bims-toxgon Biomed News
on Toxoplasma gondii metabolism
Issue of 2025–07–20
sixteen papers selected by
Lakesh Kumar, BITS Pilani
  1. Ferroptosis is important for Toxoplasma gondii replication and virulence in vitro and in vivo.
  2. The m6A demethylase FTO regulates TNF-α expression in human macrophages following Toxoplasma gondii infection.
  3. The early human interferon gamma response to Toxoplasma gondii is driven by Vγ9Vδ2 T-cell sensing of host phosphoantigens and subsequent NK-cell activation.
  4. Toxoplasma effector Tg WIP hijacks dendritic cell actin and motility via Nck1/Grb2 and the WAVE complex.
  5. Reinterpreting the effects of α-tubulin K40 acetylation on microtubule stability and cellular functions.
  6. Glycosylation of Plasmodium falciparum TRAP is essential for infection of mosquito salivary glands.
  7. The apicoplast transportome of the malaria parasite.
  8. Two novel phosphatidylinositol phosphate-binding proteins are identified in Neospora caninum with NcPX1 playing a crucial role.
  9. DHX9-mediated epigenetic silencing of BECN1 contributes to impaired autophagy and tumor progression in breast cancer via recruitment of HDAC5.
  10. Dynamic assembly of malate dehydrogenase-citrate synthase multienzyme complex in the mitochondria.
  11. Novel insights into the ULK2-FIP200-AMPK-mediated regulation of autophagy and BCR::ABL degradation in chronic myeloid leukemia.
  12. Toxoplasma gondii Coinfection in HIV-Positive Patients at a Tertiary Care Hospital.
  13. Untargeted Metabolomics of Epimastigote Forms of Trypanosoma cruzi.
  14. Sirtuins and their role in ovarian aging-related fibrosis predisposing to ovarian cancer.
  15. Lipids: Driving Forces in the Underlying Biology of Carcinogenesis.
  16. [Research progress on macrophage metabolic reprogramming in ocular diseases].
  1. Virulence. 2025 Dec;16(1): 2530164
    Ferroptosis is important for Toxoplasma gondii replication and virulence in vitro and in vivo.
    Ling-Yu Li, Chun-Xue Zhou, Bing Han, Hany M Elsheikha, Hui-Jie Qiu, Xu-Dian An, Ting Zeng, Dai-Ang Liu, Qing Yang, Xing-Quan Zhu, Huai-Yu Zhou.
      The protozoan parasite T. gondii employs intricate mechanisms to exploit host cells while sustaining their viability, yet its interaction with ferroptosis - an iron-dependent cell death driven by lipid peroxidation - remains poorly defined. Here, we show T. gondii infection induces ferroptotic hallmarks in RAW264.7 macrophages, including elevated lactate dehydrogenase release, labile Fe2 + accumulation, reactive oxygen species (ROS) generation, and lipid peroxidation. Molecular analyses revealed infection-induced downregulation of ferroptosis suppressor GPX4 and upregulation of pro-ferroptotic ACSL4 in macrophages and mice. Mechanistically, the SLC7A11/GPX4 axis governed parasite growth: knockdown of these genes promoted T. gondii replication, whereas overexpression restricted proliferation. Pharmacological studies showed ferroptosis inhibitor Fer-1 suppressed intracellular parasite proliferation. Notably, GPX4 inhibitor RSL3 exhibited context-dependent effects: pre-infection treatment enhanced replication, while post-infection administration inhibited growth. Direct RSL3 exposure induced time-dependent growth arrest in extracellular tachyzoites, associated with disrupted transcriptomes, increased lipid ROS, and downregulated parasite antioxidant genes (TgPRX2, TgTPX1/2, TgNXN), indicating redox homoeostasis impairment. In vivo murine studies corroborated this biphasic effect: therapeutic RSL3 administration post-infection significantly reduced parasite burdens across multiple organs (spleen, liver, kidney, brain) and improved survival rates, while prophylactic pretreatment exacerbated disease progression. We propose RSL3 exerts direct parasiticidal effects via oxidative damage but also enables early nutrient acquisition from ferroptosis-compromised host cells. These findings establish ferroptosis as a critical node in T. gondii pathogenesis, highlighting the parasite's hijacking of host iron-lipid metabolism. The dual role of ferroptosis regulators underscores the host-pathogen metabolic complexity and positions the SLC7A11/GPX4 axis as a promising therapeutic target.
