bims-toxgon 2026-08-02 papers

bims-toxgon

Biomed News

on Toxoplasma gondii metabolism

Issue of 2026–08–02
fifteen papers selected by
Lakesh Kumar, BITS Pilani

Structural basis for emetine inhibition of ribosome translocation in Toxoplasma gondii.
Toxoplasma TgATG2 works with TgATG9 and TgProp1 to drive autophagic flux for parasite extracellular survival and persistence.
Toxoplasma GRA8 engages the host ESCRT accessory protein ALG-2 and is necessary for parasite metabolic integrity.
AP2XI-3 is Critical for the Cell Cycle of Toxoplasma to Progress Through the G1 Phase.
From Insight to Impact: Spotlight on the Potential Implications of the Neuronal Nitric Oxide Synthase Isoenzyme (nNOS) in Experimental Chronic Toxoplasmosis.
Cryptosporidium sequentially remodels its single rhoptry into the host interface.
Competitive resource allocation drives asynchronous and rapid nuclear multiplication in the malaria parasite.
Cryptosporidium parvum CpARO marks rhoptry envelope remnants associated with feeder organelle formation.
Design, Synthesis, and Antitumor Activities of Novel Coumarin-Based Histone Deacetylase Inhibitors.
The role of autophagy in protozoan infections.
Acetylome Analysis in Vibrio vulnificus MO6-24/O Reveals Extensive Lysine Acetylation in Carbon Metabolism and Protein Synthesis Pathways: A Pilot Study.
Low-Glucose Metabolic Remodeling Drives Reversible Sequestration of Yeast S-Adenosylmethionine (AdoMet) Synthase.
Deciphering chromatin architecture and dynamics in Plasmodium falciparum using the nucDetective pipeline.
Temporal and spatial molecular heterogeneity of gastric cancer: Roles of post-translational modifications.
Label-Free Monitoring of Cancer-Associated Fibroblast Activation Using NAD(P)H Fluorescence Lifetime Imaging.

bioRxiv. 2026 Jul 24. pii: 2026.07.23.740346. [Epub ahead of print]

Structural basis for emetine inhibition of ribosome translocation in Toxoplasma gondii.

Wenzhao Dong, Fengrong Wang, Brenna A Saladin, Suada Leskaj, Ritam Neupane, Vern B Carruthers, Jailson Brito Querido.

Apicomplexan parasites, including Toxoplasma gondii and Plasmodium falciparum, are major human pathogens that cause toxoplasmosis and malaria, respectively. The existing structures of T. gondii translational machinery are from empty ribosomes that lack several key components, including ribosomal protein RACK1 (Receptor for Activated C Kinase 1). Here, we used cryo-electron microscopy (cryoEM) to determine high-resolution structures of T. gondii ribosomal complexes, including a translating 80S ribosome bound to mRNA and tRNA. These structures reveal that RACK1 occupies the conserved binding site on the 40S subunit observed in other eukaryotic ribosomes. We also determined the architecture of the ribosomal P-stalk and identified the ribosomal proteins uL10 and uL11, which were not observed in previous T. gondii ribosome structures. In addition, we determined structures of the 80S ribosome bound to mRNA, tRNA, and the translation inhibitor emetine in two distinct conformational states. These snapshots reveal two mechanisms by which emetine inhibits the translocation step of mRNA translation: either by dislodging the mRNA from the E-site of the ribosome or by acting as a molecular glue within the E-site, thereby stalling translocation. Together, these findings provide new insights into the molecular basis of protein synthesis in apicomplexan parasites and establish a structural framework for the development of future antiparasitic therapeutics.

DOI: https://doi.org/10.64898/2026.07.23.740346
bioRxiv. 2026 Jul 19. pii: 2026.07.16.739079. [Epub ahead of print]

Toxoplasma TgATG2 works with TgATG9 and TgProp1 to drive autophagic flux for parasite extracellular survival and persistence.

Silvia Masci, Pariyamon Thaprawat, Federica Piro, Tracey L Schultz, Vern B Carruthers, Manlio Di Cristina.

