bims-auttor Biomed News
on Autophagy and mTOR
Issue of 2026–08–30
33 papers selected by
Viktor Korolchuk, Newcastle University



  1. Adv Sci (Weinh). 2026 Aug 27. e77428
      Intervertebral disc degeneration (IDD) is a leading cause of low back pain with incompletely understood mechanisms. Although autophagy dysfunction is a documented contributor to IDD, the precise pathobiological role of chaperone-mediated autophagy (CMA) remains poorly understood. Here, we demonstrate that CMA activity is downregulated in nucleus pulposus cells (NPCs) from IDD patients and IL-1β-induced rat intervertebral disc cell models, causing cytoplasmic accumulation of a novel CMA substrate, Midnolin (MIDN). Accumulated MIDN bypasses the ubiquitin-proteasome system and directly binds to Tuberous Sclerosis Complex 2 (TSC2), mediating its degradation. TSC2 loss relieves mechanistic target of rapamycin complex 1 (mTORC1) inhibition, resulting in mTORC1 hyperactivation, which drives cellular senescence, senescence-associated secretory phenotype (SASP), and extracellular matrix (ECM) degradation in NPCs. In vitro and in a rat caudal needle puncture model, MIDN knockdown (shRNA), CMA activation (LAMP2A overexpression), or mTORC1 inhibition (Rapamycin) significantly attenuated IL-1β or MIDN overexpression-induced senescence and disc degeneration. Our findings reveal an "Impaired CMA-MIDN accumulation-TSC2 degradation-mTORC1 activation" axis central to IDD pathogenesis, offering potential therapeutic targets.
    Keywords:  autophagy; cellular senescence; intervertebral disc degeneration; low back pain; proteostasis
    DOI:  https://doi.org/10.1002/advs.77428
  2. Nat Commun. 2026 Jul 22. pii: 8948. [Epub ahead of print]17(1):
      Autophagy intersects with endocytic trafficking to regulate extracellular vesicle (EV) biogenesis, but how upstream lipid-handling autophagy proteins influence this crosstalk is unclear. Here we show that the autophagy lipid-supply proteins ATG9A and ATG2A/B restrain small EV (sEV) secretion by promoting amphisome formation and controlling cellular lipid composition. Deletion of ATG9A or ATG2A/B in cells, which abolishes autophagosome biogenesis, causes a RAB27A-dependent increase in secretion of CD63-enriched, smaller sEVs, and accumulation of intraluminal vesicles within multivesicular endosomes. Under lysosomal inhibition, wild-type cells release LC3- and autophagy cargo receptor-positive sEVs, whereas ATG9A- and ATG2A/B-deficient cells, despite hypersecretion of sEVs, fail to load LC3 or canonical cargo receptors, indicating a block in amphisome-mediated export. Proteomics reveals selective depletion of autophagy receptors and ferritinophagy factors and enrichment of RNA-binding proteins and endosomal trafficking regulators in sEVs from ATG9A- and ATG2A/B-deficient cells. Whole-cell lipidomics uncovers extensive rewiring of the lipidome, with accumulation of ceramides and neutral lipids, altered phospholipid balance, and transcriptional remodeling of lipid metabolic enzymes, while neutral sphingomyelinase inhibition normalizes sEV output. These findings identify ATG9A and ATG2A/B as lipid-dependent gatekeepers that couple autophagosome and amphisome formation, regulating membrane partition between degradative autophagy and exosome-mediated secretion.
    DOI:  https://doi.org/10.1038/s41467-026-75742-x
  3. Autophagy. 2026 Aug 26.
      Mechanisms aimed at recovering from heat-induced damage are closely associated with the organism's ability to survive extreme temperature exposure. In such a scenario, we show that autophagy, as a cytoprotective mechanism, ensures recovery and viability after induced heat stress in Saccharomyces cerevisiae. Our findings indicate that heat shock triggers the targeted degradation of ubiquitinated protein aggregates, mediated by the macroaggrephagy receptor Cue5. Moreover, heat stress induces the turnover of the aggrephagy receptor Cct2 and the polyglutamine repeats of the HTT (huntingtin) protein (polyQ-HTT). Notably, even though Cct2 and polyQ-HTT degradation is vacuole-dependent, it is mediated autonomously of canonical autophagy pathways. Collectively, this study demonstrates a novel role of autophagy in maintaining protein homeostasis after heat stress in yeast and provides insights into the potential medical applications of heat treatment.
    Keywords:  Aggrephagy; Cct2; Cue5; autophagy; budding yeast; heat stress; polyQ-HTT
    DOI:  https://doi.org/10.1080/15548627.2026.2724473
  4. Autophagy. 2026 Aug 28.
      The identification of pathogenic autosomal recessive mutations in the gene encoding the PINK1 kinase provided early evidence linking mitochondrial dysfunction to neurodegeneration - in this case Parkinson's Disease. PINK1 has since become synonymous with mitophagy, with the prevailing model proposing two alternative fates. The first being partial import - inner-membrane penetration of its transmembrane domain (TMD) - followed by PARL-mediated cleavage and degradation. This happens in healthy mitochondria with a high membrane potential (ΔΨ) across the inner-membrane - required for passage of proteins into or across the inner-membrane. The second being surface stabilization, Parkin activation and initiation of mitophagy upon membrane depolarization. But what if PINK1 acts in active mitochondria as well? Our recent work identifies a third fate - matrix entry! The findings expand the biology of PINK1 beyond mitochondrial surveillance for quality control alone. They suggest an additional mitophagy-independent regulatory role within the matrix, which turns out to be governed by the unusual properties of its TMD for the conferral of a decisive conformational switch.
    Keywords:  Mitochondria; PARL; PINK1; Parkinson’s disease; transmembrane
    DOI:  https://doi.org/10.1080/15548627.2026.2726091
  5. J Inherit Metab Dis. 2026 Sep;49(5): e70245
      Lysosomal disorders (LDs) have traditionally been defined by intra-lysosomal substrate accumulation resulting from deficiencies of lysosomal enzymes or associated proteins. Advances in lysosomal biology have demonstrated that lysosomes function as central regulators of cellular signalling, membrane trafficking, autophagy, nutrient sensing, organelle communication and cellular homeostasis, expanding the spectrum of inherited disorders associated with lysosomal dysfunction beyond classical storage phenotypes. We developed a contemporary pathomechanistic nosology of inherited LDs through expert curation and targeted review of databases and published literature. Disorders were included when pathogenic variants resulted in lysosomal dysfunction as a major disease mechanism through defects affecting lysosomal degradation, membrane function, intracellular trafficking, biogenesis, autophagy-lysosome pathways or lysosome-related organelles. A total of 108 inherited lysosomal disorders caused by defects in 102 genes were identified and organised into 11 major disease categories. Neurologic and eye involvement were the most frequently affected organ-system categories, occurring in 80.6% and 68.5% of disorders, respectively. Digestive (including hepatosplenomegaly), dysmorphic, skeletal and haematological involvement occurred in 48.1%, 45.4%, 40.7% and 38.9% of disorders, respectively. Distinct phenotypic signatures were observed across disease categories despite substantial mechanistic overlap involving impaired autophagy, vesicular trafficking, lysosomal stress and altered organelle homeostasis. This proposed nosology extends disease classification beyond substrate accumulation alone and provides a biologically informed framework for disease classification, genomic interpretation, biomarker development, patient stratification and the development of mechanism-based therapies.
