bims-matara 2026-11-01 papers

on MTOR

Issue of 2026–11–01
eighteen papers selected by
Lucas van Endert, Institut du Cerveau and Alisa Kirkin, Université de Fribourg

Rethinking kidney cancer through the lens of microphthalmia (MiT)/transcription factor E (TFE) Biology.
mTOR and Extracellular Vesicles: Linking Chemical Stress to Ecological Communication.
mTOR Inhibition Strategies in Oropharyngeal Carcinoma.
P4HA3 promotes gastric cancer cell proliferation and invasion via the PI3K/AKT/mTOR signaling pathway and regulates glycolytic metabolic reprogramming.
STRADA deficiency impairs cortical interneuron development in humans and mice.
RapC Is Involved in the Development of Dictyostelium Through the Regulation of Autophagy and Calcium Signaling.
AMPK activation stabilizes HIF-2α by disrupting VHL binding and promotes erythroid differentiation under anemic stress via the mTOR pathway.
Relaxin-2 protects synaptic integrity via hippocampal NLRP3-mTOR-autophagy pathways in an age-dependent Alzheimer-like model.
Rapamycin attenuates bone cancer pain and tumor progression associated with reduced NE/NETosis-related molecular signatures in a mouse model.
KDM3B regulates hair follicle stem cell differentiation by activating the mTOR-dependent translation program.
Targeting PI3K/AKT/mTOR Pathway Exerts Inhibitory Effects on Endothelial Cells from Generalized Lymphatic Anomaly.
Dental Pulp Stem Cell-Derived Small Extracellular Vesicle miR-100-5p Attenuates Neuronal Oxidative Stress via Regulating the mTOR-Autophagy Signaling Axis.
Hydrogel bead-based 3D multiplex platform for high precision and high throughput quantification of mTORC1-4E-BP1/2-dependent translation.
NLRC4 Acts as a Non-Canonical Tumour Suppressor in Lung Adenocarcinoma by Inhibiting PI3K/AKT/mTOR Signalling to Induce Protective Autophagy.
miRNA-543 attenuates placental angiogenesis in female fetal growth restriction by targeting ANGPT2 and inhibiting the PI3K/AKT/mTOR pathway.
From light perception to energy signaling: integrating carbon dynamics with the Tre6P-SnRK1-TOR growth-stress axis.
Metformin (DMBG) ameliorates diabetic erectile dysfunction via the AMPK/mTOR pathway in association with endothelial autophagy reactivation.
Esculetin alleviates mouse transplant rejection by modulating CD4+ T cell functional responses and glycolytic metabolism via the PI3K-AKT-mTOR axis.

Trends Mol Med. 2026 Oct 26. pii: S1471-4914(26)00227-3. [Epub ahead of print]

Rethinking kidney cancer through the lens of microphthalmia (MiT)/transcription factor E (TFE) Biology.

Chiara Di Malta.

  Microphthalmia/transcription factor E (MiT/TFE) transcription factors are central regulators of lysosomal function, mechanistic target of rapamycin complex 1 (mTORC1) signaling, and cellular metabolism, and their dysregulation is increasingly recognized as a key driver of kidney cancer. This forum provides an overview of the molecular mechanisms underlying MiT/TFE-associated renal tumorigenesis and discusses their implications for cancer biology and therapy.

Keywords:  FLCN–FNIP; TFE3; TFEB; kidney cancer

DOI:  https://doi.org/10.1016/j.molmed.2026.09.011
Chimia (Aarau). 2026 Oct 28. 80(10): 685-694

mTOR and Extracellular Vesicles: Linking Chemical Stress to Ecological Communication.

Ksenia J Groh.

  Environmental stress, including that exerted by exposure to toxic chemicals, can shift cellular priorities from growth towards maintenance and communication. These changes in cellular resource allocation are coordinated by integrative molecular hubs such as the mechanistic Target of Rapamycin (mTOR) pathway. It regulates both the autophagy, which supports maintenance, and the release of extracellular vesicles (EVs), which mediate intercellular communication. This review discusses these interconnected processes and argues that environmental contaminants may influence not only intracellular signaling but also the molecular information exchanged among cells and potentially among organisms. EV-focused studies, performed at the intersection of chemical ecology, molecular biology, and ecotoxicology, can advance mechanistic understanding and open new opportunities for non-invasive environmental biomonitoring.

Keywords:  Chemical pollution; Environmental monitoring; Extracellular vesicle; Mechanistic Target of Rapamycin (mTOR)

DOI:  https://doi.org/10.2533/chimia.2026.685
Anticancer Res. 2026 Nov;46(11): 5947-5954

mTOR Inhibition Strategies in Oropharyngeal Carcinoma.

Georgios Papanastasiou, Aristeidis Chrysovergis, Vasileios Papanikolaou, Despoina Spyropoulou, Maria Adamopoulou, Ioanna E Metaxas, Sotirios Papouliakos, Evangelos Tsiambas, Vasileios Ragos.

