bims-axbals Biomed News
on Axonal biology and ALS
Issue of 2026–05–24
thirty-one papers selected by
TJ Krzystek



  1. Cell Death Dis. 2026 May 17.
      Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) are devastating neurodegenerative diseases linked by similar pathological mechanisms, which, in some familial forms, may be associated with the same genetic alterations. Among them, the most common is the C9ORF72 (C9) mutation. The C9 mutation consists in an aberrant expansion of the hexanucleotide repeat (G4C2)n that leads to the production and accumulation of toxic dipeptide repeat proteins (DPRs). Some of these C9-DPRs contribute to neuronal dysfunction and degeneration through different mechanisms. One of these involves alterations in the protein quality control (PQC) system, specifically in the autophagy-lysosomal pathway. Valosin-containing protein (VCP) is a critical component of the PQC system, assisting the degradation of misfolded proteins and damaged organelles and the maintenance of cellular homeostasis. In this study, we investigated the role of VCP in modulating pathological features associated with C9 mutation. Using neuronal cell models, we demonstrated that VCP overexpression significantly reduced C9-DPRs levels. This reduction is mediated by mechanisms involving both the ubiquitin-proteasome system (UPS) and autophagy. Additionally, we also observed that C9-DPRs induce lysosomal damage, which is counteracted by VCP overexpression, as indicated by decreased galectin-3 puncta and restored lysosomal pH. We then pharmacologically activated VCP-mediated clearance through SMER28, increasing the clearance of the most toxic DPR, the polyPR. We also determined that in this model, SMER28 activity is mediated by the UPS and is associated with the mitigation of DPR-induced lysosome damage. Additionally, using motor neurons derived from induced pluripotent stem cells (iPSC-MNs) from C9-ALS mutation carriers, we demonstrated that SMER28 treatment significantly decreased polyGA levels, a marker for C9-DPR accumulation. Moreover, SMER28 rescued C9-MNs commitment to differentiation and the alteration in the expression of autophagy-related genes. Taken together, our findings strongly support VCP as a modulator of C9 pathology and highlight its potential as a therapeutic target.
    DOI:  https://doi.org/10.1038/s41419-026-08856-1
  2. Cell Rep. 2026 May 21. pii: S2211-1247(26)00442-0. [Epub ahead of print]45(6): 117364
      Parkinson disease (PD)-associated mutations in the LRRK2 gene hyperactivate LRRK2 kinase activity, leading to increased phosphorylation of a subset of RAB GTPases, which are master regulators of intracellular trafficking. In neurons, processive retrograde transport of autophagosomes is essential for autophagosome maturation and effective degradation of autophagosomal cargo in the axon. Here, we show that knockout of the LRRK2-counteracting RAB phosphatase PPM1H causes a gene-dose-dependent disruption of the axonal transport of autophagosomes, leading to impaired degradation of axonal alpha-synuclein (aSyn), a key protein in PD pathophysiology. Defective autophagosome transport and impaired aSyn degradation correlate with increased aSyn aggregation in primary PPM1H knockout neurons exposed to preformed fibrils of aSyn, an effect that is dependent on LRRK2 kinase activity. These findings mechanistically link LRRK2-mediated RAB hyperphosphorylation to defective autophagosomal degradation and enhanced aggregation of aSyn, positioning the LRRK2-RAB axis as a key driver of PD pathophysiology.
    Keywords:  CP: Cell biology; CP: Neuroscience; LRRK2; PPM1H; RAB GTPases; alpha-synuclein; autophagy; axonal transport
    DOI:  https://doi.org/10.1016/j.celrep.2026.117364
  3. bioRxiv. 2026 May 05. pii: 2026.04.30.722019. [Epub ahead of print]
      Mitochondrial (MT) dysfunction is a key driver of ALS pathology. Without a healthy MT system, motor neurons (MN) function at sub-optimal levels and die. In addition, other effects of ALS, like axon/dendrite degeneration, may occur from a pathophysiological cascade spurred by MT dysfunction. A phenotypic screen identified Dipyridamole (DPM), an FDA-approved and safe drug, as having extraordinary effects on ALS patient induced pluripotent stem cell (iPSC)-derived MNs. The drug prevented MT fragmentation, loss of MT content, impaired MT bioenergetics, axon/dendrite degeneration, and premature MN death, extending neuronal survival by more than fivefold. Importantly, its efficacy extended across iPSC-derived neurons representing two different familial forms of ALS (C9orf72, TDP43) and Alzheimer's disease (PSEN1), implying broad neuroprotection across ALS forms and other neurodegenerative diseases. DPM increased MT respiration and pyruvate uptake in a mechanism requiring the Mitochondrial Pyruvate Carrier (MPC), mechanistically explaining its biological activities. Thus, DPM is a promising drug to repurpose or refine for treating neurodegenerative diseases or other diseases that would benefit by augmenting pyruvate uptake into MT.