    Keywords:  autophagy; disease classification; endolysosomal pathway; inherited lysosomal disorders; inherited metabolic disorders; lysosomal storage disorders; lysosome; nosology
    DOI:  https://doi.org/10.1002/jimd.70245
  6. Nat Neurosci. 2026 Aug 24.
      Macroautophagy (autophagy) is a lysosome-dependent degradative pathway that encapsulates proteins and organelles within double-membraned vesicles and recycles the contents back into the cell. The interplay of autophagy with various membrane trafficking pathways is crucial for maintaining cellular homeostasis both under basal conditions and in response to environmental stress. In the CNS, neurons and glial cells rely on a spectrum of quality-control and recycling pathways, notably autophagy, to sustain the intricate functions of the brain. Genetic variants in autophagy genes are known to cause Mendelian disorders primarily affecting the human nervous system, underscoring the critical role of autophagy in brain function. We review the latest research delineating the landscape and network of autophagy in developing and mature neurons, elucidating how conserved autophagy pathways regulate neuronal homeostasis and functions at different ages. We examine the increasing evidence of dysfunctional autophagy that contributes to neurodevelopmental disorders, neurodegenerative diseases and psychiatric conditions. Recent insights into the dysregulation of autophagy provide valuable avenues for biomarker identification and therapeutic development, particularly targeting autophagy-lysosome pathways for neurological disorders.
    DOI:  https://doi.org/10.1038/s41593-026-02426-6
  7. Int J Mol Sci. 2026 Aug 21. pii: 7485. [Epub ahead of print]27(16):
      Mechanical stress is thought to be involved in the pathogenesis and pathophysiology of osteoarthritis (OA). However, much remains to be elucidated regarding how chondrocytes sense and respond to mechanical stress (stress sensing and response factors). Additionally, it still remains unclear whether there are defensive responses and mechanisms to protect against pathological agents and mechanical stress in articular cartilage tissue. This study was designed to determine whether repetitive mechanical force, at physiologic levels, affects the expression of factors regulating autophagy such as the autophagy-related proteins ATG5, Beclin-1, and Parkin, and the autophagy process as well as cellular activity in cultured chondrocytes. Three-dimensional cultured tissue was generated from human chondrocytes using a collagen sponge scaffold. After physiological mechanical loading of the 3D cell-collagen sponge construct, comparative analyses of expression levels of ATG5, Beclin-1, and Parkin were performed in human chondrocytes. Chondrocyte activity and Transmission Electron Microscopy (TEM) analysis for detecting autophagy process were also analyzed with or without repetitive compressive loading. In chondrocytes, 60 min or 180 min repetitive compressive loading significantly decreased the expression of ATG5, Beclin-1 and Parkin in comparison with the non-loading group. TEM analysis indicated that, in normal chondrocytes of the non-loading group, the autophagy process was shown to be progressing. In contrast, repetitive loading decreased the number of autophagosomes and autolysosomes in chondrocytes. In addition, numerous degenerated organelles that had not undergone autophagy were observed within the chondrocytes under repetitive loading. The ATG5 and Beclin-1 proteins are known to play crucial roles in regulating cellular autophagy. Furthermore, repetitive mechanical loading caused a decreased expression of Parkin, a mitophagy regulator in chondrocytes. Our results indicate for the first time that a decrease in mitophagy, as well as cellular autophagy, in response to mechanical stress, even at the physiologic level, leads to the accumulation of defective mitochondria and abnormal cellular proteins, resulting in reduced chondrocyte activity and affecting the maintenance of cartilage tissue homeostasis, ultimately contributing to the progression of OA.
    Keywords:  ATG5; Beclin-1; Parkin; autophagy; chondrocytes; mechanical stress; mitochondria; osteoarthritis
    DOI:  https://doi.org/10.3390/ijms27167485
  8. Circ Res. 2026 Aug 26.
       BACKGROUND: Protein quality control is critical for maintaining sarcomere structure and function in cardiomyocytes. Mutations in protein quality control pathway proteins, namely, CRYAB-R120G (arginine to glycine at position 120) and BAG3-P209L (proline to lysine at position 209), induce protein aggregates and cardiomyopathy in humans. Novel observations in yeast demonstrate mitochondrial uptake of cytosolic protein aggregates. We hypothesized that mitochondrial uptake of cytosolic protein aggregates, and their removal by mitophagy, a lysosomal degradative pathway, facilitates cytosolic protein quality control in cardiomyocytes.
    METHODS: Mice with inducible cardiac myocyte-specific ablation of TRAF2 (TNF receptor-associated factor 2; TRAF2-icKO), which impairs mitophagy, were assessed for protein aggregates with biochemical fractionation and super-resolution imaging. Human-induced pluripotent stem cell-derived cardiomyocytes with TRAF2 ablation or R120G knock-in to the CRYAB locus were assessed for protein aggregates and effects of mitophagy stimulation. Transgenic mice expressing R120G-CRYAB protein (R120G-transgenic mice) were subjected to adeno-associated virus 9-cTnT (cardiac troponin T) promoter-driven TRAF2 or PARKIN gain-of-function and TRAF2 loss of function in cardiomyocytes to determine the effect of mitophagy modulation on cardiac structure, function, and protein aggregate pathology.
    RESULTS: TRAF2-icKO mice demonstrate accumulation of mitochondrial and cytosolic protein aggregates and DESMIN mislocalization to protein aggregates. TRAF2 null human-induced pluripotent stem cell-derived cardiomyocytes demonstrate impaired mitophagy with accumulation of polyubiquitinated proteins and disrupted sarcomeres, which are rescued by both TRAF2 and PARKIN transduction. Isolated mitochondria take up cardiomyopathy-associated aggregate-prone cytosolic proteins, namely, R120G-CRYAB and P209L-BAG3. R120G-CRYAB mutant protein increasingly localizes to mitochondria in human and mouse cardiomyocytes. R120G-transgenic mice demonstrate upregulation of myocardial TRAF2 with increased mitophagy. Adult-onset inducible haplo-insufficiency of TRAF2 resulted in accelerated mortality, left ventricular systolic dysfunction, and increased protein aggregates in R120G-transgenic mice. Conversely, adeno-associated virus 9-TRAF2 transduction in R120G-transgenic mice stimulated mitophagy, reduced mortality, attenuated LV systolic dysfunction, reduced cytosolic protein aggregates, and restored DESMIN localization.