    Keywords:  GPX4; RSL3; T. gondii; ferroptosis; mouse
    DOI:  https://doi.org/10.1080/21505594.2025.2530164
  2. PLoS Negl Trop Dis. 2025 Jul 15. 19(7): e0013289
    The m6A demethylase FTO regulates TNF-α expression in human macrophages following Toxoplasma gondii infection.
    Min Qin, Nan Gao, Jierui Sun, Shuqing Lin, Tingting Hu, Xinjian Liu, Rong Zhang, Yong Wang, Jingfan Qiu.
      Toxoplasma gondii (T. gondii) is an opportunistic parasite. After infection, macrophages finely regulate the immune response to restrict parasite proliferation. It is well-known that N6-methyladenosine (m6A) plays a critical role in fine-tuning gene expression. To investigate whether m6A modification is involved in regulating the anti-infection immune response in human macrophages against T. gondii, this study utilized T. gondii tachyzoites from the RH strain to infect human THP-1 macrophages. qPCR and ELISA results show that T. gondii infection mounted the expression of TNF-α, IL-1β, and IL-6. Transcriptomic data suggest that the infection of T. gondii induced differential gene expression in pathways associated with TNF signaling and cytokine-cytokine receptor interaction. Meanwhile, expression of m6A regulators were evaluated using qPCR and Western blotting. T. gondii infection increased the abundance of m6A demethylase FTO and methyltransferase WTAP. Joint analysis of RNA-seq and m6A-seq data was utilized for enriching differentially expressed genes (DEGs) with significantly altered m6A modifications. Intriguingly, following T. gondii infection, the m6A levels of DEGs associated with toxoplasmosis, TNF signaling pathway, and NF-κB signaling pathway were significantly different. The m6A-IP-qPCR assay further confirmed that T. gondii infection led to the decrease in the levels of m6A modification in the 3'UTR and 5'UTR regions of TNF-α mRNA. Knocking-down of FTO retarded the infection induced-decrease in the levels of m6A modification in TNF-α transcripts, accompanied by dampened immune response and uncontrolled T. gondii proliferation. Furthermore, the YTHDF2 RIP assay indicates that T. gondii infection remarkably weakened the binding of YTHDF2 to TNF-α mRNA, which could mount TNF-α expression by inhibiting the degradation of TNF-α mRNA. Overall, these findings suggest that m6A plays a role in the T. gondii infection-induced immune response in human macrophages, uncovering a new molecular mechanism for the regulation of TNF-α expression from an epitranscriptomic aspect.
    DOI:  https://doi.org/10.1371/journal.pntd.0013289
  3. bioRxiv. 2025 Jun 17. pii: 2025.06.12.659293. [Epub ahead of print]
    The early human interferon gamma response to Toxoplasma gondii is driven by Vγ9Vδ2 T-cell sensing of host phosphoantigens and subsequent NK-cell activation.
    Felipe Rodriguez, Jeroen P J Saeij.