  Oropharyngeal carcinoma (OPHC) belongs to the head and neck carcinoma (HNC) superfamily of malignant entities. Squamous cell carcinoma (SCC) represents the main histological type in them. Concerning the molecular substrate of OPHSCCs, imbalances in signal transduction pathways are implicated in their onset and progression. Among them, the phosphatidylinositol 3-kinase (PI3K)/tumor suppressor phosphatase and tensin homologue deleted on chromosome ten (PTEN)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) pathway deregulation critically affects the clinico-pathological features and biological behavior of the corresponding malignancies. Especially, the mTOR oncogene (gene locus: 1p36.2) overactivation, due to point mutations and gene amplification, is a relatively frequent genetic event in OPHSCCs. For this reason, the molecule is a target for specific oncological therapeutic regimens. There is an increasing interest in developing and applying novel mTOR inhibitors in subsets of patients with the malignancy that demonstrate specific genetic signatures. In the current review, we focused on the mTOR gene and protein characteristics, their deregulation mechanisms in OPHSCCs and the conventional and novel agents that target and disrupt mTOR oncogenic activity in them.

Keywords:  carcinoma; mTOR; oncogene; oral; pharynx; review; signaling pathway

DOI:  https://doi.org/10.21873/anticanres.18430
Pak J Pharm Sci. 2027 Jan 01. 40(1): 313-325

P4HA3 promotes gastric cancer cell proliferation and invasion via the PI3K/AKT/mTOR signaling pathway and regulates glycolytic metabolic reprogramming.

Tao Wang, Xiao Xu, Chen Jin, Yubing Zhong.

   BACKGROUND: Dysregulated expression of prolyl 4-hydroxylase subunit alpha 3 (P4HA3) has been identified in Colon cancer, breast cancer, head and neck cancers, etc. and is closely associated with tumor progression. However, its specific roles in gastric cancer (GC), particularly regarding alterations in metabolic pathways, are remain unknown.
OBJECTIVES: This study investigated how P4HA3 influences glycolytic metabolic reprogramming and the proliferation and invasion of gastric cancer cells via the PI3K/AKT/mTOR signaling pathway.
METHODS: To determine P4HA3 levels in GC tissues and corresponding cell lines, bioinformatics methods, immunohistochemical (IHC) staining and Western blotting (WB) were conducted. Furthermore, gain- and loss-of-function experiments, including cell viability assays (CCK-8), colony-forming tests, scratch wound assays and Transwell invasion experiments, were utilized to explore the influence of P4HA3 on GC cell growth, motility and invasive capacity. Changes in cellular metabolism were assessed using the Seahorse system (under normoxic conditions), including measurements of basal metabolism, glycolytic capacity and mitochondrial respiration (oxidative phosphorylation), as well as Western blot analysis of key glycolytic proteins. Additionally, the involvement of the PI3K/AKT/mTOR signaling axis was examined by gene set enrichment analysis (GSEA), WB analysis and pharmacological inhibition studies.
RESULTS: GC tissues and cell lines exhibited markedly increased levels of P4HA3, correlating significantly with decreased overall survival (OS). Experimental data indicated that P4HA3 facilitated GC cell proliferation, migration and invasion. In addition, P4HA3 enhanced glycolytic activity and increased the expression of key glycolysis-related proteins. Unexpectedly, P4HA3 also increased mitochondrial respiration under the assay conditions, suggesting that P4HA3 may induce a broader metabolic activation rather than an exclusively glycolytic shift. Mechanistically, the glycolysis-enhancing effects of P4HA3 were found to require PI3K/AKT/mTOR signaling (as shown by inhibitor studies).
CONCLUSION: P4HA3 promotes glycolysis via the PI3K/AKT/mTOR signaling pathway, thereby promoting the progression of gastric cancer.

Keywords:  Gastric cancer; Glycolytic reprogramming; Invasion; P4HA3; PI3K/AKT/mTOR signaling pathway; Tumor proliferation

DOI:  https://doi.org/10.36721/PJPS/2026.40.16437.1
Neurobiol Dis. 2026 Oct 27. pii: S0969-9961(26)00411-0. [Epub ahead of print]231 107665

STRADA deficiency impairs cortical interneuron development in humans and mice.

Ria K Parikh, Asmaa Hijazi, ThachVu H Nguyen, Meghna Pandey, Reana Young-Morrison, Danya A Adams, Sandesh Kamdi, Sophie Tran, Vincent J Carson, Philip H Iffland, Louis T Dang, Peter B Crino, Whitney E Parker.

  Polyhydramnios, Megalencephaly, and Symptomatic Epilepsy syndrome (PMSE/STRADA-related disease) is a rare neurodevelopmental disorder characterized by megalencephaly, early-onset drug-resistant epilepsy, neurocognitive impairment, and high early mortality, often due to status epilepticus. PMSE is caused by a multiexon deletion in the STRADA/LYK5 gene, encoding STRADA protein, which regulates the mechanistic target of rapamycin (mTOR) pathway. GABAergic inhibitory interneurons (INs) critically modulate excitatory:inhibitory balance in cortical networks, and IN deficits contribute to epileptogenesis in several epileptic encephalopathies. However, no studies to date have investigated INs in PMSE. We demonstrate that Strada/STRADA loss causes a reduction of multiple IN subtypes in the cerebral cortex and a corresponding increase in the striatum, representative of the remnant ganglionic eminence progenitor origin, in both Strada-/- mice and in a case of human PMSE brain tissue, suggestive of an IN migration defect. RNA sequencing comparing wildtype control to Strada-/- mouse cortex and striatum corroborates these findings, revealing increased IN-related gene expression (e.g., Dlx2) in the striatum. mTOR-associated genes (Lamp5, Cryab) are differentially expressed in the developing cortex, and cytoskeletal-associated genes (e.g., Tppp3, Kank4, Map1a) are differentially expressed in the developing cortex and mature striatum of Strada-/- mice. Functional validation confirms enlarged INs in Strada/STRADA-deficient brain, associated with mTOR hyperactivation. Together, these results suggest that STRADA/Strada loss may contribute to failed IN migration, the first such report in a developmental mTORassociated megalencephaly syndrome, highlighting INs as a therapeutic target for seizure prevention in PMSE.