    CONCLUSIONS: Stimulation of mitophagy in cardiomyocytes facilitates removal of cytosolic protein aggregates as a mechanism to ameliorate proteotoxic cardiomyopathy.
    Keywords:  autophagosomes; heart failure; longevity; myocytes, cardiac; protein aggregates
    DOI:  https://doi.org/10.1161/CIRCRESAHA.126.328328
  9. J Biol Chem. 2026 Aug 28. pii: S0021-9258(26)02383-5. [Epub ahead of print] 113511
      Lysosomal storage disorders (LSDs) are intractable rare diseases caused by lysosomal dysfunction due mainly to defects in lysosomal enzyme genes. For many lysosomal enzymes to be transported correctly into lysosomes, their mannose-6-phosphate (M6P) labeling by GlcNAc-1 phosphotransferase (GNPT) is crucial. M6P-modified lysosomal enzymes are captured by M6P receptors and transported to lysosomes. The M6P-dependent pathway is utilized not only for newly synthesized lysosomal enzymes but also for the intracellular transport of exogenously administered lysosomal enzymes to lysosomes. In this study, we performed gene knockouts targeting the M6PR and IGF2R genes, which encode the M6P receptors. Cells with a double knockout of these genes secrete M6P-modified proteins into the culture supernatant (dKO sup), and we investigated its potential for therapeutic application. I-cell disease (mucolipidosis II, ML-II) arises from GNPT deficiency and is a disorder in which dozens of lysosomal enzymes are deficient within the cell. Treating ML II cells with the dKO sup restored lysosomal enzyme activity, reduced inclusion bodies, and improved autophagic function. ML II patient-derived fibroblasts exhibited increased lysosomal enzyme activities, normalized morphology, and improved cryo-viability with the treatment. NPC2 protein is not a lysosomal enzyme but functions cooperatively with NPC1 protein to mediate cholesterol transport in lysosomes. NPC2 deficiency causes Niemann-Pick disease type C2, and the dKO sup restored the cellular function of the disease by supplementing the NPC2 protein. These results indicate that M6PR/IGF2R double-deficient cells provide a simple platform for supplying M6P-modified proteins that can be applied to the treatment of a broad range of LSDs.
    Keywords:  GlcNAc-1-phosphotransferase; I-cell disease; Niemann-Pick C2; enzyme replacement therapy; lysosomal storage disorder; lysosome; mannose 6-phosphate; mucolipidosis II
    DOI:  https://doi.org/10.1016/j.jbc.2026.113511
  10. Cells. 2026 Aug 14. pii: 1459. [Epub ahead of print]15(16):
      HS-1-associated protein X-1 (HAX-1) is a ubiquitously expressed, multifunctional protein that regulates Ca2+ homeostasis and cell survival in cardiac muscle. In addition to its well-established anti-apoptotic function, HAX-1 has recently been implicated in autophagy regulation. In the present study, we explored the molecular mechanisms underlying HAX-1-mediated autophagy modulation in cellular models, including cardiac-derived H9c2 myotubes. HAX-1 overexpression enhanced autophagic activity, as evidenced by decreased sequestosome-1 (p62), increased microtubule-associated protein light chain 3-II (LC3-II), enhanced LC3 puncta formation, and elevated autophagic flux. Conversely, HAX-1 knockdown attenuated autophagic activity. Mechanistically, co-immunoprecipitation assays showed that HAX-1 associates with both p62 and LC3. Bioinformatic analysis of the HAX-1 protein sequence identified two conserved LC3-interacting region (LIR) motifs within its N-terminal domain. Deletion of this LIR-containing region (HAX-1ΔLIR) reduced LC3 association, decreased autophagic activity, and impaired autophagy-dependent clearance of HAX-1 itself following autophagy induction, suggesting the importance of these motifs. At a cardiac-relevant level, HAX-1 promoted a chloroquine-sensitive reduction in protein levels of its known binding partner, phospholamban (PLN), a key regulator of sarcoplasmic reticulum (SR) Ca2+ cycling. These findings indicate a previously unrecognized LC3/LIR-dependent mechanism underlying HAX-1-mediated autophagy modulation and suggest a potential role for HAX-1 in SR protein proteostasis.
    Keywords:  HAX-1; LC3; autophagy; cardiac cell; cardioprotection; p62; phospholamban
    DOI:  https://doi.org/10.3390/cells15161459
  11. FEBS Lett. 2026 Aug 27.
      Maintenance of protein homeostasis requires coordination between protein synthesis and degradation, yet whether autophagy directly controls translational machinery remains unclear. Here we explored whether autophagy maintains translational fidelity. Proteomic analysis of ubiquitin-enriched fractions and p62-associated proteins in autophagy-deficient tissue confirmed previous studies identifying ribosomal proteins and RNA-binding factors among candidate autophagy cargo. Pharmacological or genetic impairment of autophagy increased translational errors, particularly during oxidative stress. Components of the translational machinery localised to LC3-positive autophagic structures and accumulated in human brain tissue affected by neurodegeneration. These findings support a hypothesis in which autophagy preserves protein synthesis quality by removing damaged translational machinery. We term this proposed mechanism translophagy, potentially linking autophagy dysfunction, oxidative stress and the aberrant proteins in neurodegenerative disease.
    Keywords:  ageing; aggregation; autophagy; mRNA; neurodegeneration; oxidative stress; ribosome; translation
    DOI:  https://doi.org/10.1002/1873-3468.70448
  12. Front Cell Infect Microbiol. 2026 ;16 1835140
      Coxiella burnetii is a Gram-negative, obligate intracellular pathogen and the causative agent of the zoonotic disease Q fever. Resident alveolar macrophages are the first target cells, but C. burnetii spreads to other cell types. While we have information about C. burnetii uptake and the establishment of the replication-competent phagolysosomal-like C. burnetii-containing vacuole (CCV), it is not well studied how C. burnetii exits its host cell. Here, we show that an infection with C. burnetii also triggers the activation of TFEB, a master regulator of autophagy and lysosomal development. The activation occurs in a time-dependent manner and depends on the size of the CCV. Crucially, TFEB activation during C. burnetii infection depends on MCOLN1, which channels Ca2+ across the lysosomal membrane into the cytosol. Knock-down of MCOLN1 resulted in reduced TFEB activation and smaller CCVs, while MCOLN1 activation boosted bacterial egress. Indeed, peripheral CCVs are positive for LAMP1/2 and release bacteria, without inducing host cell death. Importantly, LAMP1/2 and C. burnetii were stainable in non-permeabilized cells at sites of bacterial release, demonstrating fusion of the lysosome with the plasma membrane. Crucially, while replication of C. burnetii is not inhibited in cells lacking LAMP1/2, egress is impaired. Taken together, our data indicates that with increasing CCV size, TFEB is activated by the release of Ca2+ from lysosomes via the MCOLN1 channel, which in turn enables further CCV development and damage of the CCV membrane. This triggers lysosomal exocytosis and egress of C. burnetii without cell death induction.