      Toxoplasma gondii is a globally prevalent intracellular parasite that infects ∼40 million Americans. The murine immune response to Toxoplasma relies on both toll-like receptor (TLR) 11/12 and immunity related GTPase-mediated (IRGs) responses, which humans lack, making it unclear how the human immune response detects and responds to the parasite. We investigated whether human Vγ9Vδ2 T cells, which detect phosphoantigens through the BTN3A1 receptor, shape the early immune response to the parasite. Using primary human peripheral blood mononuclear cells (PBMCs), we show that Vγ9Vδ2 T cells are activated by Toxoplasma -infected cells in a BTN3A1- dependent manner leading to secretion of interferon gamma (IFNγ) and tumor necrosis factor-alpha (TNFα). Additionally, these T cells potentiate IFNγ production by natural killer (NK) cells, likely via TNFα and interleukin (IL)-12 produced during infection. Active parasite invasion is required to stimulate the IFNγ response, and inhibition of the host mevalonate pathway, which limits the synthesis of the phosphoantigen isopentenyl pyrophosphate (IPP), attenuates the cytokine response, indicating Toxoplasma infection increases host phosphoantigens leading to Vγ9Vδ2 T cell activation. Our findings identify Vγ9Vδ2 T cells as key effectors that potentiate NK cells in the early human immune response to Toxoplasma , bridging innate and adaptive immunity in the absence of TLR11/12 signaling.
    DOI:  https://doi.org/10.1101/2025.06.12.659293
  4. bioRxiv. 2025 Jun 23. pii: 2025.06.22.660983. [Epub ahead of print]
    Toxoplasma effector Tg WIP hijacks dendritic cell actin and motility via Nck1/Grb2 and the WAVE complex.
    Pavel Morales, Daniel A Kramer, Caroline de Moraes de Siqueira, Abbigale J Brown, Lamba Omar Sangaré, Baoyu Chen, Jeroen P J Saeij.
      The intracellular parasite Toxoplasma gondii enhances its dissemination to distant organs by hijacking infected leukocytes via a Trojan Horse mechanism. Upon infecting dendritic cells (DCs), Toxoplasma induces a hypermigratory phenotype characterized by podosome dissolution and formation of F-actin stress fibers. We previously showed that these cytoskeletal changes depend on the effector protein Toxoplasma WAVE complex-interacting protein ( Tg WIP) secreted from parasites to infected leukocytes. Here, we identify the host adaptor proteins Non-catalytic region of tyrosine kinase adaptor protein 1 and 2 (Nck1/2) and Growth factor receptor-bound protein 2 (Grb2) as direct Tg WIP interactors. Tg WIP mainly uses two distinct proline-rich regions (PRRs) to interact with Nck1 and Grb2. Mutating these PRRs abrogates Tg WIP binding to Nck1 and Grb2 and diminishes podosome dissolution and DC hypermotility. Furthermore, we show that Tg WIP directly interacts with the actin nucleation promoting factor WAVE Regulatory Complex (WRC) via a WRC-interacting receptor sequence (WIRS). Disrupting this interaction also influences actin cytoskeletal remodeling and DC hypermotility. Collectively, our data reveal that Tg WIP directly interacts with multiple actin regulators, including Nck1, Grb2, and the WRC, to remodel the actin cytoskeleton of the host cells, elucidating a key mechanism that Toxoplasma exploits to enhance host cell migration and dissemination.
    DOI:  https://doi.org/10.1101/2025.06.22.660983
  5. J Cell Sci. 2025 Jul 15. pii: jcs263431. [Epub ahead of print]138(14):
    Reinterpreting the effects of α-tubulin K40 acetylation on microtubule stability and cellular functions.
    Yu-Ming Lu.
      Acetylation of α-tubulin at lysine 40 (K40) has been studied in many model organisms for decades, mainly by manipulating levels of deacetylase and acetyltransferase enzymes, such as the α-tubulin acetyltransferase MEC-17 (also known as ATAT1). Observations that acetylation accumulates in some long-lived microtubules and that MEC-17 is important for maintaining microtubule organization and key cellular functions have led to the prevailing view that K40 acetylation stabilizes and protects microtubules, although many questions about its precise function remain. Recent gene editing of endogenous α-tubulin and in vitro microtubule polymerization assays have indicated that K40 acetylation itself does not maintain microtubule structure as MEC-17 does, but rather negatively regulates specific aspects of microtubule dynamics (i.e. nucleation and shrinkage but not elongation) and slightly impairs neuronal extension. This Opinion article discusses multiple important studies on α-tubulin K40 acetylation that have shaped our understanding of its function since its discovery in the 1980s, with the aim of clarifying the actual role of this major tubulin post-translational modification.