Keywords:  Corticogenesis; Developmental epileptic encephalopathy; Megalencephaly; PMSE; mTOR

DOI:  https://doi.org/10.1016/j.nbd.2026.107665
Cells. 2026 Oct 15. pii: 1878. [Epub ahead of print]15(20):

RapC Is Involved in the Development of Dictyostelium Through the Regulation of Autophagy and Calcium Signaling.

Nara Han, Uri Han, Dahyeon Kim, Wonbum Kim, Taeck Joong Jeon.

  RapC is a Rap-subfamily small GTPase in Dictyostelium whose developmental function remains largely uncharacterized. Upon starvation, individual amoebae aggregate and undergo morphogenetic transitions to ultimately produce a fruiting body, a process requiring precise spatiotemporal regulation of a single apical tip as the primary organizing center. Here, we report that RapC is required for tip organizer formation during multicellular development by modulating autophagy and intracellular calcium signaling. rapC null cells develop normally through aggregation but consistently form three to four apical tips per aggregate rather than the single organizer tip characteristic of wild-type cells. Pharmacological phenocopy experiments revealed that caffeine and the selective TORC1 inhibitor rapamycin recapitulate the multi-tip phenotype, whereas the PI3K inhibitor LY294002 and the pan-TOR inhibitor Torin2 do not, implicating TORC1 signaling as the relevant downstream effector. In line with this, rapC null cells exhibit normal GFP-PH:CRAC translocation kinetics and slightly elevated PKB/PKBR1 phosphorylation, ruling out defects in the PI3K-TORC2-PKB signaling axis. Instead, rapC null cells display markedly elevated autophagic flux as assessed by the tandem RFP-GFP-Atg8 reporter assay, consistent with attenuated TORC1-mediated autophagy suppression. Furthermore, aggregation-competent rapC null cells show increased intracellular calcium ([Ca2+]i) upon external calcium stimulation, a response absent in wild-type cells and not reproduced by rapamycin, indicating a TORC1-independent role for RapC in calcium homeostasis. Together, our findings establish that RapC promotes normal Dictyostelium development by supporting TORC1 activity to suppress excessive autophagic flux and by maintaining [Ca2+]i homeostasis through a TORC1-independent mechanism.

Keywords:  Dictyostelium; Rap GTPase; RapC; TORC1; autophagy

DOI:  https://doi.org/10.3390/cells15201878
Front Physiol. 2026 ;17 1932780

AMPK activation stabilizes HIF-2α by disrupting VHL binding and promotes erythroid differentiation under anemic stress via the mTOR pathway.

Yun Cen, Pingjing Cai, Jinjin Jiang, Lihua Huang, Yiwei Hua.

   Objective: Anemia affects over two billion people globally, and ESA resistance poses major clinical hurdles. HIF-2α governs erythropoiesis, yet how AMPK/mTOR energy sensors regulate HIF-2α in erythroid cells remains unclear. This study explored whether AMPK activation stabilizes HIF-2α via impaired VHL binding to boost erythroid differentiation under anemic stress through mTOR signaling.
Methods: UT-7/EPO cells were cultured under 1% O2 with low EPO to mimic anemic stress, treated with AMPK/mTOR pharmacological modulators. CHX chase and Co-IP detected HIF-2α stability and VHL interaction; Ser672 phosphorylation, erythroid and iron-related genes, hepcidin and intracellular iron were measured.
Results: Anemic stress suppressed cell viability, stabilized HIF-2α, activated AMPK and inhibited mTOR, upregulated erythroid markers and triggered apoptosis. AICAR extended HIF-2α half-life, disrupted HIF-2α-VHL binding and elevated Ser672 phosphorylation, while mTOR activator MHY1485 reversed these effects. AMPK activation elevated EPOR, DMT1, FPN1, lowered hepcidin and ferritin, and restored intracellular iron; these effects vanished upon HIF-2α knockdown or mTOR overactivation.
Conclusions: AMPK stabilizes HIF-2α by blocking VHL binding to facilitate erythroid maturation and iron homeostasis under anemic stress via mTOR. The AMPK/mTOR/HIF-2α cascade controls stress erythropoiesis, and targeting this axis may treat ESA-insensitive anemia.

Keywords:  AMPK; HIF-2α; VHL; anemia; erythroid differentiation; mTOR

DOI:  https://doi.org/10.3389/fphys.2026.1932780
Acta Pharm. 2026 Oct 26.

Relaxin-2 protects synaptic integrity via hippocampal NLRP3-mTOR-autophagy pathways in an age-dependent Alzheimer-like model.

Daohe Wang, Zhigang Zhao, Yumei Ding.