    Keywords:  Coxiella burnetii; Egress; LAMP; MCOLN1; Q fever; TFEB; lysosomal exocytosis; type IV secretion system
    DOI:  https://doi.org/10.3389/fcimb.2026.1835140
  13. Exp Eye Res. 2026 Aug 24. pii: S0014-4835(26)00371-4. [Epub ahead of print] 111215
      Age-related macular degeneration (AMD) is the most common blinding disease in the western world and is currently incurable. Although the exact causes of AMD are not clear, the primary origin of pathology appears to be the aged retinal pigment epithelium (RPE) exhibiting signs of lysosomal dysfunction and oxidative damage. RPE is responsible for the daily digestion of photoreceptor outer segments (POS), imposing a heavy continuous burden on the lysosomal network. A cellular model of RPE lysosomal dysfunction can be achieved by feeding RPE with a single pulse of POS, leading to the accumulation of autofluorescence granules (AFG), similar to lipofuscin in vivo. Here we show that synchronous phagocytosis of POS leads to early transient mTOR activation followed by inhibition in late phagosome maturation. One of its substrates, the transcription factor EB (TFEB) increases during phagosome maturation albeit mostly in its inactive phosphorylated form. We questioned whether modulation of the mTOR/TFEB axis could improve POS clearance and hence reduce AFG load. Treatment of POS-fed cells after the appearance of AFGs with rapamycin, an mTORC1 inhibitor results in ∼30% reduction of AFG load. This effect is dependent on active lysosomal enzymes and induction of active dephosphorylated TFEB with consequent activation of GADD34 and lysosomal biogenesis. As a proof of concept, we show that overexpressing a constitutively active form of unphosphorylated TFEB dramatically reduces POS-dependent AFG accumulation. Overall, this study suggests that viral or pharmacological approaches activating the TFEB pathway in the RPE could be beneficial as cell-protective treatment of early/intermediate cases of AMD, acting to delay progression of the disease.
    Keywords:  TFEB; autofluorescent granules; lysosomal dysfunction; mTOR; photoreceptor outer segments phagocytosis
    DOI:  https://doi.org/10.1016/j.exer.2026.111215
  14. Front Med (Lausanne). 2026 ;13 1905758
      Long COVID is frequently characterized by exertion intolerance, delayed symptom exacerbation, treatment sensitivity, and prolonged recovery. Post-exertional malaise (PEM) is often interpreted as a manifestation of low energy availability, autonomic dysfunction, immune activation, endothelial disturbance, or deconditioning. This paper proposes an additional recovery-failure mechanism for a PEM-dominant subgroup: fragile mitophagy, defined as a mismatch in which mitochondrial injury and mitophagy engagement occur but lysosomal completion of mitochondrial degradation is inadequate. Incomplete clearance could permit mitochondrial debris and danger signaling to persist and amplify oxidative, innate immune, endothelial, and neuroimmune responses after physiologic stress. The model predicts delayed crashes, progressive lowering of baseline with repeated exertion, and poor tolerance of interventions that increase mitochondrial turnover when lysosomal capacity is insufficient. Patient-derived Long COVID studies demonstrate mitochondrial, metabolic, and structural abnormalities but do not yet establish defective dynamic mitophagy flux or lysosomal completion. Hydroxychloroquine and chloroquine are used only as pharmacologic analogies showing that late autophagic flux can be impaired by disrupted lysosomal handling. Trehalose, genistein, curcumin, and the curcumin analog C1 are presented as experimental mechanistic probes because of reported effects on TFEB or autophagy-lysosome biology, not as established Long COVID treatments. The central prediction is that PEM severity will correlate more closely with impaired lysosomal completion, abnormal mitophagy flux, mitochondrial debris, and danger signaling than with baseline adenosine triphosphate (ATP) deficiency alone.
    Keywords:  endotype; fragile mitophagy; long COVID; lysosome; mitochondrial DNA; mitochondrial dysfunction; mitophagy; post-exertional malaise
    DOI:  https://doi.org/10.3389/fmed.2026.1905758
  15. Sci Adv. 2026 Aug 28. 12(35): eaec4519
      The physiological role of lipid asymmetry in intracellular membranes remains poorly understood. Here, we show that sphingomyelin (SM), typically confined to the lumen of the trans-Golgi network (TGN), is exposed on its cytoplasmic surface by the action of the Golgi-associated protein, Golgi-associated gamma-adaptin ear-containing adenosine 5'-diphosphate-ribosylation factor-binding protein 1 (GGA1). This exposure is driven by the GGA1 GAT domain, which induces lipid scrambling in a manner dependent on membrane curvature and cholesterol. SM exposure coincides with the exit of mannose 6-phosphate receptors from the TGN, a process essential for lysosomal enzyme trafficking. Furthermore, SM is transferred to autophagic membranes, where it facilitates autophagosome-lysosome fusion. These findings reveal a previously unrecognized role for lipid remodeling in membrane trafficking and autophagy.
    DOI:  https://doi.org/10.1126/sciadv.aec4519
  16. Nat Commun. 2026 Aug 28. pii: 9144. [Epub ahead of print]17(1):
      Membrane recruitment is a fundamental regulator of protein function. However, the allosteric mechanisms by which lipid binding controls protein activity remain poorly understood. In autophagy, the ubiquitin-like protein LC3 is lipid-anchored to autophagosomes, where it is essential for receptor recruitment and vesicle formation. While LC3-receptor interactions are structurally well defined, how membrane engagement governs LC3 functional dynamics has remained enigmatic. Here, we uncover that membrane binding triggers a major conformational transition in LC3, exposing functional pockets that are occluded in its cytosolic form. We demonstrate that this shift is mediated by dynamic coupling between the allosteric site (α3-loop5-β3-loop6) and the functional binding pockets. To conclusively test this mechanism, we utilised an ensemble-based protein design strategy guided by molecular dynamics to engineer the allosteric site. From a series of mutants, two variants emerged that stabilized LC3 conformation in either active or inactive state on the membrane. X-ray crystal structures of mutant LC3, biophysical assays, super-resolution microscopy, and TEM confirmed that the activated allosteric site mutant facilitates receptor binding and cargo capture. In contrast, the inactive variant is functionally inert on the membrane. Our work identifies a fundamental lipid-triggered allosteric site in LC3 that is critical for autophagy regulation and broader implications of membrane-dependent reprogrammable protein activities.