Chronic infection by Toxoplasma gondii depends on long-term survival of bradyzoites within tissue cysts, a parasite stage highly resistant to current therapies and a major barrier to eradication. Autophagy has emerged as critical pathway for bradyzoite persistence, yet the core machinery driving autophagosome formation in T. gondii remains poorly defined. Here, we identify TGME49_304630 as TgATG2, a previously uncharacterized, unusually large ATG2-like protein with conserved structural features of lipid-transfer factors. TgATG2 associates with TgATG9 and TgPROP1, key components of the parasite autophagy pathway, supporting its role in a membrane expansion complex required for autophagosome biogenesis. Using independent genetic knockouts, we show that TgATG2 is dispensable for intracellular tachyzoite replication but required for parasite fitness during extracellular stress and, most critically, for bradyzoite autophagy and viability. TgATG2 ablation disrupts autophagic activity in bradyzoites, causing progressive loss of viability and compromised cyst integrity. To overcome limitations of previous indirect assays, we developed a bradyzoite-specific dual-fluorescence TgATG8 reporter that quantitatively measures autophagic flux in T. gondii and confirmed TgATG2 as a major contributor. Importantly, TgATG2-deficient parasites are severely impaired during chronic infection in mice, with reduced brain cyst burdens, abnormal cyst morphology, and markedly diminished ex vivo bradyzoite viability. Together, these findings establish TgATG2 as a central component of the T. gondii autophagy machinery, demonstrate that autophagosome biogenesis is critical for parasite persistence in vivo, and define a molecular vulnerability and quantitative platform for targeting autophagy-dependent parasite survival.

DOI: https://doi.org/10.64898/2026.07.16.739079
bioRxiv. 2026 Jul 20. pii: 2026.07.20.739547. [Epub ahead of print]

Toxoplasma GRA8 engages the host ESCRT accessory protein ALG-2 and is necessary for parasite metabolic integrity.

Hargobinder Kaur, Rebekah B Guevara, Yolanda Rivera-Cuevas, Einar B Olafsson, Joshua Mayoral, Leonardo Augusto, Alfredo J Guerra, Romir Patel, Kevin P Bohannon, Jonathan Z Sexton, Phyllis I Hanson, Louis M Weiss, Vern B Carruthers.

The endosomal sorting complex required for transport (ESCRT) is a hetero-multimeric membrane-remodeling machinery essential for endosomal sorting, intraluminal vesicle formation, cytokinetic abscission, and membrane repair. ESCRT is also hijacked by some pathogens including the protozoan Toxoplasma gondii , which subverts it at the parasitophorous vacuole membrane to support parasite ingestion of host cytosolic proteins. Although the ESCRT accessory protein, ALG-2 is recruited to the parasitophorous vacuole, nothing was known about how this happens. Herein we identify the dense granule protein TgGRA8 as a key effector that recruits ALG-2 and ALIX to the parasitophorous vacuole. We show that TgGRA8 directly binds ALG-2 via conserved ALG-2-binding elements like those found in other ALG-2 interacting proteins including ALIX and SEC31A. Biochemical assays show high-affinity, Ca²⁺-dependent TgGRA8-ALG-2 binding, and structural modeling suggests TgGRA8 may assemble multivalently to coordinate multiple ALG-2 dimers, stabilizing ALG-2/ALIX recruitment through a non-canonical bridging mechanism. Conservation of these motifs among tissue cyst-forming coccidians implies a lineage-linked adaptation that supports infection by these parasites. Metabolomics further indicates that TgGRA8 loss disrupts amino acid, purine, and central carbon metabolism, like those seen in other ingestion deficient mutants. Together, these findings uncover a conserved, multivalent strategy by which Toxoplasma engages host ALG-2 to organize ESCRT at the parasitophorous vacuole, thereby coupling nutrient acquisition to metabolic fitness and exposing a novel agent for probing ESCRT biology.

DOI: https://doi.org/10.64898/2026.07.20.739547
Mol Microbiol. 2026 Jul 25.

AP2XI-3 is Critical for the Cell Cycle of Toxoplasma to Progress Through the G1 Phase.

Fuqiang Fan, Xiaoyan Yin, Anqi Liu, Lilan Xue, Jinling Chen, Yangfan Xu, Bang Shen.

