and kidney, as well as interstitial pneumonia, were found histologically. All animals had high titers of anti-T. gondii-specific antibodies (1:1280 IgM and 1:640 IgG) according to a modified agglutination test (MAT) and immunohistochemistry showed scattered tachyzoites in the target organs. Diagnosis was confirmed by PCR and clonal type II was identified. In addition, the seven co-habiting lemurs were seronegative. This paper reports the first outbreak of acute disseminated toxoplasmosis in captive ring-tailed lemurs in Italy. These findings confirm the high susceptibility of this endangered species to toxoplasma infection, which may be considered a further threat to captive population viability.
    Keywords:  IHC; MAT; Toxoplasma gondii; fatal toxoplasmosis; genotyping; lemurs; zoo animals
  5. Parasit Vectors. 2022 Oct 27. 15(1): 393
      BACKGROUND: Women in early pregnancy infected by Toxoplasma gondii may have severe adverse pregnancy outcomes, such as spontaneous abortion and fetal malformation. The inhibitory molecule T cell immunoglobulin and mucin domain 3 (Tim-3) is highly expressed on decidual dendritic cells (dDCs) and plays an important role in maintaining immune tolerance. However, whether T. gondii infection can cause dDC dysfunction by influencing the expression of Tim-3 and further participate in adverse pregnancy outcomes is still unclear.METHODS: An abnormal pregnancy model in Tim-3-deficient mice and primary human dDCs treated with Tim-3 neutralizing antibodies were used to examine the effect of Tim-3 expression on dDC dysfunction after T. gondii infection.
    RESULTS: Following T. gondii infection, the expression of Tim-3 on dDCs was downregulated, those of the pro-inflammatory functional molecules CD80, CD86, MHC-II, tumor necrosis factor-α (TNF-α), and interleukin-12 (IL-12) were increased, while those of the tolerant molecules indoleamine 2,3-dioxygenase (IDO) and interleukin-10 (IL-10) were significantly reduced. Tim-3 downregulation by T. gondii infection was closely associated with an increase in proinflammatory molecules and a decrease in tolerant molecules, which further resulted in dDC dysfunction. Moreover, the changes in Tim-3 induced by T. gondii infection further reduced the secretion of the cytokine IL-10 via the SRC-signal transducer and activator of transcription 3 (STAT3) pathway, which ultimately contributed to abnormal pregnancy outcomes.
    CONCLUSIONS: Toxoplasma gondii infection can significantly downregulate the expression of Tim-3 and cause the aberrant expression of functional molecules in dDCs. This leads to dDC dysfunction, which can ultimately contribute to abnormal pregnancy outcomes. Further, the expression of the anti-inflammatory molecule IL-10 was significantly decreased by Tim-3 downregulation, which was mediated by the SRC-STAT3 signaling pathway in dDCs after T. gondii infection.
    Keywords:  Adverse pregnancy outcome; Decidual dendritic cells; Maternal–fetal tolerance; T cell immunoglobulin and mucin domain-containing protein 3; Toxoplasma gondii
  6. Int Immunopharmacol. 2022 Oct 25. pii: S1567-5769(22)00832-3. [Epub ahead of print]113(Pt A): 109348
      Roflumilast, a phosphodiesterase 4-inhibitor (PDE-4), shows immunomodulatory and anti-inflammatory properties. It modulates cAMP and TNF-α levels that play a role in the differentiation of Toxoplasma gondii (T. gondii) tachyzoite to bradyzoite stage. Thus, the potential effect of Roflumilast on the tachyzoite-bradyzoite transition in Me49 murine toxoplasmosis using 36 female Swiss Webster mice was studied. The mice were divided into six equal groups; normal control, infected control, two groups treated earlier with Roflumilast at 5 mg/kg and 10 mg/kg, and two groups treated later with Roflumilast at 5 mg/kg and 10 mg/kg. The T. gondii infection was assessed by cyst count and size, measuring serological levels of TNF-α and IL-12 using ELISA kits, brain immunohistochemistry examination of INF-γ and iNOS expression and histopathological examinations of the brain, liver, and spleen. The early-Roflumilast treated group at 10 mg/kg showed a statistically significant of reduction of T. gondii cyst count, size, and IL-12 level. In contrast, TNF-α levels were lower in both the early-Roflumilast treated groups. IFN-γ and iNOS expression showed non-significant changes in the different Roflumilast treated groups associated with mild inflammatory reactions in the brain, liver, and spleen tissues of the early-Roflumilast treated groups that were statistically significant (p < 0.05). This study showed that the earlier treated group at 10 mg/kg halted better tachyzoite-bradyzoite transition than the other groups. The results indicated Roflumilast to be promising for toxoplasmosis control.