    DOI:  https://doi.org/10.1038/s41467-026-76697-9
  17. Chem Biomed Imaging. 2026 Aug 24. 4(8): 1740-1749
      Autophagic flux is a highly dynamic process essential for cellular homeostasis, yet its reliable visualization in live cells remains challenging due to the limitations of transfection-based LC3 reporters. Here, we present ATP3, a fluorogenic and ratiometric chemical probe that directly engages autophagic vesicles to enable the high-contrast and homogeneous imaging of autophagic flux without genetic manipulation. ATP3 constitutes a guanine targeting moiety and a smart fluorophore, with the former facilitating autophagic vesicle engagement, while the latter is intrinsically quenched and environment-responsive. ATP3 exhibits minimal background fluorescence and robust signal amplification upon autophagy-dependent engagement. Side-by-side comparisons demonstrate that ATP3 outperforms the conventional mRFP-GFP-LC3 assay in imaging contrast and staining uniformity. Furthermore, ATP3 enables dynamic monitoring of autophagic responses in an oxygen-glucose deprivation model, revealing progressive enhancement of autophagic flux under ischemic stress. Together, ATP3 provides a robust and broadly applicable chemical tool for visualizing autophagy dynamics in physiologically and disease-relevant contexts.
    Keywords:  autolysosome; autophagosome; autophagy; guanine; imaging; probe
    DOI:  https://doi.org/10.1021/cbmi.5c00285
  18. Autophagy Rep. 2026 ;5(1): 2721027
      As the site of photosynthesis and carbohydrate metabolism, chloroplasts are essential for phototrophic plant growth. The selective elimination of damaged chloroplasts by autophagy, a process known as chlorophagy, is crucial for chloroplast quality control. However, the mechanisms that activate chlorophagy and that underline its specificity remain poorly understood. Our recent study identified changes in maltose metabolism as a previously unrecognized trigger of a micro-chlorophagy program that depends on components of the core autophagy machinery. These findings provide a new starting point for uncovering the molecular principles that govern micro-chlorophagy. Here, we discuss emerging evidence showing that metabolic changes induce autophagic chloroplast turnover and outline the key challenges in defining the signals that initiate micro-chlorophagy.
    Keywords:  Arabidopsis; autophagy; chlorophagy; chloroplast; maltose metabolism; plant
    DOI:  https://doi.org/10.1080/27694127.2026.2721027
  19. MedComm (2020). 2026 Sep;7(9): e70911
      Apoptosis is a core program regulating organismal homeostasis and plays a pivotal role in the onset and progression of most diseases. Increasing evidence in recent years indicates that mitochondria are not only central to cellular metabolism but also play a pivotal role in regulating apoptosis. However, no systematic review elucidating how mitochondria finely regulate apoptotic processes through multidimensional mechanisms, including apoptosis-resistant diseases such as cancer. This paper systematically summarizes the molecular mechanisms by which mitochondria mediate apoptosis. We focus on the regulation of cytochrome c (Cyt c) release by the Bcl-2 protein family and the activation of downstream caspase cascades. Furthermore, we provide an in-depth analysis of intrinsic factors, including mitochondrial structural remodeling (membrane rupture, cristae remodeling, and membrane lipid redistribution), dynamics imbalance (fusion, fission, and mitophagy), and mitochondrial DNA abnormalities, as well as extrinsic factors involving interorganelle interactions with the endoplasmic reticulum, lysosomes, and other organelles. Additionally, we review clinical and preclinical advances in drugs targeting these pathways. This review aims to provide a comprehensive perspective on the complex network of mitochondrial regulation of apoptosis and offer valuable insights for developing novel clinical therapeutic strategies for cancer and other diseases.
    Keywords:  MAM; MOMP; apoptosis; mitochondria; mitochondrial autophagy; mitochondrial transplantation; mtDNA
    DOI:  https://doi.org/10.1002/mco2.70911
  20. Mol Med Rep. 2026 Oct;pii: 290. [Epub ahead of print]34(4):
      Hypertension‑induced cardiac fibrosis is a major risk factor for heart failure; although disrupted mitochondrial homeostasis has been confirmed to serve a critical role in the pathological process, its upstream regulatory factors remain incompletely understood. In the current study, RNA sequencing and bioinformatics analyses identified POU domain class 2 transcription factor 1 (POU2F1) as a hub transcriptional regulator in the fibrotic cardiac tissues of spontaneously hypertensive rats (SHRs). The expression levels of POU2F1 were associated with the severity of myocardial fibrosis, and cardiac expression of PTEN‑induced kinase 1 (PINK1) and Parkin in SHRs. Complementing these in vivo observations, angiotensin II stimulation significantly upregulated POU2F1 expression in cardiac fibroblasts (CFs) in vitro. Furthermore, POU2F1 expression exhibited a positive correlation with fibroblast activation, as indicated by α‑smooth muscle actin fluorescence intensity. Mechanistically, POU2F1 knockdown attenuated CF activation, improved mitochondrial structure and energy metabolism, and restored PINK1/Parkin‑mediated mitophagy balance in vivo and in vitro. Conversely, POU2F1 overexpression was associated with enhanced PINK1/Parkin‑mediated mitophagy signaling. Crucially, through chromatin immunoprecipitation‑quantitative PCR, electrophoretic mobility shift assay and dual‑luciferase reporter assay, it was demonstrated that POU2F1 can directly bind to the PINK1 promoter to activate its transcription. In conclusion, the present study identified a novel role for POU2F1 in hypertensive cardiac fibrosis, demonstrating that it exacerbates disease progression by disrupting mitochondrial homeostasis through transcriptional activation of PINK1, accompanied by alterations consistent with enhanced PINK1/Parkin‑mediated mitophagy.
    Keywords:  PINK1; POU2F1; cardiac fibrosis; hypertension; mitophagy
    DOI:  https://doi.org/10.3892/mmr.2026.14001
  21. Cell Death Differ. 2026 Aug 26.
      The RNA binding G3BP1 is depleted in several neurodegenerative diseases, yet its functional consequences at the cellular level remain poorly understood. While best known for its critical role in stress granule formation, we demonstrate that G3BP1 also stabilises the COPI vesicle protein beta-COP by promoting its interaction with the deubiquitinase USP10. G3BP1 depletion disrupts this interaction leading to increased ubiquitination of beta-COP, which accelerates its proteasomal degradation. This leads to compromised Golgi structure and function, and impaired lysosomal homeostasis, which causes defective autophagic flux. Consequently, the autophagic clearance of α-synuclein, a protein that can drive Parkinson's disease (PD), is significantly slowed. Importantly, we observe a concurrent reduction of both G3BP1 and beta-COP protein levels in brain sections from PD and dementia with Lewy Body (DLB) patients and from a PD mouse model. These findings reveal a novel mechanistic link between G3BP1, vesicular trafficking, and proteostasis in neurodegeneration.