    Teaser: Dipyridamole, an FDA-approved drug, restores mitochondrial function and protects neurons in ALS and Alzheimer's disease.
    DOI:  https://doi.org/10.64898/2026.04.30.722019
  4. Jpn J Radiol. 2026 May 18.
      Neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), Parkinson's disease (PD) and Huntington's disease (HD) cause progressive loss of specific neuronal populations and currently lack curative therapies. Animal models and immortalized cell lines incompletely recapitulate human pathology and genetic heterogeneity, limiting drug discovery. Human induced pluripotent stem cells (iPSCs) provide a patient‑specific platform for disease modelling, drug screening and studying individual responses. Translational research (TR) uses iPSC models to identify candidate therapies that are subsequently tested in clinical trials, while reverse translational research (rTR) feeds clinical observations back to the bench by analyzing iPSCs derived from trial participants and integrating molecular data with patient phenotypes. This review summarizes recent advances in iPSC‑based TR and rTR for ALS and extends the discussion to other neurodegenerative diseases. Key clinical trials launched from iPSC screens-ropinirole, retigabine and bosutinib-are reviewed alongside emerging rTR efforts that use patient‑derived iPSCs to identify biomarkers and therapeutic mechanisms. We also survey iPSC models for AD, PD and HD, highlighting applications of three‑dimensional (3D) brain organoids and gene‑editing technologies. Finally, we discuss future directions for precision medicine, multimodal integration and technological challenges, with particular attention to how imaging biomarkers may complement iPSC-based TR/rTR frameworks in neurodegenerative diseases.
    Keywords:  Amyotrophic lateral sclerosis (ALS); Imaging biomarkers; Induced pluripotent stem cells (iPSCs); Precision medicine; Translational and reverse translational research; iPSC-based drug discovery
    DOI:  https://doi.org/10.1007/s11604-026-02000-x
  5. BMC Biol. 2026 May 19.
       BACKGROUND: Axon initial segment (AIS) dysfunction disrupts neuronal compartmentalization, which leads to pathological processes like Tau missorting in Alzheimer's disease (AD). However, the molecular mechanisms that destabilize the AIS scaffold are incompletely understood. Our group has previously shown that the Abl1 non-receptor tyrosine kinase is aberrantly activated in AD mouse models and promotes dendritic spine collapse, Tau hyperphosphorylation, and neuronal apoptosis. Given the important role of Abl1 in AD and its emerging significance in Tau pathology, we examined how it contributes to AIS collapse.
    RESULTS: We find that activation of Abl1 by amyloid-β fibrils promotes AIS disruption, as determined by the loss of clustered AnkG in the proximal axon, and that this can be prevented by pharmacological inhibition of Abl kinases. Cytosolic extraction experiments show that active Abl1 associates to the AIS scaffold, and this association increases in response to amyloid-β fibril treatment. Furthermore, using expansion microscopy, we show that Abl1 localizes to the AIS in dissociated hippocampal cultures and in mouse brain slices. We find a decrease in AIS actin patches, key for maintenance of neuronal compartmentalization, following Abl kinase activation. Finally, we show that Abl1 activation promotes missorting of somatodendritic Rab11 into the axon and the axonal protein Tau into the somatodendritic compartment, indicating a bidirectional failure in AIS barrier function.
    CONCLUSIONS: Taken together, our results show that Abl1 plays an important role in AIS destabilization and that its activation compromises protein compartmentalization in a primary neuron culture model of AD.
    Keywords:  Abl kinases; Actin; Alzheimer’s disease; Axon initial segment; Protein sorting
    DOI:  https://doi.org/10.1186/s12915-026-02624-5
  6. bioRxiv. 2026 May 05. pii: 2026.04.30.722110. [Epub ahead of print]
      Gain-of-function mutations in LRRK2 are a major cause of inherited Parkinson's disease. LRRK2 encodes a multidomain kinase, whose bidirectional interplay with Rab GTPases regulates critical cellular processes like lysosomal homeostasis. Certain Rabs, including Rab12 and Rab29, recruit LRRK2 to organelle membranes and stimulate its kinase activity; activated LRRK2 phosphorylates a subset of Rabs in their Switch-II motifs. Molecular basis governing selective Rab recognition by LRRK2 remains unclear. Here we structurally characterize LRRK2 interactions with representative Rab GTPases and identify three novel Rab-binding sites: site 4 for Rab8A/10, site 5 for Rab43, and site 6 for Rab5A, defining a total of six distinct binding sites that account for known LRRK2-interacting Rabs. Additionally, we elucidated the binding site of GABARAP, an ATG8 member that recruits LRRK2 to stressed lysosomes. Our findings provide a framework for therapeutic targeting of LRRK2 recruitment for Parkinson's.