  Toxoplasmosis caused by the Apicomplexan parasite Toxoplasma gondii is a significant health threat to immunocompromised individuals and newborns. This parasite has a complex life cycle that is well controlled to achieve optimal transmission and pathogenesis. Acute phase of the disease is caused by rapid proliferation of tachyzoites, which has a highly coordinated and tightly regulated cell cycle to allow parasite propagation. Tachyzoite cell cycle has five partially overlapping phases (G1, S, G2, M and C) with distinct gene expression patterns and cellular activities, yet the underlying regulatory mechanisms are not well understood. In this study, we show that the AP2 family transcription factor AP2XI-3 has important roles in regulating the cell cycle to progress through the G1 phase. Depletion of AP2XI-3 resulted in cell cycle retention at G1, leading to growth inhibition of tachyzoites. RNA-Seq and CUT&Tag analyses revealed that AP2XI-3 regulates the transcription of genes involved in nucleic acid metabolism, RNA biogenesis and processing, which are consistent with the cellular activities of G1 phase in preparing biomass for cell cycle progression. Moreover, AP2XI-3 binds to the conserved DNA motif, TRP-2, a sequence widely distributed in the promoters of genes that exhibit peak expression during the G1 phase. Together, these findings underscore the essential role of AP2XI-3 in the tight regulation of the G1 phase, highlighting its potential as a therapeutic target for drug development.

Keywords:  AP2; RNA biogenesis; cell cycle regulation; parasite division; transcriptional factors

DOI:  https://doi.org/10.1111/mmi.70099
Diseases. 2026 Jul 03. pii: 240. [Epub ahead of print]14(7):

From Insight to Impact: Spotlight on the Potential Implications of the Neuronal Nitric Oxide Synthase Isoenzyme (nNOS) in Experimental Chronic Toxoplasmosis.

Marwa Omar.

   BACKGROUND/OBJECTIVES: The coccidian protozoan Toxoplasma gondii (T. gondii) is among the most prevalent zoonotic parasites worldwide. Nitric oxide (NO) production by macrophages is considered a critical microbicidal mechanism against various intracellular pathogens, including T. gondii. While the role of the inducible nitric oxide synthase isoenzyme (iNOS) has been widely investigated in both acute and chronic T. gondii infections, the specific functions of the neuronal (nNOS) isotype in the antiparasitic immune response, particularly during chronic toxoplasmosis, remain largely uncovered. Hence, this report seeks to bridge the gap regarding the potential participation of nNOS in experimental chronic T. gondii infection.
METHODS: The study included 56 Swiss albino mice, equally allocated into four experimental groups: (G1) negative control, (G2) infected control, (G3) infected-L-arginine-treated, and (G4) infected-7-Nitroindazole-treated. All groups except (G1) were orally infected with the avirulent (ME49) T. gondii strain. Nine weeks post-infection, all mice were euthanized for parasitological, histopathological, immunohistochemical, and biochemical analyses.
RESULTS: The NO donor, L-arginine, induced a significant reduction in the number of T. gondii cysts, together with strong nNOS immunoreactivity in the brain sections of the treated mice. Conversely, the highest parasitic burden was observed following selective nNOS inhibition with 7-Nitroindazole, exacerbating parasite-induced pathology.
CONCLUSIONS: The neuronal isotype serves as a critical source of NO production during the chronic stage of T. gondii infection, thereby enhancing parasite elimination and contributing to host tissue protection.

Keywords:  7-Nitroindazole; L-arginine; Toxoplasma gondii; brain tissue; chronic toxoplasmosis; nNOS; nitric oxide

DOI:  https://doi.org/10.3390/diseases14070240
bioRxiv. 2026 Jul 21. pii: 2026.07.21.739819. [Epub ahead of print]

Cryptosporidium sequentially remodels its single rhoptry into the host interface.

Allison Cohen, Tim Bergner, Amandine Guérin, Maryse Lebrun, Michael Laue, Christian Klotz, Boris Striepen.