    DOI:  https://doi.org/10.1038/s41418-026-01853-z
  22. Nat Commun. 2026 Jul 24. pii: 9041. [Epub ahead of print]17(1):
      Autoimmune thyroiditis arises from disrupted homeostasis of thyroid follicular epithelial cells and coordinated immune cell activation within the microenvironment. However, its pathogenesis is not fully understood. Here, we identify a mitochondrial (mt) DNA-cGAS-STING inflammatory axis as a driver of autoimmune thyroiditis in mice. By contrast, ubiquitin-dependent mitophagy mediated by PINK1 and Parkin was found to protect mice from disease. Mechanistically, mitochondrial dysfunction elevates mitochondrial reactive oxygen species levels, activating the ATM-CHK2 DNA damage response pathway, which in turn phosphorylates the autophagy adapter TAX1BP1 at Ser722. This modification promotes the recruitment of mitochondria to autophagosomes, thereby facilitating mitophagy. Impairing the ATM-CHK2-TAX1BP1 mitophagy pathway causes mtDNA leakage into the cytosol and triggers cGAS-STING-dependent inflammation. Notably, pharmacological inhibition of STING with C176 effectively slows autoimmune thyroiditis progression. Together, these findings define an mtDNA-driven pathogenic mechanism in autoimmune thyroiditis and identify STING as a potential therapeutic target.
    DOI:  https://doi.org/10.1038/s41467-026-76047-9
  23. Sci Adv. 2026 Aug 28. 12(35): eaee4940
      Neurons actively shape immune responses that maintain central nervous system integrity. We identify SPP1 (secreted phosphoprotein 1) as a neuron-derived signal that reprograms microglia into a neuroprotective, homeostatic state after injury and during neurodegeneration. In mouse models of glaucoma and optic nerve damage, neuronal SPP1 enhances microglial autophagy, debris clearance, and anti-inflammatory activity, preserving neuronal survival and visual function. SPP1 is elevated in neurons of human and primate glaucomatous retinas, where SPP1+ cells show increased resilience. In Alzheimer's disease brain, neuronal SPP1 correlates with neuronal survival, while microglia around Aβ plaques display defective autophagy. In human iPSC co-cultures, SPP1 enhances microglial Aβ clearance and prevents neurodegeneration. Thus, SPP1 defines a protective neuron-microglia axis in glaucoma and possibly other neurodegenerative diseases.
    DOI:  https://doi.org/10.1126/sciadv.aee4940
  24. J Cardiovasc Dev Dis. 2026 Aug 13. pii: 387. [Epub ahead of print]13(8):
      Pulmonary arterial hypertension (PAH) is a severe cardiovascular disease characterized by progressively increased pulmonary vascular resistance and right heart failure. Its pathogenesis involves multiple factors, including genetic predisposition, inflammation, oxidative stress, and imbalances between cell proliferation and apoptosis. Recent studies indicate that autophagy has a context-dependent dual role in PAH. Flux-competent autophagy may be protective by clearing damaged mitochondria, limiting excessive inflammation, and maintaining metabolic homeostasis, whereas excessive autophagy initiation or impaired autophagosome-lysosome degradation may promote metabolic dysfunction, inflammatory signaling, abnormal vascular cell phenotypes, and pulmonary vascular remodeling. This focused narrative review summarizes the molecular mechanisms and key signaling pathways linking autophagy to PAH, with emphasis on PTEN-induced kinase 1 (PINK1)/Parkin-mediated mitophagy and the AMP-activated protein kinase (AMPK)/mechanistic target of rapamycin (mTOR) energy-sensing axis. It also evaluates potential therapeutic strategies targeting key nodes of autophagy, such as AMPK activators and mTOR inhibitors, along with their clinical research progress. Finally, this review provides an outlook on future research directions, emphasizing the need to further elucidate the dynamic regulatory mechanisms and cell-type specificity of autophagy in order to advance the clinical translation of autophagy-targeted precision therapies for PAH.
    Keywords:  AMPK/mTOR; PAH; autophagy; mitophagy; pulmonary arterial hypertension; therapeutic targets; vascular remodeling
    DOI:  https://doi.org/10.3390/jcdd13080387
  25. Nat Commun. 2026 Jul 28. pii: 9165. [Epub ahead of print]17(1):
      Parkinson's disease (PD) is defined pathologically by loss of dopamine-producing neurons in the substantia nigra pars compacta (SNc). Yet synaptic dysfunction emerges much earlier, making it essential to define the mechanisms that drive early nigrostriatal deregulation. In the SNc, molecularly distinct dopamine neuron subtypes show differential susceptibility to PD. Here, we used intersectional genetic mouse models to determine how the PD-linked kinase LRRK2 affects vulnerable dopamine subtypes. Immunofluorescence and proximity-labeling proteomics revealed enriched LRRK2 expression in vulnerable dopamine neuron subclusters. High-resolution imaging showed that pathogenic LRRK2 disrupts presynaptic release-site organization in vulnerable dopamine axons, leading to reduced spontaneous and evoked striatal dopamine release in vivo. Proteomic analyses further showed that mutant LRRK2 increases phosphorylation of RAB3 proteins, impairing their interaction with the active-zone effectors RIM1 and RIM2. Together, these findings highlight a subtype-specific, cell-autonomous mechanism by which pathogenic LRRK2 impairs PD-vulnerable nigrostriatal synapses and provide a framework for therapeutic strategies targeting early synaptic deficits in PD.
    DOI:  https://doi.org/10.1038/s41467-026-75194-3
  26. Autophagy. 2026 Aug 23. 1-3
      In dividing cells, centrosomes are the primary microtubule-organizing centers and typically duplicate only once per cell division cycle, ensuring that there is just a single centrosome at each mitotic spindle pole. Centrosome amplification presents a paradox: although supernumerary centrosomes can induce chromosomal instability and promote invasive behavior, they may also impede cell proliferation by activating the Hippo-LATS pathway and the PIDDosome-p53 axis. Therefore, cells with supernumerary centrosomes must either tolerate the abnormal centrosome numbers or restore centrosome homeostasis. To identify pathways that allow cell proliferation in the presence of extra centrosomes, we conducted a genome-wide CRISPR/Cas9 screen in mouse embryonic stem cells to identify gene knock-outs that restored cell proliferation following induction of a polo-like kinase 4 (PLK4) transgene. In addition to components of known pathways that respond to supernumerary centrosomes, the screen identified a previously unrecognized Rho GTPase signaling network. Depletion of the RAC1 GTPase-activating protein ARHGAP15 increases autophagic flux, reduces the percentage of cells with supernumerary centrosomes in an ATG16L1-dependent manner, and restores proliferation of cells following PLK4 overexpression. Our findings support a model in which the centrosomal protein CEP170 promotes ARHGEF2-dependent RAC1 activation, enabling RAC1-GTP to interact with ATG16L1 and stimulate autophagy. ARHGAP15 counteracts this pathway, thereby limiting RAC1-ATG16L1 signaling and autophagy. Thus, our findings identify a mechanism that, in contrast to the Hippo-LATS and PIDDosome-p53 pathways, does not circumvent the effects of centrosome amplification by disabling cell-cycle arrest pathways but activates a corrective mechanism to reduce centrosome number.