    Keywords:  ATAT1; K40 acetylation; MEC-17; Microtubules; Tubulin acetylation; Tubulin post-translational modifications
    DOI:  https://doi.org/10.1242/jcs.263431
  6. bioRxiv. 2025 Jun 26. pii: 2025.06.26.658380. [Epub ahead of print]
    Glycosylation of Plasmodium falciparum TRAP is essential for infection of mosquito salivary glands.
    Priya Gupta, Nastaran Rezakhani, Lucia Pazzagli, Hardik Patel, Gigliola Zanghi, Mohd Kamil, Alexander Watson, Vladimir Vigdorovich, Nelly Camargo, Erin Knutson, D Noah Sather, Ashley M Vaughan, Kristian E Swearingen.
      The human malaria parasite Plasmodium falciparum ( Pf ) expresses ten thrombospondin type 1 repeat (TSR) domain-bearing proteins at different stages throughout its life cycle. TSRs can be modified by two types of glycosylation: O-fucosylation at conserved serine (S) or threonine (T) residues and C-mannosylation at conserved tryptophan (W) residues. Pf TRAP, which is expressed in mosquito-stage sporozoites, has one TSR domain that is O-fucosylated at Thr 256 and C-mannosylated at Trp 250 . We employed site-directed mutagenesis by CRISPR/Cas9 gene editing to generate two Pf TRAP glyco-null mutant parasites, Pf TRAP_T256A and Pf TRAP_W250F. The fitness of these mutant parasites across the life cycle was compared to the parental NF54 wild type as well as a Pf TRAP knockout line. The glyco-null parasites exhibited major fitness defects comparable to knockout: sporozoites were unable to productively colonize the salivary glands and were highly impaired in gliding motility and the ability to invade cultured human hepatocytes. Protein analysis revealed significantly reduced Pf TRAP abundance in the glyco-null mutants despite normal transcript levels. These findings demonstrate that glycosylation of Pf TRAP's TSR is critical for its proper expression and function, and underscores the importance of TSR glycosylation in the mosquito stage of the life cycle.
    DOI:  https://doi.org/10.1101/2025.06.26.658380
  7. Trends Parasitol. 2025 Jul 15. pii: S1471-4922(25)00180-1. [Epub ahead of print]
    The apicoplast transportome of the malaria parasite.
    Laura J Akkerman, Taco W A Kooij, Laura E de Vries.
      The apicoplast, a peculiar organelle of red algal origin surrounded by four membranes, is important for several metabolic processes in the malaria parasite, including isopentenyl pyrophosphate (IPP) and coenzyme A (CoA) biosynthesis. Transporters are required to provide substrates and export products for these metabolic pathways and are therefore excellent novel drug targets. On the basis of known apicoplast pathways, we discuss which functions are expected to be fulfilled by the Plasmodium apicoplast transportome, which comprises 11 confirmed and 17 putative apicoplast transporters identified to date. Facilitated by the development of new tools, we anticipate the discovery of key players of the apicoplast transportome, such as the IPP and CoA exporters, and the exploitation of these proteins as drug targets.
    Keywords:  Plasmodium; apicoplast; transporters
    DOI:  https://doi.org/10.1016/j.pt.2025.06.011
  8. Acta Trop. 2025 Jul 09. pii: S0001-706X(25)00209-8. [Epub ahead of print]268 107737
    Two novel phosphatidylinositol phosphate-binding proteins are identified in Neospora caninum with NcPX1 playing a crucial role.
    Yilei Zhang, Zhouchun Li, Yangfan Li, Yuhan Zhou, Guodong Xiao, Zhenxiao Lu, Qianming Xu, Congshan Yang.