  Alzheimer disease (AD) is associated with age-related synaptic degeneration, neuroinflammation, and impaired autophagy. The present study aimed to investigate the age-dependent neuroprotective effects of relaxin-2 (RLN2) in amyloid-beta 1-42 (Aβ1 - 42)-induced AD-like pathology and hypothesised that RLN2 would provide greater therapeutic benefit in young than aged mice. Young (3-4 months) and aged (12-14 months) mice were intracerebrally (i.c.) injected with amyloid-β (Aβ1 - 42) to develop an AD-like phenotype and treated with RLN2 or a control. Western blot analysis of hippocampal tissues was performed to assess the expression levels of ionised calcium-binding adaptor molecule 1 (Iba1) (microglial activation marker), NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome components, and mechanistic target of rapamycin (mTOR) pathway activity. Neuroinflammation was determined by interleukin-1 beta (IL-1β) and tumour necrosis factor-alpha (TNF-α), while synaptic integrity was assessed by synaptophysin and PSD-95. The Morris Water Maze (MWM) and Novel Object Recognition (NOR) tests were used to measure cognitive and memory performance. Neuroinflammation markers IL-1β and TNF-α were significantly elevated in Aβ1 - 42-injected mice, with aged mice showing a greater increase than young mice. RLN2 treatment markedly reduced IL-1β and TNF-α levels in young mice, whereas the reduction was limited in aged mice. RLN2 treatment had a significant inhibitory effect on hippocampal NLRP3 signalling, reduced mTOR signalling, modulated autophagy-related protein expression, preserved synaptic proteins, and improved cognitive and memory performance (decrease in the MWM escape latency and increase in the NOR discrimination index). RLN2 was not effective in the aged mice, and there was persistent inflammatory activation, impaired autophagy, minimal improvement in synaptic recovery, and a moderate improvement in cognition. These findings suggest that RLN2 could be a potential therapeutic candidate for early-stage AD by targeting neuroinflammation, mTOR-mediated autophagy-related dysregulation, and synaptic loss, while also potentially providing a basis for age-based stratification of treatment responsiveness; however, further studies are required to determine its utility as a biomarker of treatment response or disease progression. Overall, RLN2 exerted neuro-protective effects in i.c. Aβ1 - 42-induced AD-like pathology in an age-dependent manner.

Keywords:  NLRP3 inflammasome; autophagy; microglia; relaxin-2; synaptic integrity

DOI:  https://doi.org/10.2478/acph-2026-0023
Int Immunopharmacol. 2026 Oct 28. pii: S1567-5769(26)01458-X. [Epub ahead of print]191(Pt 1): 117610

Rapamycin attenuates bone cancer pain and tumor progression associated with reduced NE/NETosis-related molecular signatures in a mouse model.

Wen Li, Xiaochen Wang, Long Wang, Guo Tang, Yang Li, Zeguo Feng.

  Bone cancer pain (BCP) is a refractory condition associated with tumor metastasis for which effective treatment options remain limited. Although NETosis-related inflammatory processes and neutrophil elastase (NE) have been implicated in pathological pain, their association with BCP remains insufficiently characterized. Here, we investigated NETosis-related molecular alterations in a BCP mouse model and evaluated the analgesic and antitumor effects of rapamycin, a mammalian target of rapamycin (mTOR) inhibitor with immunomodulatory properties. The BCP model was established by injecting Lewis lung carcinoma cells into the femur, resulting in progressive mechanical allodynia, heat hyperalgesia, and spontaneous pain. Spinal transcriptomic analysis revealed enrichment of genes associated with NETosis-related processes, with upregulated expression of Elane and Mpo mRNA. Exploratory molecular docking was performed to visualize a possible interaction pattern between rapamycin and NE. In vivo, rapamycin (1.5-5 mg/kg, i.p.) produced dose-related attenuation of pain-related behaviors. The 5 mg/kg dose showed the greatest behavioral improvement and was selected for subsequent analyses. Western blotting and immunofluorescence demonstrated that rapamycin treatment was associated with reduced levels of NETosis-associated markers, including NE, MPO, and citrullinated histone H3, in the spinal cord. Rapamycin treatment was also associated with reduced local tumor growth. Collectively, these findings suggest that NE/NETosis-related molecular alterations may represent a relevant feature of BCP pathophysiology. Rapamycin attenuated pain-related behaviors and was associated with reduced local tumor burden and lower levels of NE/NETosis-related molecular markers in this mouse model, suggesting that these molecular alterations may accompany the observed beneficial effects.

Keywords:  Bone cancer pain; Hyperalgesia; Neutrophil elastase; Neutrophil extracellular trap formation; Rapamycin

DOI:  https://doi.org/10.1016/j.intimp.2026.117610
J Mol Cell Biol. 2026 Oct 30. pii: mjag046. [Epub ahead of print]

KDM3B regulates hair follicle stem cell differentiation by activating the mTOR-dependent translation program.

Chaofan Lv, Hao Wang, Fan Meng, Meijun Zhou, Jiaqi Fan, Runsheng Chen, Jian Zhang, Haisheng Yu, Yan Qin.