    Keywords:  Bradyzoite; IL-12; INF-γ; Roflumilast; TNF-α; Toxoplasma gondii; iNOS
  7. Vaccines (Basel). 2022 Sep 22. pii: 1588. [Epub ahead of print]10(10):
      Heterologous immunization is garnering attention as a promising strategy to improve vaccine efficacy. Vaccines based on recombinant baculovirus (rBV) and virus-like particle (VLP) are safe for use, but heterologous immunization studies incorporating these two vaccine platforms remain unreported to date. Oral immunization is the simplest, most convenient, and safest means for mass immunization. In the present study, mice were immunized with the Toxoplasma gondii rhoptry protein 18 (ROP18)-expressing rBVs (rBVs-ROP18) and VLPs (VLPs-ROP18) via oral, intranasal, and intramuscular (IM) routes to evaluate the protection elicited against the intracellular parasite T. gondii ME49 strain. Overall, boost immunization with VLPs-ROP18 induced a significant increase in T. gondii-specific antibody response in all three immunization routes. Parasite-specific mucosal and cerebral antibody responses were observed from all immunization groups, but the highest mucosal IgA response was detected from the intestines of orally immunized mice. Antibody-secreting cell (ASC), CD8+ T cell, and germinal center B cell responses were strikingly similar across all three immunization groups. Oral immunization significantly reduced pro-inflammatory cytokine IL-6 in the brains as well as that by IN and IM. Importantly, all of the immunized mice survived against lethal challenge infections where body weight loss was negligible from all three immunizations. These results demonstrated that protection induced against T. gondii by oral rBV-VLP immunization regimen is just as effective as IN or IM immunizations.
    Keywords:  ROP18; Toxoplasma gondii; baculovirus; vaccine; virus-like particle
  8. Heliyon. 2022 Oct;8(10): e11091
      The AMP-activated protein kinase (AMPK) is a cellular sensor of energetics and when activated in skeletal muscle during contraction can impart changes in skeletal muscle metabolism. Therapeutics that selectively activate AMPK have been developed to lower glucose levels through increased glucose disposal rates as an approach to abrogate the hyperglycemic state of diabetes; however, the metabolic fate of glucose following AMPK activation remains unclear. We have used a combination of in vivo evaluation of glucose homeostasis and ex vivo skeletal muscle incubation to systematically evaluate metabolism following pharmacological activation of AMPK with PF-739, comparing this with AMPK activation through sustained intermittent electrical stimulation of contraction. These methods to activate AMPK result in increased glucose uptake but divergent metabolism of glucose: pharmacological activation results in increased glycogen accumulation while contraction-induced glucose uptake results in increased lactate formation and glucose oxidation. These results provide additional evidence to support a role for AMPK in control of skeletal muscle metabolism and additional insight into the potential for AMPK stimulation with small molecule direct activators.
    Keywords:  AMPK; Metabolism; Pharmacology; Skeletal muscle
  9. Mol Cell. 2022 Oct 16. pii: S1097-2765(22)00954-6. [Epub ahead of print]
      Pyruvate carboxylase (PC) catalyzes the two-step carboxylation of pyruvate to produce oxaloacetate, playing a key role in the maintenance of metabolic homeostasis in cells. Given its involvement in multiple diseases, PC has been regarded as a potential therapeutic target for obesity, diabetes, and cancer. Albeit acetyl-CoA has been recognized as the allosteric regulator of PC for over 60 years, the underlying mechanism of how acetyl-CoA induces PC activation remains enigmatic. Herein, by using time-resolved cryo-electron microscopy, we have captured the snapshots of PC transitional states during its catalytic cycle. These structures and the biochemical studies reveal that acetyl-CoA stabilizes PC in a catalytically competent conformation, which triggers a cascade of events, including ATP hydrolysis and the long-distance communication between the two reactive centers. These findings provide an integrated picture for PC catalysis and unveil the unique allosteric mechanism of acetyl-CoA in an essential biochemical reaction in all kingdoms of life.