      Neospora caninum is a unicellular parasite that causes neosporosis, a major contributor to reproductive disorders in livestock. Whether phosphatidylinositol phosphate (PIP)-binding proteins play a key role in the life cycle of the parasite is unknown. To investigate the functional roles of PIP-binding proteins in the intracellular parasitism and pathogenicity of N. caninum. We identified two novel PIP-binding proteins, NcPX1 and NcPH1, and evaluated their biological functions using CRISPR/Cas9-generated knockout strains. The Δncpx1 strain displayed reduced replication and invasion, along with altered liposomes accumulation in extracellular parasites and impaired expression of dense granule proteins (GRAs) and microneme proteins (MICs). In vivo assays revealed significantly prolonged survival in mice infected with the Δncpx1 strain, underscoring the role of NcPX1 in pathogenicity. Conversely, the Δncph1 strain showed negligible phenotypic changes. These findings establish NcPX1 as a key determinant of N. caninum infection and highlight PIP-binding proteins as potential therapeutic targets for controlling neosporosis.
    Keywords:  NcPH1; NcPX1; Neospora caninum; Phenotype; Phosphatidylinositol phosphate
    DOI:  https://doi.org/10.1016/j.actatropica.2025.107737
  9. Cell Death Dis. 2025 Jul 14. 16(1): 524
    DHX9-mediated epigenetic silencing of BECN1 contributes to impaired autophagy and tumor progression in breast cancer via recruitment of HDAC5.
    Ziyang Li, Fang Liu, Fengbei Li, Guopeng Zeng, Xin Wen, Jianan Ding, Jueyu Zhou.
      Autophagy is closely linked to tumorigenesis, progression and metastasis. DHX9 is a member of the DExD/H-box helicase family and plays important roles in transcription, translation, RNA editing and non-coding RNA synthesis. Mounting evidence demonstrates that aberrant expression of DHX9 is associated with the development and progression of several tumors. However, whether DHX9 regulates autophagy deficiency in breast cancer (BC) remains unknown. Herein, we found that DHX9 expression was frequently elevated in BC cells and tissues, which suggested poor survival. The viability and motility of BC cells were irritated by enhanced DHX9 expression. Meanwhile, reduced DHX9 expression postponed tumor development both in vitro and in vivo. Subsequent research revealed that DHX9 knockdown suppressed the activation of the mTOR signaling pathway and accelerated autophagic flux by promoting the formation of autophagosomes in BC cells. Mechanistically, DHX9 occupied the proximal promoter of BECN1 and repressed its transcription. DHX9-mediated BECN1 inhibition required histone deacetylase (HDAC) activity. HDAC5 was recruited to the nucleus and co-localized with DHX9 at the BECN1 promoter, mediating the deacetylation of histone H3 and ultimately inhibited BECN1 transcription. Importantly, the tumor-suppressive effect of DHX9 knockdown was reversed by BECN1 downregulation. In conclusion, the previously unrecognized significance of DHX9 in mediating the epigenetic silencing of BECN1, which is essential for autophagy and tumorigenesis, highlights its potential as an effective biomarker as well as a prospective therapeutic candidate for BC.
    DOI:  https://doi.org/10.1038/s41419-025-07847-y
  10. bioRxiv. 2025 Jun 20. pii: 2025.06.16.659985. [Epub ahead of print]
    Dynamic assembly of malate dehydrogenase-citrate synthase multienzyme complex in the mitochondria.
    Joy Omini, Inga Krassovskaya, Taiwo Dele-Osibanjo, Connor Pedersen, Toshihiro Obata.