  Hair follicle (HF) development and regeneration rely on the precise differentiation of hair follicle stem cells (HFSCs), a process governed by epigenetic regulation. While nutrient-sensing mTOR signaling is known to be pivotal, the upstream epigenetic triggers and the downstream events orchestrating this pathway remain poorly defined. The histone H3K9me2 demethylase KDM3B has recently emerged as a key player in stem cell fate decisions. By integrating recent high-throughput sequencing data and employing a conditional knockout mouse model, we demonstrated that KDM3B was essential for HFSC differentiation, as its loss led to severe defects in postnatal HF development and HFSC differentiation. Mechanistically, we identified the mTOR signaling pathway and its downstream protein translation machinery as the primary effectors of KDM3B. KDM3B activated mTOR complex 1 (mTORC1) signaling, which subsequently upregulated the TOP mRNA-binding protein LARP1 to drive ribosome biogenesis and global protein synthesis. Our work delineated a complete 'KDM3B-mTORC1-LARP1-ribosome' axis that functionally integrates epigenetic control with translational regulation to direct HFSC differentiation. These findings not only establish KDM3B as a novel key regulator in skin epithelia but also provide the first comprehensive mechanistic link from an epigenetic modifier through the mTOR pathway to the ribosomal translation program within the HF model.

Keywords:  AKT–mTOR pathway; KDM3B; hair follicle differentiation; hair follicle stem cells; ribosome

DOI:  https://doi.org/10.1093/jmcb/mjag046
Cells. 2026 Oct 13. pii: 1860. [Epub ahead of print]15(20):

Targeting PI3K/AKT/mTOR Pathway Exerts Inhibitory Effects on Endothelial Cells from Generalized Lymphatic Anomaly.

Bruno Lorusso, Laura Finato, Antonella Nogara, Giovanni Roti, Anna Montanaro, Gregorio Monica, Alice Andreani, Gallia Graiani, Pellegrino Crafa, Enrico Maria Silini, Letizia Gnetti, Francesco Paolo Pilato, Lorena Bottarelli, Cinzia Azzoni, Giuseppe Cerasoli, Federico Quaini, Costanza Anna Maria Lagrasta.

  Generalized lymphatic anomaly (GLA) is a congenital, non-neoplastic vascular anomaly characterized by diffuse or multifocal involvement and infiltration of multiple organs. Most GLA cases harbor "hotspot" somatic missense single nucleotide mutations in the PIK3CA gene, whereas a minority involve mutations in other genes. Recently, rapamycin and its homologs have shown encouraging results in the treatment of alterations affecting the PI3K/AKT/mTOR pathway. Therefore, the present work aimed to expand the immunophenotypic characterization of the lymphatic lesion at the tissue level and to evaluate the in vitro viability, migration, and angiogenic capacity of endothelial cells freshly isolated from a PIK3CA-mutated GLA sample, following treatment with alpelisib, rapamycin, dactolisib, and buparlisib. We detected the p.Gln546Lys amino acid substitution in the PI3Kca helical domain in GLA-derived endothelial cells. Then, by immunohistochemical analysis, we documented altered expression of mTOR and its well-characterized downstream substrates, which regulate growth, proliferation, migration, angiogenesis, and survival of ECs. Finally, pharmacological treatment reduced GLA-EC functional properties and down-modulated the hyperactivated PI3K downstream substrates. Overall, our results suggest that routine immunohistochemistry, integrated with phospho-protein analysis and next-generation sequencing, might improve histopathological classification of vascular anomalies and facilitate the development of personalized targeted therapies, particularly in recurrent or surgically challenging cases.

Keywords:  PI3Kca mutation; alpelisib; angiogenesis; buparlisib; dactolisib; generalized lymphatic anomaly; immunohistochemistry; lymphatic endothelial cells; migration; rapamycin

DOI:  https://doi.org/10.3390/cells15201860
Cells. 2026 Oct 11. pii: 1843. [Epub ahead of print]15(20):

Dental Pulp Stem Cell-Derived Small Extracellular Vesicle miR-100-5p Attenuates Neuronal Oxidative Stress via Regulating the mTOR-Autophagy Signaling Axis.

Wenli Zhou, Yanfang Zhao, Fengtang Yang, Wensheng Zhang, Xiuzhen Zhang.

  Excessive accumulation of reactive oxygen species (ROS) in the brain and the resulting oxidative stress are core drivers of neurodegenerative processes. Human dental pulp stem cells (DPSCs) secrete small extracellular vesicles (sEVs) abundant in functional microRNAs, which possess prominent regenerative capacity. The present study investigated the neuroprotective effects of DPSC-derived sEVs (DPSC-sEVs) against oxidative injury in SH-SY5Y neuronal cells and elucidated the underlying molecular mechanisms. DPSC identity was confirmed via flow cytometric immunophenotyping and multipotent differentiation assays. sEVs were isolated from DPSC-conditioned medium and characterized by transmission electron microscopy, nanoparticle tracking analysis and detection of canonical exosomal markers (CD63, CD81, and TSG101). An in vitro model of neuronal oxidative stress was established using hydrogen peroxide (H2O2). Cell viability, apoptosis, intracellular ROS levels, and lactate dehydrogenase (LDH) release, as well as the levels/activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA), were assessed to evaluate the protective effects of sEVs. Candidate miRNAs in sEVs were screened by quantitative real-time PCR (qPCR). Direct binding of miR-100-5p to the 3' untranslated region (3'UTR) of mammalian target of rapamycin (mTOR) was verified via dual-luciferase reporter assay. Autophagic activity was evaluated by immunoblotting and immunofluorescence staining of LC3B/p62. Pretreatment with DPSC-sEVs significantly ameliorated H2O2-induced neuronal oxidative damage and apoptosis. Mechanistically, sEV miR-100-5p conferred neuroprotective and antioxidant properties by inhibiting mTOR signaling and activating autophagy in vitro. Blockade of miR-100-5p or pharmacological inhibition of autophagy by 3-methyladenine (3-MA) largely abolished these protective effects. In conclusion, DPSC-sEVs attenuate neuronal oxidative stress by reversing mTOR-mediated suppression of autophagy, and this protective effect is dependent on the function of miR-100-5p in SH-SY5Y cells. The miR-100-5p/mTOR/autophagy axis mediates the neuroprotective effects of DPSC-sEVs, representing a promising therapeutic candidate for oxidative stress-associated neurodegenerative diseases that merits further in vivo validation.