    Keywords:  acetyl-CoA; allosteric regulation; biochemical reactions; cancer; diabetes; metabolism; pyruvate carboxylase
  10. Subcell Biochem. 2022 ;100 393-426
      Lysine acetylation is the second most well-studied post-translational modification after phosphorylation. While phosphorylation regulates signaling cascades, one of the most significant roles of acetylation is regulation of chromatin structure. Acetyl-coenzyme A (acetyl-CoA) serves as the acetyl group donor for acetylation reactions mediated by lysine acetyltransferases (KATs). On the other hand, NAD+ serves as the cofactor for lysine deacetylases (KDACs). Both acetyl-CoA and NAD+ are metabolites integral to energy metabolism, and therefore, their metabolic flux can regulate the activity of KATs and KDACs impacting the epigenome. In this chapter, we review our current understanding of how metabolic pathways regulate lysine acetylation in normal and cancer cells.
    Keywords:  Acetyl-CoA; Acetylation; Epigenome; Metabolic reprogramming; Metabolism; NAD+; Oral cancer
  11. Subcell Biochem. 2022 ;100 239-267
      The regulation of gene expression is a dynamic process that is influenced by both internal and external factors. Alteration in the epigenetic profile is a key mechanism in the regulation process. Epigenetic regulators, such as enzymes and proteins involved in posttranslational modification (PTM), use different cofactors and substrates derived from dietary sources. For example, glucose metabolism provides acetyl CoA, S-adenosylmethionine (SAM), α- ketoglutarate, uridine diphosphate (UDP)-glucose, adenosine triphosphate (ATP), nicotinamide adenine dinucleotide (NAD+), and fatty acid desaturase (FAD), which are utilized by chromatin-modifying enzymes in many intermediary metabolic pathways. Any alteration in the metabolic status of the cell results in the alteration of these metabolites, which causes dysregulation in the activity of chromatin regulators, resulting in the alteration of the epigenetic profile. Such long-term or repeated alteration of epigenetic profile can lead to several diseases, like cancer, insulin resistance and diabetes, cognitive impairment, neurodegenerative disease, and metabolic syndromes. Here we discuss the functions of key nutrients that contribute to epigenetic regulation and their role in pathophysiological conditions.
    Keywords:  Carbon metabolism; Dietary metabolites; Histone deacetylation; Metabolic disorders; Metabolic pathways; Vitamins
  12. Metabolism. 2022 Oct 21. pii: S0026-0495(22)00216-5. [Epub ahead of print] 155338
      BACKGROUND: Homeostasis of autophagy under normal conditions and nutrient stress is maintained by adaptive activation of regulatory proteins. However, the protein-lipid crosstalk that modulates the switch from suppression to activation of autophagy initiation is largely unknown.RESULTS: Here, we show that human diazepam-binding inhibitor (DBI), also known as acyl-CoA binding protein (ACBP), binds to phosphatidylethanolamine of the phagophore membrane under nutrient-rich growth conditions, leading to inhibition of LC3 lipidation and suppression of autophagy initiation. Specific residues, including the conserved tyrosine residues of DBI, interact with phosphatidylethanolamine to stabilize the later molecule in the acyl-CoA binding cavity of the protein. Under starvation, phosphorylation of serine-21 of DBI mediated by the AMP-activated protein kinase results in a drastic reduction in the affinity of the protein for phosphatidylethanolamine. The release of serine-21 phosphorylated DBI from the phagophore upon nutrient starvation restores the high LC3 lipidation flux and maturation of the phagophore to autophagosome.
    CONCLUSION: DBI acts as a strategic barrier against overactivation of phagophore maturation under nutrient-rich conditions, while triggering autophagy under nutrient-deficient conditions.
    Keywords:  Autophagy; Diazepam-binding inhibitor; Nutrient stress; Phosphorylation; Protein-lipid binding
  13. J Microbiol. 2022 Oct 24.
      Acetyl-CoA synthetase (ACS) is the enzyme that irreversibly catalyzes the synthesis of acetyl-CoA from acetate, CoA-SH, and ATP via acetyl-AMP as an intermediate. In this study, we demonstrated that AcsA1 (MSMEG_6179) is the predominantly expressed ACS among four ACSs (MSMEG_6179, MSMEG_0718, MSMEG_3986, and MSMEG_5650) found in Mycobacterium smegmatis and that a deletion mutation of acsA1 in M. smegmatis led to its compromised growth on acetate as the sole carbon source. Expression of acsA1 was demonstrated to be induced during growth on acetate as the sole carbon source. The acsA1 gene was shown to be negatively regulated by Crp1 (MSMEG_6189) that is the major cAMP receptor protein (CRP) in M. smegmatis. Using DNase I footprinting analysis and site-directed mutagenesis, a CRP-binding site (GGTGA-N6-TCACA) was identified in the upstream regulatory region of acsA1, which is important for repression of acsA1 expression. We also demonstrated that inhibition of the respiratory electron transport chain by inactivation of the major terminal oxidase, aa3 cytochrome c oxidase, led to a decrease in acsA1 expression probably through the activation of CRP. In conclusion, AcsA1 is the major ACS in M. smegmatis and its gene is under the negative regulation of Crp1, which contributes to some extent to the induction of acsA1 expression under acetate conditions. The growth of M. smegmatis is severely impaired on acetate as the sole carbon source under respiration-inhibitory conditions.