      The tricarboxylic acid (TCA) cycle enzymes, malate dehydrogenase (MDH1) and citrate synthase (CIT1), form a multienzyme complex called 'metabolon' that channels intermediate, oxaloacetate, between the reaction centers of the enzymes. Since the MDH1-CIT1 metabolon enhances the pathway reactions in vitro, it is postulated to regulate the TCA cycle flux through dynamic assembly in response to cellular metabolic demands. Here, we demonstrated that yeast mitochondrial MDH1 and CIT1 dissociated when aerobic respiration was suppressed by the Crabtree effect and associated when the pathway flux was enhanced by acetate. Pharmacological TCA cycle inhibitions dissociated the complex, while electron transport chain inhibition enhanced the interaction. The multienzyme complex assembly was related to the mitochondrial matrix acidification and oxidation, as well as cellular levels of malate, fumarate, and citrate. These factors significantly affected the MDH1-CIT1 complex affinity in vitro. Especially the buffer pH significantly changed the MDH1-CIT1 affinity within the pH range between 6.0 and 7.0, which is observed in the mitochondrial matrix under physiological conditions. These results show a dynamic association and dissociation of a metabolon in the mitochondria and its relationship with pathway flux, supporting the metabolon's role in metabolic regulation. Multiple factors, including pH and metabolite availabilities, possibly regulate MDH1-CIT1 interaction.
    DOI:  https://doi.org/10.1101/2025.06.16.659985
  11. Biochem Biophys Res Commun. 2025 Jul 13. pii: S0006-291X(25)01058-7. [Epub ahead of print]778 152343
    Novel insights into the ULK2-FIP200-AMPK-mediated regulation of autophagy and BCR::ABL degradation in chronic myeloid leukemia.
    Daisuke Koyama, Naoki Yamamoto, Takaaki Abe, Kengo Suzuki, Yuki Sato, Manabu Suzuki, Naoya Shibayama, Takayuki Ikezoe.
      Chronic myeloid leukemia (CML) is a myeloproliferative neoplasm driven by the BCR::ABL fusion tyrosine kinase. AMP-activated protein kinase (AMPK) plays a pivotal role in regulating cellular energy homeostasis, ensuring an adequate ATP supply for CML cell proliferation. ULK1, a well-known AMPK substrate, is a critical serine/threonine kinase in the autophagy initiation complex. ULK2, a paralog of ULK1, shares approximately 50 % amino acid sequence homology and has been reported to function complementarily with ULK1. However, emerging evidence suggests that ULK2 also has unique functions distinct from those of ULK1. Public RNA sequencing data revealed that ULK2 expression is significantly lower in hematopoietic cells compared to other tissues. To explore the function of ULK2, we performed in vitro assays using 293FT cells, which endogenously express high levels of ULK2. Mass spectrometry analysis demonstrated that ULK2 forms a stable complex with FIP200, which in turn interacts specifically with the AMPK α1 and γ1 subunits. Furthermore, shRNA-mediated knockdown of ULK2 induced AMPK activation and promoted the cytoplasmic accumulation of ULK1 and FIP200, thereby inducing autophagy in CML cells. Although autophagy typically acts as a cytoprotective mechanism, in this context, the autophagy-dependent degradation of BCR::ABL induced cell death. These findings reveal a novel regulatory axis involving ULK2, FIP200, AMPK, and autophagy, suggesting a unique role for ULK2 in CML pathophysiology and offering potential therapeutic insights.
    Keywords:  AMPK; Autophagy; Chronic myeloid leukemia; FIP200; ULK2
    DOI:  https://doi.org/10.1016/j.bbrc.2025.152343
  12. Curr HIV Res. 2025 Jul 14.
    Toxoplasma gondii Coinfection in HIV-Positive Patients at a Tertiary Care Hospital.
    Muhammed Furkan Kürkçü, Ayfer Bakır, Semiha Berra Topsakal Kaba, İlknur Alkan Kuşabbi, Selma Usluca.
       INTRODUCTION: Toxoplasma gondii (T. gondii) can cause serious complications in both immunocompetent and immunosuppressed individuals. This study aims to assess the seroprevalence of T. gondii among HIV-positive individuals and to investigate its association with age, sex, CD4+ T cell count, HIV RNA levels, and hematological parameters.
    METHODS: This study included 247 HIV-positive individuals followed up at a tertiary care hospital between November 1, 2022, and November 30, 2024. We analyzed serum samples for T. gondii IgG antibodies using electrochemiluminescent microparticle immunoassay.