The apicomplexan parasite Cryptosporidium is a leading cause of diarrheal disease in young children. Within the intestinal epithelium, Cryptosporidium establishes a unique intracellular niche in the apical brush border of enterocytes. A structurally complex interface between host and parasite acts as a holdfast and enables protein and metabolite transport. How the parasite invades its host cell and builds the interface is poorly understood. Here, we reveal parasite invasion with high temporal and spatial resolution using rigorous molecular markers. We find that sequential discharge of specialized secretory organelles anchors the parasite within the host prior to internalization. The single Cryptosporidium rhoptry persists beyond its initial pre-invasion discharge, acting as a conduit for the secretion of multiple waves of effector proteins. Ultimately, the rhoptry membrane gives rise to the feeder organelle that separates host from parasite cytoplasm. These findings lead us to propose a mechanistic model of Cryptosporidium invasion and intracellular parasitism.

DOI: https://doi.org/10.64898/2026.07.21.739819
Nat Commun. 2026 07 27. pii: 7413. [Epub ahead of print]17(1):

Competitive resource allocation drives asynchronous and rapid nuclear multiplication in the malaria parasite.

Patrick Binder, Aistė Kudulytė, Severina Klaus, Thomas Höfer, Ulrich S Schwarz, Markus Ganter, Nils B Becker.

The unicellular malaria parasite Plasmodium falciparum proliferates within red blood cells of its human host, where it generates approximately 20 new parasites within a two-day developmental cycle. Before cellularization and release of the daughter cells, the nuclei multiply in a shared cytoplasm. In stark contrast to highly synchronized nuclear division cycles seen in other developing eukaryotes, Plasmodium nuclear cycles desynchronize rapidly. Combining live-cell imaging with biophysical modeling, we elucidate the mechanism of desynchronization and study its impact on parasite proliferation. We find that standard models of autonomous nuclear cycles cannot account for the experimental data, and therefore desynchronization requires nuclear coupling. Competition for a limiting pool of proteins needed for DNA replication explains the data, provided that they are allocated sequentially to individual nuclei. Sequential allocation can be achieved by reversible but stable association of the resources with DNA. Remarkably, the resultant asynchronous nuclear cycles accelerate parasite proliferation by minimizing idling times of the resource. This mechanism may be a general strategy to maximize proliferation in suboptimal growth conditions. Together, our findings identify nuclear cycle asynchrony as a resource-efficient means to achieve rapid proliferation.

DOI: https://doi.org/10.1038/s41467-026-75378-x
Virulence. 2026 Dec;17(1): 2707806

Cryptosporidium parvum CpARO marks rhoptry envelope remnants associated with feeder organelle formation.

Ying Zhang, Chenchen Wang, Xin Zhao, Xiaodong Wu, Jigang Yin, Guan Zhu.

  Apicomplexans encode a single armadillo repeat-only (ARO) protein, exemplified by TgARO and PfARO, that anchors to the rhoptry envelope through N-terminal acylation and supports rhoptry positioning through interaction with an ARO-interacting protein (AIP). These AROs organize rhoptries but are not known to be secreted during invasion. Here, we show that Cryptosporidium parvum ARO (CpARO) localizes to the rhoptry envelope by immunofluorescence assay, ultrastructural expansion microscopy, and structured illumination microscopy. During sporozoite invasion, CpARO-positive rhoptry envelope structures shorten and condense into a discrete punctum after content discharge. Residual rhoptry membrane structures are subsequently detected between the host cell F-actin pad and parasite nucleus in developing trophozoites, consistent with a contribution to nascent feeder organelle formation. In contrast to TgARO and PfARO, CpARO is also detected in secreted fractions during excystation, gliding, invasion, and intracellular development, with no evidence of nuclear localization. Recombinant CpARO binds host cells with high affinity (Kd = 0.189 μM). Although C. parvum encodes an AIP homolog, this protein localizes to the sporozoite cytoplasm rather than to rhoptries; we therefore designate it AIP-like protein (CpAIP-L). Antibodies against both CpARO and CpAIP-L were detected in sera from C. parvum-infected mice. These findings reveal functional divergence of Cryptosporidium ARO-AIP-related proteins and identify CpARO as both a rhoptry envelope marker and a secreted host-interacting factor with potential roles in host interaction and virulence.

Keywords:  Cryptosporidium parvum; armadillo repeat–only protein; feeder organelle formation; host–parasite interaction; rhoptry envelope; secretion

DOI:  https://doi.org/10.1080/21505594.2026.2707806
Biomolecules. 2026 Jul 03. pii: 978. [Epub ahead of print]16(7):

Design, Synthesis, and Antitumor Activities of Novel Coumarin-Based Histone Deacetylase Inhibitors.