    Keywords:  ARHGAP15; ATG16L1; RAC1; autophagy; centrosome amplification
    DOI:  https://doi.org/10.1080/15548627.2026.2719427
  27. Autophagy Rep. 2026 ;5(1): 2715823
      Pancreatic ductal adenocarcinoma (PDAC) cells rely on autophagy to adapt to microenvironmental stress. In our recent work, we showed that baseline autophagy flux levels shape the proliferative capacity of human PDAC cells. Here, we extend these findings by demonstrating that low baseline autophagy is consistently associated with enhanced proliferation in vitro and in vivo across multiple human and murine PDAC models. While this inverse relationship between autophagy flux and proliferation is conserved across species, the underlying regulatory mechanisms diverge. Namely, changes in baseline autophagy levels relied predominantly on canonical nutrient-sensing pathways in murine but not human PDAC cells. Additionally, we found that autophagy flux was regulated independently of the cell cycle or the genetic status of p53 (i.e., allelic deletion, point mutation). Collectively, our results reveal conserved phenotypic outcomes but divergent mechanisms, underscoring the need for parallel comparative approaches before extrapolating findings from mouse PDAC models to human disease.
    Keywords:  Autophagy; cancer; cell signaling; cross-species; proliferation
    DOI:  https://doi.org/10.1080/27694127.2026.2715823
  28. J Cardiol. 2026 Aug 28. pii: S0914-5087(26)00174-7. [Epub ahead of print]
      Dilated cardiomyopathy (DCM) is characterized by progressive ventricular dilation and systolic dysfunction, yet the cellular damage that precedes intercellular fibrosis remains incompletely defined. Electron microscopy has long revealed autophagic vacuoles and myofilament lysis in cardiomyopathy, but their pathological significance has been debated for decades. Recent advances in ultrastructural analysis, including LC3 immunoelectron microscopy, have enabled more precise characterization of these changes and have renewed interest in the role of autophagy in cardiomyopathy. In this review, we integrate ultrastructural findings from a large cohort of patients with DCM with contemporary clinical imaging data to clarify the sequence of myocardial damage. Myofilament lysis consistently appears as the earliest structural abnormality, preceding intercellular fibrosis and ventricular remodeling. Autophagic vacuoles are frequently observed at sites of lysis, suggesting activation of a conserved cellular repair response rather than a mechanism of cell death. This interpretation is supported by external evidence, including LC3-based analyses demonstrating associations between autophagy and reverse remodeling, as well as experimental studies showing that sodium-glucose cotransporter 2 inhibitors enhance stress-adaptive pathways and mitochondrial energetics even in models with impaired mitophagy. By integrating electron microscopy with late gadolinium enhancement (LGE) on cardiac magnetic resonance imaging, we further demonstrate that autophagic activity provides prognostic information beyond conventional tissue characterization. In LGE-negative myocardium, the presence of autophagic vacuoles identifies a subgroup with a lower risk of heart-failure recurrence, whereas myofilament lysis alone shows limited prognostic value. These findings highlight the importance of distinguishing structural damage from cellular repair activity when evaluating cardiomyopathy. Collectively, the evidence supports a refined framework in which myofilament lysis precedes intercellular fibrosis, and autophagy represents an adaptive response that modulates clinical outcomes. Understanding these ultrastructural processes may inform future diagnostic strategies and therapeutic approaches for cardiomyopathy.
    Keywords:  Autophagy; Cardiomyocyte; Dilated cardiomyopathy; Myocardial degeneration; electron microscopy
    DOI:  https://doi.org/10.1016/j.jjcc.2026.08.006
  29. Angew Chem Int Ed Engl. 2026 Aug 26. e7042721
      Macroautophagy is central to cellular homeostasis and emerges as a promising avenue for targeted degradation. However, there is still a lack of efficient approaches allowing for visualizing macroautophagic flux and degradation. Here we develop a semi-synthetic LC3-interacting degrader (SLID) that enables fluorogenic imaging of macroautophagic activities and visualization of targeted degradation. SLID is engineered by fusing LC3-interacting regions (LIRs) to a self-labeling tag and an oligomeric motif, with the LIRs for binding to autophagosomes, the oligomeric motif for enhancing the binding, and the self-labeling tag for visualizing autophagosome formation using a pH indicator. SLID is further coupled to an antibody domain through a dimerization-induced proximity system, allowing chemically inducible degradation of target proteins. We show that this SLID platform permits efficient degradation of diverse target proteins such as disease-associated aggregation-prone proteins and phase-separated condensates. SLID also reveals elevated macroautophagic activities in senescent cells, and is repurposed for inducing senescent cell apoptosis via degradation of pro-survival and anti-apoptotic proteins. Our study highlights the great promise of SLID as a versatile tool for studying macroautophagy and targeted protein degradation.