    RESULTS: The prevalence of T. gondii IgG seropositivity was found to be 32.8% (n=81; 95% CI: 26.9-39). The median age of seropositive individuals was 52 years (IQR: 42-61), which was significantly higher compared to seronegative individuals (p<0.001). The highest IgG seropositivity rate (66.7%) was observed in the 61-80 age group. Hemoglobin levels were significantly lower in IgG seropositive individuals (p=0.040). Logistic regression analysis indicated an increased risk of T. gondii infection with advancing age. The odds ratio for the 41-60 age group was 13.3 (95% CI: 1.6-106, p=0.02), while for the 61-80 age group, it was 28 (95% CI: 3.3-240, p=0.002).
    DISCUSSION: The seroprevalence of T. gondii in HIV-positive individuals was lower than both global and regional averages. Age was identified as an independent risk factor for T. gondii seropositivity. Additionally, hematological alterations associated with anemia were observed in seropositive individuals. Further large-scale, multi-center, and regionally representative studies are required to optimize T. gondii infection management and screening strategies in people living with HIV.
    CONCLUSION: These findings suggest that T. gondii infection in HIV-positive individuals increases with age and may be associated with anemia, highlighting the need for age-focused screening and management strategies.
    Keywords:  CD4+ T cell count; HIV; IgG; Toxoplasma gondii; risk factors.; seroprevalence
    DOI:  https://doi.org/10.2174/011570162X392027250704053346
  13. Bio Protoc. 2025 Jul 05. 15(13): e5368
    Untargeted Metabolomics of Epimastigote Forms of Trypanosoma cruzi.
    Michel Augusto Silva, Mario Izidoro, Bruno Souza Bonifácio, Sergio Schenkman.
      Trypanosoma cruzi, the causative agent of Chagas disease, faces significant metabolic challenges due to fluctuating nutrient availability and oxidative stress within its insect vector. Metabolomic techniques, such as gas chromatography-mass spectrometry (GC-MS), have been widely used to study the adaptive mechanisms of the parasite. This article describes a standardized method for the untargeted metabolomics analysis of T. cruzi epimastigote, covering parasite cultivation, sample deproteinization with methanol, metabolite extraction, derivatization with BSTFA, and GC-MS analysis. To ensure robustness and reproducibility, statistical analysis uses univariate tests, as well as multivariate approaches such as principal component analysis (PCA) and partial least squares (PLS) regression. The protocol offers a reliable and sensitive method to study metabolic responses in T. cruzi under environmental stress, with low biological variability and high reproducibility. Key features • GC-MS was used to conduct a standardized metabolomics investigation of Trypanosoma cruzi epimastigote, assuring reproducibility and minimum biological variability. • Includes sample deproteinization, metabolite extraction, and derivatization with BSTFA for accurate metabolite profiling under different biological conditions. • Employs robust statistical approaches (PCA, PLS) to investigate differences among experimental groups and detect significant alterations in metabolism. • Internal standards and multiple replicates ensure high sensitivity and repeatability, which is excellent for investigating metabolic processes in protozoan parasites.
    Keywords:  Bioinformatics; GC-MS; Metabolism; Metabolite; Metabolomics; Protozoan; T. cruzi
    DOI:  https://doi.org/10.21769/BioProtoc.5368
  14. NPJ Aging. 2025 Jul 15. 11(1): 65
    Sirtuins and their role in ovarian aging-related fibrosis predisposing to ovarian cancer.
    Arkadiusz Grzeczka, Agnieszka Skowronska, Sara Sepe, Mariusz T Skowronski, Paweł Kordowitzki.