Sichang Yan, Jie Chang, Dongyu Lei, Xiangyang Lv, Yanzhuo Li, Yue Zhuo, Lu Jin, Le Pan.

  Histone deacetylases (HDACs) are important epigenetic regulatory enzymes contributing to cancer proliferation, which could be critical targets in cancer therapy. The structural similarities of the existing HDAC inhibitors have resulted in an increase in the drug resistance. In this study, coumarin was employed as the core scaffold for structural derivatisation to develop a novel class of HDAC inhibitors based on computer-aided design (CADD). Their anti-tumor activity was evaluated against esophageal squamous cell lines. The results showed that most compounds exhibited potent anti-proliferative activity against KYSE70 and KYSE150. Among them, compound 4s and 4p exhibited the most potent activity with IC50 values of 3.44 μM and 3.39 μM against KYSE70. To validate the target of the synthesized compounds, transcriptome sequencing was performed and the results revealed that a total of 487 genes were differentially expressed, including 190 up-regulated and 297 down-regulated genes. Among these, 79 genes were associated with the HDAC regulatory network, accounting for 16.2% of the differentially expressed genes. Molecular docking demonstrated that compound 4s could effectively enter the active site of HDAC, engaging with the cap group, zinc-binding group, and linker region. This multiple interaction network provides a structural basis for the potent inhibitory activity of compound 4s. In conclusion, a series of novel HDAC inhibitors with a coumarin scaffold were discovered, and their mode of action was revealed. This provides a valuable guide for the development of novel HDAC-targeting therapeutics.

Keywords:  HDAC inhibitors; anti-tumor activity; coumarin derivatives; design and synthesis; transcriptome sequencing

DOI:  https://doi.org/10.3390/biom16070978
Front Cell Infect Microbiol. 2026 ;16 1843632

The role of autophagy in protozoan infections.

Xinlei Wang, Chao Zhang, Jie Chen, Yufen Jin, Yanan Zhang, Jingtong Zheng.

  Autophagy is a conserved, lysosome-dependent degradation system that is used by a wide variety of eukaryotes; during autophagy, intracellular substances are transported to lysosomes for degradation, and thus, autophagy plays a crucial role in cell survival under stress conditions such as starvation and hypoxia. Additionally, cells can eliminate foreign pathogens, such as parasites, bacteria, and viruses, via autophagic clearance. Protozoa are categorized as either intracellular parasitic protozoa or extracellular parasites, which are mostly zoonotic pathogens that pose significant threats to public health. In recent years, research on the mutual influences of autophagy and protozoa has focused mainly on Toxoplasma and Plasmodium, with less focus on extracellular parasitic protozoa. In this review, we discuss the crucial role of autophagy in maintaining the dynamic equilibrium between host elimination of extracellular and intracellular parasites and parasitic exploitation of the host.

Keywords:  autophagy; host-pathogen interaction; protozoa; protozoan infections; zoonosis

DOI:  https://doi.org/10.3389/fcimb.2026.1843632
Pathogens. 2026 Jul 07. pii: 718. [Epub ahead of print]15(7):

Acetylome Analysis in Vibrio vulnificus MO6-24/O Reveals Extensive Lysine Acetylation in Carbon Metabolism and Protein Synthesis Pathways: A Pilot Study.

Yurong Song, Ming Cheng, Xiaoling Li, Ying Wang, Yulong Zong, Jing Li, Guozhong Chen, Shiyong Chen, Yuan Cao.