Keywords:  autophagy; dental pulp stem cells; mTOR; microRNA; oxidative stress; small extracellular vesicles

DOI:  https://doi.org/10.3390/cells15201843
Anal Chim Acta. 2026 Dec 01. pii: S0003-2670(26)01215-8. [Epub ahead of print]1425 346265

Hydrogel bead-based 3D multiplex platform for high precision and high throughput quantification of mTORC1-4E-BP1/2-dependent translation.

Mariana Ferreira Pissarra, Lokman Alpsoy, Malte Lauritz Breyer, Vanessa Giossas, Kartik Jatwani, Karmel Sofia Gkika, Anthony Squire, Maithreyan Kuppusamy, Andrea Oeckinghaus, Martin Termathe, Jose Ramos Pittol, Christos G Gkogkas, Thomas Brandstetter, Jürgen Rühe, Ulrike Rehbein, Kathrin Thedieck.

   BACKGROUND: Quantifying protein phosphorylations and interactions across signaling networks is essential for quantitative biology and systems medicine. Current standard methods like immunoblotting offer limited throughput, precision, and scalability. We present an inverse-precision multiplex workflow based on a 3D hydrogel platform, enabling quantification of protein modifications and interactions within one assay. We chose the example of the mTORC1 (mTOR complex 1) kinase - 4E-BP1/2 signaling axis, clinically targeted by cancer therapies, which tunes protein biosynthesis to nutrient supply and stress.
RESULTS: We developed an inverse-precision design combining two orthogonal readouts, 4E-BP1/2 phosphorylation and eIF4E interaction. The design relies on two assays with inverse responses to mTORC1-dependent translation, offering inverse precision. Their combined analysis enhances the assay's robustness and dynamic range (LoD = 9% mTORC1 activity versus LoD = 15% for single readouts). Compared to conventional assays, 3D multiplexing increases precision by > fivefold (CV<4% versus 21% for immunoblotting around LoD), reduces sample consumption by 16-fold (7.8 μg protein input versus 125 μg in 2D format) and time effort by ≥ 80%, while offering a high degree of parallelization.
SIGNIFICANCE: Our workflow is compatible with small sample sizes, multiple technical and biological replicates, high-density dose-response curves and time-course data. We validated the assay for different physiological and experimental settings in cell culture and brain tissue. Standard microscopy equipment avoiding the need of specialized infrastructure, and assembly of custom antibody panels without dependence on fixed commercial kits present further advantages over established closed-panel systems. Its scalability, time-efficiency, and high precision make our platform a powerful tool for systems biology and precision medicine.

Keywords:  3D hydrogel platform; 4E-BP1; High throughput quantitation; Multiplexing; eIF4E; mTORC1

DOI:  https://doi.org/10.1016/j.aca.2026.346265
J Cell Mol Med. 2026 Oct;pii: e71420. [Epub ahead of print]30(20):

NLRC4 Acts as a Non-Canonical Tumour Suppressor in Lung Adenocarcinoma by Inhibiting PI3K/AKT/mTOR Signalling to Induce Protective Autophagy.

Yuxin Zhang, Shuangteng Liu, Rui Li, Mengyu Zhang, Guang Yang, Xinmiao Zhu, Yiqing Qu.

  Autophagy plays complex, context-dependent roles in lung adenocarcinoma (LUAD), but the upstream molecular drivers that govern this process remain elusive. NLRC4 is classically defined as a pyroptosis-inducing inflammasome sensor, yet its non-canonical, tumour-intrinsic function in LUAD progression remains largely unexplored. Through multi-omics screening and prognostic modelling across TCGA and GEO datasets, we identified NLRC4 as a critical autophagy-associated protective factor, which was validated in an institutional cohort of paired LUAD tissues. In vitro and in vivo assays, autophagic flux monitoring, reciprocal co-immunoprecipitation and genetic rescue experiments were employed to elucidate the underlying mechanism. NLRC4 protein was markedly downregulated in LUAD tissues alongside the tumour suppressor PTEN, correlating with advanced TNM stage and poor overall survival. Functionally, restoring NLRC4 expression inhibited LUAD cell proliferation, motility, invasion and epithelial-mesenchymal transition in vitro, whereas NLRC4 depletion accelerated xenograft tumour growth in vivo. Crucially, NLRC4 selectively stimulated complete autophagic flux without triggering Caspase-1/Gasdermin cleavage or lytic pyroptotic death. Mechanistically, NLRC4 physically interacted with NFKB1 (p105/p50) in a tumour-selective manner, relieving NF-κB-mediated transcriptional repression of PTEN. This restored PTEN expression, thereby tonically dampening the PI3K/AKT/mTOR signalling cascade. Genetic blockade of autophagy (siATG5) or PTEN ablation largely reversed the tumour-suppressive effects of NLRC4, confirming the functional dependency on the PTEN-autophagy axis. Our study establishes a non-canonical, tumour-intrinsic role for NLRC4 in restraining LUAD progression via the PI3K/AKT/mTOR axis to induce protective autophagy, positioning it as a promising prognostic biomarker and potential therapeutic target.