      The pursuit of understanding early genetic or protein markers for ovarian aging has garnered considerable attention in the realm of reproductive medicine. Sirtuins (SIRTs) are a group of proteins that are NAD+-dependent, and thanks to their properties, they are able to change the acetylation profile of proteins and post-translationally modify their functions, too. Previous research provided evidence that SIRTs influence fibrosis levels in several organs. With regard to ovaries, fibrosis is one of the features of aged ovaries and also creates a metastasis-friendly environment, thus can also be a seedbed for the development of primary cancerous lesions. Ovarian cancer remains a formidable challenge in oncology due to its high prevalence, insidious onset, and frequent recurrence. Noteworthy, ovarian cancer is the seventh most common cancer among women and the eighth leading cause of cancer death worldwide. Ovarian fibrosis runs concurrently with the activation of TGF-β/Smads signaling, as well as inflammasome (NLRP3), nuclear factor kB (NFkB) and forkhead box O (FOXO) attenuation. Reduced levels of certain sirtuins resulting from decreased nicotinamide adenine dinucleotide (NAD + ) may underlie the dysregulation of the aforementioned signaling pathways and therefore represent a potential therapeutic target. This review elucidates the role of SIRTs in ovarian aging-related fibrosis as a process that predisposes to tumorigenesis.
    DOI:  https://doi.org/10.1038/s41514-025-00256-7
  15. ACS Pharmacol Transl Sci. 2025 Jul 11. 8(7): 1891-1918
    Lipids: Driving Forces in the Underlying Biology of Carcinogenesis.
    Sohini Chakraborty, Anudarshini Sathish Kumar, Satarupa Banerjee.
      Lipids, mainly composed of cholesterol, phospholipids, sphingolipids, triacylglycerides, and fatty acids, have vital functions within cells. Some lipids function as signaling molecules or secondary messengers and are cellular membranes' energy sources and structural elements. More research is being conducted on metabolic reprogramming as a hallmark of cancer. However, compared with the metabolism of glucose or glutamine, lipid metabolism in cancer has received less attention. There is increasing evidence that certain parts of the lipid metabolism are altered in cancer cells. The alterations could influence the quantity of lipids involved in signaling functions, affect the synthesis and breakdown of lipids necessary for maintaining energy homeostasis, and modify the availability of structural lipids critical for membrane formation. The term "lipid metabolic reprogramming" refers to modifications in the lipid metabolism that can impact cellular processes such as cell division, growth, proliferation, and the cell cycle, ultimately resulting in cancer. Furthermore, interactions between cancer cells and nearby immune cells via an altered lipid metabolism promote the development and spread of tumors. The most recent studies on the involvement of lipid metabolism in different cancers and associated hallmarks and lipids in various aspects of cancer therapeutics, which affect multiple facets of tumorigenesis, are described in this review.
    Keywords:  and; cancer hallmarks; lipid droplets and rafts; lipid reprogramming; lipogenic factors; therapeutics; treatment resistance
    DOI:  https://doi.org/10.1021/acsptsci.5c00170
  16. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2025 Aug;41(8): 744-749
    [Research progress on macrophage metabolic reprogramming in ocular diseases].
    Yun He, Junwen Ouyang, Qian Tan, Kai Hu.
      Macrophages are the crucial immune cells integral to host defense and the regulation of homeostasis, exhibiting remarkable plasticity across various tissues. Upon exposure to different stimuli, they can polarize into functional subsets. The reorganization process of cellular metabolism, known as metabolic reprogramming, involves the comprehensive adjustment of intracellular metabolites, enzymes, and metabolic pathways. Recent studies have revealed the critical role of metabolic reprogramming in shaping the phenotypes and functions of macrophages. Metabolism drives and regulates macrophages by generating bioenergy and biosynthetic precursors and by altering metabolites that affect gene expression and signal transduction. This review focuses on the immunomodulatory roles of key enzymes and specific products in major metabolic pathways, such as glucose metabolism, lipid metabolism and amino acid metabolism, in macrophages. Additionally, it will highlight recent advancements in targeting metabolic regulation of macrophages in the context of ocular diseases.
This page is being maintained by Thomas Krichel. It was last updated on 2025‒07‒20 at 10:10.