  Vibrio vulnificus (V. vulnificus) is a type of bacterium commonly found in estuarine environments. It can cause necrotizing wound infections and sepsis, both of which are associated with high mortality rates. Protein lysine acetylation is a widespread post-translational modification (PTM) of proteins, and it participates in numerous cellular processes, including regulation, in bacteria. However, the finer landscape of lysine acetylation in V. vulnificus remains unexplored. In this study, acetylated proteins with low cellular abundance were enriched from V. vulnificus MO6-24/O using anti-acetyl-lysine immunoprecipitation and identified using LC-MS/MS, and the acetylation was further confirmed by Western blot analysis. We mapped 2035 lysine acetylation sites to 841 proteins, accounting for approximately 18.5% of the entire protein sequence of V. vulnificus MO6-24/O. Comprehensive bioinformatic characterization of the acetylome indicated that lysine acetylation is associated with metabolic regulation, particularly targeting enzymes regulating carbon metabolic functions and biosynthesis. In addition, sequence motif analysis identified two conserved patterns surrounding acetylated lysines: enrichment of lysine or arginine residues at the +4/+5 positions, and a preference for tyrosine, histidine, or phenylalanine residues at the -1/+1 positions. Furthermore, analysis of the protein-protein interaction network indicated that lysine acetylation influences numerous molecular interactions among proteins. Collectively, this acetylome investigation establishes a foundation for future studies aimed at elucidating physiological functions of protein lysine acetylation in V. vulnificus.

Keywords:  Vibrio vulnificus; acetylome; carbon metabolism; interaction network; lysine acetylation; lysine acetylation motif

DOI:  https://doi.org/10.3390/pathogens15070718
Cells. 2026 Jul 14. pii: 1267. [Epub ahead of print]15(14):

Low-Glucose Metabolic Remodeling Drives Reversible Sequestration of Yeast S-Adenosylmethionine (AdoMet) Synthase.

Hasnat Ahmed, Ahmad Sleiman, Mohamad Rahal, Eyas Hanbali, Marwa Aboumourad, Daniel Adebayo, Eseiwi Obaseki, Lindsey Kreinbring, Scott Miller, Elias Ayoub, Allison Jacobson, Andrew Rassam, Xun Bao, Jing Li, Hanaa Hariri.

  S-adenosylmethionine (AdoMet) is a central metabolite required for methylation and sulfur metabolism, synthesized in Saccharomyces cerevisiae by the cytosolic enzymes Sam1 and Sam2. While transcriptional and metabolic regulation of AdoMet biosynthesis have been extensively studied, how metabolic state influences the spatial organization of AdoMet synthases remains incompletely understood. Herein, we show that Sam1 undergoes robust and reversible relocalization into cytosolic assemblies under low-glucose metabolic states. These assemblies are dynamic and dissolve rapidly upon glucose refeeding in a manner that requires metabolizable glucose, indicating regulation by metabolic flux. While Sam1 assemblies associate with stress granules during prolonged starvation, a substantial fraction form independently under acute low-glucose conditions, indicating that Sam1 relocalization is not fully coordinated with canonical stress granule formation. Untargeted metabolomic analysis indicates that AdoMet pools are broadly maintained, while a targeted fluorescence-based enzymatic assay detects a modest but reproducible reduction in AdoMet under low-glucose conditions. Together, these findings define a metabolically gated mechanism for reversible Sam1 sequestration and support a model in which spatial organization of metabolic enzymes is coupled to cellular metabolic state.

Keywords:  S-adenosylmethionine (AdoMet); enzyme assemblies; metabolic flux; nutrient stress

DOI:  https://doi.org/10.3390/cells15141267
PLoS Comput Biol. 2026 Jul;22(7): e1014557

Deciphering chromatin architecture and dynamics in Plasmodium falciparum using the nucDetective pipeline.

Simon Holzinger, Leo Schmutterer, Victoria Marie Rothe, Maria Theresia Watzlowik, Uwe Schwartz, Gernot Längst.

High-resolution analysis of cellular chromatin structure is crucial for uncovering developmental and cell-type-specific regulatory networks. We developed the nucDetective pipeline to provide a comprehensive evaluation of chromatin organisation. This involves assessing nucleosome positioning, occupancy, fuzziness, and array regularity. The pipeline was benchmarked by analysing the chromatin structure of the malaria-causing parasite Plasmodium falciparum (Pf) during its erythrocytic development cycle. Pf is characterised by a unique chromatin landscape, exhibiting unstable nucleosomes and a genomic AT-content exceeding 80%, which presents challenges for standard MNase-seq analysis of chromatin. The nucDetective pipeline provides specific, high-resolution nucleosome profiles for the different asexual stages of Pf, monitoring the dynamics of individual nucleosomes. Contrary to the current view of irregular chromatin, we demonstrate for the first time regular phased nucleosome arrays downstream of TSSs, which, together with the established +1 nucleosome and upstream nucleosome-depleted region, reveal a complete canonical eukaryotic promoter architecture in Pf. The global mean nucleosome repeat length varies from 176 bp to 185 bp depending on the developmental stage. Stage specific changes in nucleosome positioning occur locally in intergenic regulatory regions, which are characterized by specific histone modifications and variants. Dynamic nucleosomes correlate with DNA accessibility, gene expression and determine the access to transcription factor binding sites in Pf. The highly regular chromatin structure, with stage-specific structural alterations, emphasises the important role of epigenetic mechanisms in regulating the complex life cycles of Pf.