Keywords:  NLRC4; PI3K/AKT signalling; autophagy; lung adenocarcinoma; prognosis

DOI:  https://doi.org/10.1111/jcmm.71420
Front Cell Dev Biol. 2026 ;14 1912518

miRNA-543 attenuates placental angiogenesis in female fetal growth restriction by targeting ANGPT2 and inhibiting the PI3K/AKT/mTOR pathway.

Wenhui Song, Xuemei Yang, Chunyan Yang, Yuanyuan Zhao, Xinxian Zhang, Qing Guo.

   Background: Insufficient placental angiogenesis is a predominant pathological cause of fetal growth restriction (FGR). Placental capillary development is regulated in a sex-dependent manner, with female placentas exhibiting greater susceptibility to angiogenic dysfunction than male placentas. However, the sex-specific interplay between microRNAs (miRNAs) and human placental angiogenesis in FGR pathogenesis remains poorly defined. Emerging transcriptomic sequencing evidence has implicated miR-543 as a pivotal regulator of impaired angiogenesis in female FGR placentas. This study aimed to elucidate the sex-dependent mechanistic role of miR-543 in modulating placental angiogenesis in a cell-specific manner.
Method: FGR pregnancies were recruited, and placental tissues were collected after delivery. The dual-luciferase reporter assay was used to detect the direct binding site of miR-543 to ANGPT2. Next, immunohistochemistry (IHC) and immunofluorescence (IF) staining were used to detect the expression and localization of ANGPT2. To verify the cell-specific gene expression of miR-543 and ANGPT2, laser capture microdissection (LCM) was employed to isolate endothelial cells from human placentas. The gene expression levels of miR-543 and ANGPT2 were quantified by qRT-PCR. CCK8, wound healing, Transwell, and tube formation assays and flow cytometry were performed to assess the functional effects of miR-543 and ANGPT2 on the biological function of human umbilical vein endothelial cells (HUVECs). The PI3K/AKT/mTOR signaling pathway was predicted and verified by Western blotting.
Results: Significantly downregulated miR-543 expression was observed in the placentas of female FGR fetuses relative to matched normal controls. According to target gene prediction and the dual-luciferase reporter assay, ANGTP2 was identified as a target candidate of miR-543. Moreover, immunofluorescence staining and LCM analysis verified the cell-specific distribution of ANGPT2 in female FGR placentas, partly indicating its key involvement in placental capillary angiogenesis. Furthermore, dysregulated expression of miR-543 and ANGPT2 was closely correlated with aberrant proliferation, migration, and apoptosis of HUVECs. KEGG analysis revealed that the candidate targets of miR-543 were mainly enriched in the PI3K/AKT/mTOR pathway. Western blot analysis demonstrated that the PI3K/AKT/mTOR signaling pathway mediated the ANGPT2-induced attenuation of placental angiogenesis in female FGR.
Conclusion: miR-543 negatively regulated ANGPT2 expression, leading to impaired endothelial cell function via inactivation of the PI3K/AKT/mTOR signaling pathway. This, in turn, contributed to sex-specific inhibition of capillary angiogenesis in the placenta of female fetuses. These findings reveal a novel miRNA-mediated, sex-dependent regulatory mechanism underlying the pathogenesis of impaired placental angiogenesis.

Keywords:  angiopoietin 2; apoptosis; endothelial cell; fetal growth retardation; microRNA; placenta

DOI:  https://doi.org/10.3389/fcell.2026.1912518
Trends Biochem Sci. 2026 Oct 28. pii: S0968-0004(26)00317-8. [Epub ahead of print]

From light perception to energy signaling: integrating carbon dynamics with the Tre6P-SnRK1-TOR growth-stress axis.

Muhammad Asim, Zexian Zeng, Xiong Yan, Hongtao Liu.

  Plants growing in fluctuating environments require dynamic coordination between carbon availability and growth-stress decisions. Trehalose-6-phosphate (Tre6P), a sugar signal linked to sucrose status, helps coordinate these decisions by regulating SUCROSE NON-FERMENTING 1-RELATED KINASE 1 (SnRK1)- and TARGET OF RAPAMYCIN (TOR)-dependent growth programs. Here, we synthesize recent advances in photosynthetic carbon dynamics, carbon sensing, and Tre6P-SnRK1-TOR signaling, highlighting emerging regulatory layers including Class II TREHALOSE PHOSPHATE SYNTHASE proteins, SnRK1 compartmentalization, and stress-associated TOR regulation. We further discuss how different photosynthetic strategies of carbon fixation, including C3, C4, and crassulacean acid metabolism, as well as UV-B signaling through UV RESISTANCE LOCUS 8, influence carbon-energy signaling. Finally, we outline unresolved questions relevant to crop resilience under fluctuating environments.