    Keywords:  Mycobacterium smegmatis; acetate; acetyl-CoA synthetase; cAMP-receptor protein; gene regulation
  14. Eur J Med Chem. 2022 Oct 04. pii: S0223-5234(22)00714-0. [Epub ahead of print]244 114812
      The increasing resistance of Toxoplasma gondii to drugs and side effects of therapy indicate that specific treatment for these parasites is still needed. The 4-arylthiosemicarbazide derivatives seem to be a solution to this challenge because they have low cytotoxicity against host cells and high anti-T. gondii activity. The molecular mechanism for these compounds is related to the inhibition of tyrosine amino acids involved in the proliferation and parasitophorous vacuole formation. The pharmacokinetic analysis shows that 1-(4-Methylimidazol-5-oyl)-4-(4-nitrophenyl)thiosemicarbazide and 4-(3-Iodophenyl)-1-(4-methylimidazol-5-oyl)thiosemicarbazide administered intragastrically pass into the bloodstream and cross the blood-brain barrier, and the absorption of both compounds is first-order absorption. Toxicity analysis shows that our derivatives possess lower toxicity than the routinely used drugs trimethoprim, sulfadiazine and pyrimethamine, as was observed in the level of liver enzymes and creatinine. Both derivatives are highly potent antiparasitic agents against T. gondii, prolonged survival and cure parasite-infected mice. Additionally, significant reductions in cyst formation in the brain and heart were observed, but the highest decreases were noted in muscle and the level of bradyzoites was similar to these observed in mice treated with commercially used drugs. Collectively, the obtained results support the conclusion that both compounds are highly efficacious in a mouse model of acute and chronic toxoplasmosis.
    Keywords:  Acute toxoplasmosis; Chronic toxoplasmosis; Creatinine; Interferon; Liver
  15. J Cell Biol. 2022 Dec 05. pii: e202208103. [Epub ahead of print]221(12):
      The mechanistic target of rapamycin complex 1 (mTORC1), a multi-subunit protein kinase complex, interrogates growth factor signaling with cellular nutrient and energy status to control metabolic homeostasis. Activation of mTORC1 promotes biosynthesis of macromolecules, including proteins, lipids, and nucleic acids, and simultaneously suppresses catabolic processes such as lysosomal degradation of self-constituents and extracellular components. Metabolic regulation has emerged as a critical determinant of various cellular death programs, including apoptosis, pyroptosis, and ferroptosis. In this article, we review the expanding knowledge on how mTORC1 coordinates metabolic pathways to impinge on cell death regulation. We focus on the current understanding on how nutrient status and cellular signaling pathways connect mTORC1 activity with ferroptosis, an iron-dependent cell death program that has been implicated in a plethora of human diseases. In-depth understanding of the principles governing the interaction between mTORC1 and cell death pathways can ultimately guide the development of novel therapies for the treatment of relevant pathological conditions.
  16. Subcell Biochem. 2022 ;100 337-360
      Sirtuin 6 (SIRT6) is a member of the mammalian sirtuin family with deacetylase, deacylase, and mono-ADP-ribosyl-transferase activities. It is a multitasking chromatin-associated protein regulating different cellular and physiological functions in cells. Specifically, SIRT6 dysfunction is implicated in several aging-related human diseases, including cancer. Studies indicate that SIRT6 has a tumor-specific role, and it is considered a tumor suppressor as well as a tumor growth inducer, depending on the type of cancer. In this chapter, we review the role of SIRT6 in metabolism, genomic stability, and cancer. Further, we provide an insight into the interplay of the tumor-suppressing and oncogenic roles of SIRT6 in cancer. Additionally, we discuss the use of small-molecule SIRT6 modulators as potential therapeutics.
    Keywords:  Aging; Cancer; Genome instability; Metabolism; SIRT6