DOI: https://doi.org/10.1371/journal.pcbi.1014557
Biochim Biophys Acta Rev Cancer. 2026 Jul 30. pii: S0304-419X(26)00143-5. [Epub ahead of print] 189671

Temporal and spatial molecular heterogeneity of gastric cancer: Roles of post-translational modifications.

Tian-Ao Han, Yuan Zhang, Ren-Chao Zhang, Chen Huang.

  Gastric cancer (GC) is a multifactorial disease responsible for significant global mortality, with approximately 1 million deaths annually. Its progression is governed by diverse molecular mechanisms, particularly protein post-translational modifications (PTMs), which play pivotal roles in either promoting or suppressing tumorigenesis. With the advent of spatiotemporal molecular omics, increasing emphasis has been placed on elucidating the temporal and spatial heterogeneity of GC. In this review, we systematically outline the temporal heterogeneity of GC across different developmental stages, integrating underlying mechanisms such as PTMs, including lactylation, ubiquitination, SUMOylation, which are the most associated ones influencing GC's spatiotemporal heterogeneity. Furthermore, we delineate the spatial heterogeneity of GC through comparative analyses of distinct features, such as proximal versus distal GC, primary versus metastatic lesions, superficial versus deep regions within primary tumors, and gastric mucus-secreting versus non-mucus-secreting adenocarcinoma. By synthesizing key molecular mechanisms and regulatory factors, this review aims to provide valuable insights for clinical practice, facilitating the advancement of precision medicine strategies-biomarker-guided therapy selection and spatial profiling for surgery planning, etc.

Keywords:  Gastric cancer; Molecular medicine; Post-translational modifications; Precision medicine; Spatiotemporal analysis; Translational medicine

DOI:  https://doi.org/10.1016/j.bbcan.2026.189671
Chembiochem. 2026 Jul 29. 27(14): e70473

Label-Free Monitoring of Cancer-Associated Fibroblast Activation Using NAD(P)H Fluorescence Lifetime Imaging.

Nina Anseeuw, Carmen Escalona-Noguero, Giel Vankevelaer, Samet Aytekin, Olivier De Wever, Ilaria Elia, Susana Rocha, Guillermo Solís-Fernández.

  Cancer-associated fibroblasts (CAFs) are key regulators of tumor progression, yet their activation state is commonly assessed using static, endpoint assays that do not allow dynamic analysis in living cells. Although CAF activation is accompanied by pronounced metabolic remodeling, label-free approaches that exploit these changes for real-time monitoring remain limited. Here, we demonstrate that NAD(P)H fluorescence lifetime imaging microscopy (FLIM) can be used to monitor activation-associated metabolic remodeling. CAFs activated with transforming growth factor beta (TGF-β) exhibit a reproducible shift toward longer NAD(P)H fluorescence lifetimes compared to non-activated cells, consistent with changes in the relative contributions of free and protein-bound NAD(P)H. By combining live-cell FLIM with α-smooth muscle actin staining in the same cells, we directly link metabolic signatures to cellular activation state. We further demonstrate the potential of this approach to dynamically monitor CAF activation in live, migrating cells. Together, these results establish NAD(P)H fluorescence lifetime imaging as a label-free metabolic approach for monitoring CAF activation dynamics, complementing conventional marker-based methods and enabling continuous monitoring of tumor-stroma interactions.

Keywords:  Cancer‐Associated Fibroblasts; Cellular metabolism; Fluorescence‐Lifetime Imaging Microscopy; Label free biosensing; Metabolic imaging; NADH fluorescence; TGF‐β signalling

DOI:  https://doi.org/10.1002/cbic.70473