Keywords:  Suc; Tre6P; carbon–energy homeostasis; carbon–energy signaling; light signaling; photosynthetic carbon fixation

DOI:  https://doi.org/10.1016/j.tibs.2026.09.012
J Reprod Immunol. 2026 Sep 20. pii: S0165-0378(26)00137-3. [Epub ahead of print]178 104968

Metformin (DMBG) ameliorates diabetic erectile dysfunction via the AMPK/mTOR pathway in association with endothelial autophagy reactivation.

Jiaxing Chen, Lingyun Hu, Yaqi Ding, Wangguan Zhou, Chaoyang Wang, Xuewu Zhou, Jianbo Hong.

   BACKGROUND: Diabetes mellitus erectile dysfunction (DMED) is a prevalent complication with limited treatment efficacy. Metformin (dimethylbiguanide, DMBG) shows therapeutic potential, but its underlying mechanisms remain unclear. This study investigated whether DMBG alleviates DMED in association with restored endothelial autophagy.
METHODS: A DMED rat model was induced by streptozotocin and validated by apomorphine screening. Rats were treated with DMBG or Sildenafil (positive control) for 30 days. Erectile function was assessed by intracavernosal pressure (ICP) and mean arterial pressure (MAP). Penile tissues were analyzed histologically and for autophagy markers. To decipher the cellular mechanism, primary cavernosal endothelial cells (PCECs) were isolated from each treatment group and subjected to functional assays including viability, angiogenesis, migration, and apoptosis. The role of the AMPK/mTOR pathway was interrogated through western blot and by using the AMPK inhibitor BAY-3827 in a rescue experiment.
RESULTS: DMBG significantly restored erectile function and cavernosal endothelial integrity, comparable to Sildenafil. It downregulated elevated serum VEGF-A and upregulated impaired penile autophagy markers (LC3-II, BECLIN1, ATG5). PCECs from DMBG-treated rats exhibited rescued viability, angiogenesis, migration, and reduced apoptosis, alongside reactivated autophagy. Mechanistically, DMBG activated AMPK and inhibited mTOR phosphorylation in PCECs, increasing eNOS. These protective effects were substantially attenuated by AMPK inhibition.
CONCLUSIONS: DMBG ameliorates DMED in association with improved endothelial function and reactivated autophagy. These beneficial effects are associated with activation of the AMPK/mTOR signaling pathway, highlighting a novel mechanistic basis for the use of DMBG in DMED management.

Keywords:  AMPK; Autophagy; Diabetic erectile dysfunction; Endothelial dysfunction; Metformin

DOI:  https://doi.org/10.1016/j.jri.2026.104968
Int Immunopharmacol. 2026 Oct 27. pii: S1567-5769(26)01421-9. [Epub ahead of print]191(Pt 1): 117573

Esculetin alleviates mouse transplant rejection by modulating CD4+ T cell functional responses and glycolytic metabolism via the PI3K-AKT-mTOR axis.

Tian Han, Biqiong Qu, Qingyao Chang, Rui Zhang.

   BACKGROUND: Immune rejection is a major cause of graft dysfunction following organ transplantation, and CD4+ T cells play a critical role in this process. Esculetin is a natural coumarin compound derived from the Chinese herbal medicine Fraxinus. Although its anti-tumor and anti-inflammatory effects have been extensively investigated, its potential role in transplant immunology remains unexplored.
METHODS: The effect of esculetin on transplant rejection and CD4+ T cells was evaluated by survival curve analysis, H&E staining, immunohistochemical staining, flow cytometry, and RNA sequencing. The effects of esculetin on the phosphoinositide 3-kinase (PI3K)-protein kinase B (AKT)-mechanistic target of rapamycin (mTOR) pathway, cell cycle, apoptosis, and glycolytic metabolism in CD4+ T cells were also examined. The functional rescue experiments were performed by combining esculetin with the PI3K activator 740Y-P and inhibitor LY294002.
RESULTS: Esculetin prolonged graft survival and attenuated graft injury. In addition, esculetin treatment suppressed CD4+ T cell functional responses. RNA sequencing of cultured CD4+ T cells revealed that genes downregulated by esculetin were enriched in the PI3K-AKT and cell cycle pathways. Mechanistically, esculetin inhibited PI3K-AKT-mTOR signaling, glycolysis, and cell cycle progression, while promoting apoptosis in CD4+ T cells. Furthermore, rescue experiments using PI3K modulators confirmed that esculetin acts through this pathway to suppress proliferation, induce apoptosis, and prolong graft survival.
CONCLUSIONS: The present study is the first to demonstrate that esculetin attenuates immune rejection in mouse skin and heart transplantation models by modulating the PI3K-AKT-mTOR signaling pathway in CD4+ T cells, thereby exerting inhibitory effects on proliferation, induction of apoptosis, and suppression of glycolytic metabolism.

Keywords:  CD4(+) T cells; Esculetin; Immune rejection; Organ transplant; PI3K-AKT-mTOR

DOI:  https://doi.org/10.1016/j.intimp.2026.117573