    Keywords:  apoptosis; biosensors; macroautophagy imaging; macroautophagy‐targeted degradation; senescent cells
    DOI:  https://doi.org/10.1002/anie.7042721
  30. eNeuro. 2026 Aug 25. pii: ENEURO.0116-26.2026. [Epub ahead of print]
      Peripheral neuropathy affects over 18 million adults in the U.S. (Hicks et al 2021), but therapeutic outcomes are poor due to a lack of regenerative treatments. Development of novel, disease-modifying therapies is hindered by poor understanding of the mechanisms promoting axonal regeneration in peripheral neurons. Pten is a strong negative regulator of cell growth, and Pten-KO drives axonal regeneration in various neuronal subtypes potentially via downstream regulation of stability of the microtubule (MT) cytoskeleton, a vital component of axonal growth. While Pten-KO accelerates MT polymerization rates in the axonal growth cone, it remains unknown whether this action is dependent on mTORC1 or mTORC2 signaling, and whether regeneration under Pten-KO is dependent on MT activation. Here, we perform in vitro codeletions of either Raptor (mTORC1) or Rictor (mTORC2) alongside Pten-KO in mouse peripheral sensory neuron cultures of either sex to isolate the effects of each pathway on the MT cytoskeleton. We use Pten-KO to increase MT polymerization and neuronal outgrowth, and then show that suppression of mTORC2, but not mTORC1, is sufficient to reduce the accelerated MT polymerization and neuronal hypertrophy to wild-type levels. These results are specific to the axonal growth cone, and MT dynamics in the proximal axon shaft are not impacted by Pten-KO, mTORC1 suppression, or mTORC2 suppression. Our results help elucidate the mechanism by which Pten regulates the MT cytoskeleton and axonal outgrowth in peripheral sensory neurons, localize where this occurs in the axon, and highlight the MT cytoskeleton as a potential molecular target for regenerative therapies.Significance statement Pten-KO promotes neuron growth, but Pten has several downstream effectors, and it is unclear which effectors drive axonal regeneration. One of these downstream targets is the microtubule cytoskeleton, which is vital for normal axonal growth and development. We demonstrate that Pten-KO increases MT polymerization rates in the growth cone, but not proximal shaft, in axons of peripheral sensory neurons. This effect is reversed by co-deletion of mTORC2, but not mTORC1, and the loss of mTORC2 primarily inhibits elongation of the distal axon, suggesting that the regulation of MT polymerization is specific to the growth cone. These results highlight MT dynamics in the axonal growth cone as a vital component of peripheral neuron regeneration and a potential therapeutic target.
    DOI:  https://doi.org/10.1523/ENEURO.0116-26.2026
  31. Front Cardiovasc Med. 2026 ;13 1929329
      Primary cilia are microtubule-based sensory organelles located on the cell surface and function as cellular signaling antennae for mechanical and chemical cues. In the vascular endothelium, they participate in shear-stress sensing, Ca2+ signal transduction, eNOS activity regulation and vascular homeostasis. Autophagy is a conserved lysosome-dependent degradative pathway that maintains cardiovascular cell homeostasis by removing damaged organelles, regulating lipid metabolism, restraining inflammatory activation and preserving proteostasis. In this review, we discuss the bidirectional regulatory relationship between primary cilia and autophagy. Primary cilia may influence autophagic activity through Ca2+/AMPK/mTOR, Hedgehog and PI3KC2α-related pathways, whereas autophagy can reciprocally regulate ciliary homeostasis through cilia-related proteins such as OFD1 and IFT20. This cilium-autophagy interface may contribute to hypertension, atherosclerosis and aortic aneurysm by modulating endothelial mechanosensing, mitochondrial quality control, lipid handling, ROS homeostasis and inflammatory responses. Because direct evidence for causal cilium-autophagy crosstalk in the cardiovascular system remains limited, we distinguish established mechanisms from biologically plausible extrapolations and disease-associated observations. By integrating evidence from ciliary mechanosensing, autophagy regulation and vascular pathological remodeling, this review proposes the primary cilium-autophagy axis as a conceptual framework linking hemodynamic disturbance, organelle stress and vascular disease progression, and provides a basis for refining mechanistic hypotheses and potential therapeutic targets.
    Keywords:  aortic aneurysm; atherosclerosis; autophagy; cardiovascular disease; hypertension; primary cilium; vascular disease
    DOI:  https://doi.org/10.3389/fcvm.2026.1929329
  32. Acta Neuropathol Commun. 2026 Aug 19. pii: 178. [Epub ahead of print]14(1):
      Microglial hyperactivation contributes to Parkinson's disease (PD) progression, yet the upstream microenvironmental cues that sustain this state remain incompletely understood. While α-synuclein (α-Syn) aggregation is a primary trigger, aging and PD are also associated with microvascular and perfusion abnormalities. However, how vascular-associated hypoxic stress interacts with protein toxicity in microglial fate determination remains unclear. We integrated human single-nucleus RNA sequencing (snRNA-seq) data, a chronic progressive transgenic mouse model (9-month-old A53T), and an in vitro "double-hit" model. Neuropathological and immunofluorescence analyses were employed to assess the neurovascular unit and microglial phenotypes. The snRNA-seq analysis of human PD brains revealed a Disease-Associated Microglia (DAM) subset characterized by enrichment of hypoxia and glycolysis pathways, with HIF1A acting as a central node. In vivo, 9-month-old A53T mice exhibited motor deficits and dopaminergic degeneration, accompanied by reduced CD31+ microvascular coverage in the substantia nigra. This reduction in CD31+ vascular coverage was associated with microglial HIF1A accumulation and increased IBA1-defined soma area. In vitro, physical hypoxia amplified α-Syn preformed fibril (PFF)-induced microglial reactivity, intracellular accumulation of phosphorylated α-Syn (p-αSyn). Our study supports a "double-hit" model in which hypoxia-associated stress may amplify α-Syn-induced microglial dysfunction through HIF1A-linked metabolic remodeling and impaired autophagy-related protein handling. Targeting neurovascular-immune interactions may offer therapeutic opportunities for advanced PD.
    Keywords:  Autophagic dysfunction; Disease-associated microglia; Hypoxia; Neurovascular unit; Parkinson's disease; single-nucleus RNA sequencing
    DOI:  https://doi.org/10.1186/s40478-026-02412-w
  33. Elife. 2026 Aug 27. pii: RP106865. [Epub ahead of print]14
      Endolysosomal dysfunction is a hallmark of Alzheimer's disease and related tauopathies, yet underlying mechanisms remain poorly understood. This study investigates the role of sphingolipid metabolism in maintaining endolysosomal membrane integrity and its impact on tau aggregation and toxicity in Caenorhabditis elegans and human cell culture models. Fluorescence recovery after photobleaching and C-Laurdan dye imaging revealed that silencing sphingolipid metabolism genes reduced endolysosomal vesicle membrane fluidity, increasing their rupture. The accumulation of aggregated tau in endolysosomal vesicles further aggravated endomembrane rigidification and damage, and promoted seeded tau aggregation, potentially by facilitating the escape of tau seeds from the endolysosomal system. Supplementation with unsaturated fatty acids improved membrane fluidity, suppressing endolysosomal rupture and seeded tau aggregation in cell models, and alleviating tau-associated neurotoxicity in C. elegans. Together, this study provides mechanistic insight into how perturbation of sphingolipid metabolism promotes endolysosomal membrane damage and contributes to the escape of aggregated tau from this compartment, suggesting that restoration of membrane fluidity may represent a strategy to limit tau propagation and toxicity.
    Keywords:  Alzheimer's disease; C. elegans; cell biology; endolysosomal system; lipid homeostasis; membrane fluidity; sphingolipid metabolism; tau propagation
    DOI:  https://doi.org/10.7554/eLife.106865