    DOI:  https://doi.org/10.64898/2026.04.30.722110
  7. bioRxiv. 2026 May 09. pii: 2026.05.06.723191. [Epub ahead of print]
      Septins are cytoskeletal filaments that associate with the actin and microtubule cytoskeleton, but the mechanisms that govern septin crosstalk and function with these networks are largely unknown. Here, we show that glycogen synthase kinase 3 (GSK3) directly phosphorylates septin-9 (SEPT9), acting as a molecular switch that bidirectionally controls septin distribution between actin and microtubules. We show that GSK3 inhibition redistributes endogenous SEPT9 toward microtubules in multiple cell types. Phosphomimetic mutations at serines 82 and 85 reduce microtubule binding and enhance actin association in cells and in vitro, while phosphonull mutations promote microtubule binding and growth. In primary hippocampal neurons, GSK3β inactivation promotes SEPT9-microtubule association, and phosphomimetic mutations impair asymmetric neurite growth during neuronal polarization. These findings reveal a phosphorylation-dependent mechanism of septin partitioning between actin and microtubules, placing the cytoskeletal functions of septins under the control of GSK3 - a kinase linked to multiple signaling pathways of cell physiology and metabolism.
    Highlights: GSK3β phosphorylates SEPT9, and its activity gates septin-cytoskeleton associationS82/S85 phosphorylation reduce microtubule binding and increase actin localizationUnphosphorylated SEPT9 binds preferentially to microtubules, promoting their growthGSK3β inactivation drives SEPT9 to microtubules to establish neuronal polarity.
    DOI:  https://doi.org/10.64898/2026.05.06.723191
  8. Cell Rep Methods. 2026 May 19. pii: S2667-2375(26)00157-8. [Epub ahead of print] 101457
      Degeneration of the corticospinal tract is a feature in several neurodegenerative disorders and leads to disability. However, modeling corticospinal neuron (CSN) pathology and corticospinal connectivity is challenging, as there are interspecies differences in these networks. We developed a human induced pluripotent stem cell (hiPSC)-based microfluidic platform for modeling human CSN and spinal motor neuron (SpMN) connectivity. The incorporation of regionally specific astrocyte subtypes (cortical and spinal) in addition to CSNs and SpMNs allows for the modeling of neural cell interactions. Multielectrode array electrophysiology reveals the temporal maturation of the network. Retrograde labeling demonstrates synaptic connectivity between CSNs and SpMN. Optogenetic strategies to selectively activate excitatory cortical neurons (CNs) attenuated by glutamate receptor antagonism confirm the functional relevance of the model. Incorporating morphological, electrophysiological, and physiological measures of corticospinal connectivity, this platform is a versatile model for neurodegenerative disease research and the future development of targeted CSN therapies.
    Keywords:  CP: neuroscience; CP: stem cell; cortical neuron; corticofugal; motor neuron; stem cells
    DOI:  https://doi.org/10.1016/j.crmeth.2026.101457
  9. Commun Biol. 2026 May 19.
      Perineuronal nets (PNNs) are extracellular matrix structures that stabilise synaptic inputs and play a role in regulating neuronal plasticity. Although PNN dysregulation is observed in several neurological disorders, their relevance to amyotrophic lateral sclerosis (ALS) remains unclear. In particular, the extent to which PNN alterations reported in ALS animal models are motor neuron (MN)-intrinsic is unknown. We investigated whether human pluripotent stem cell-derived MNs form PNN-like structures in vitro, and whether ALS-associated mutations alter this process. We show that PNN-like structures containing hyaluronan, tenascin-R, and aggrecan form in in vitro co-cultures of iPSC-derived MNs and astrocytes, and that their formation and gene expression were not altered by ALS mutations. To explore whether PNN dysregulation reflects contributions from other cell types or selective MN vulnerability, we conducted meta-analyses of transcriptomic datasets from pluripotent stem cell-derived astrocytes carrying ALS-associated mutations, as well as datasets comparing MN populations with differential susceptibility to ALS. These analyses revealed no consistent differences in PNN-related gene expression within human stem cell-derived MNs. In contrast, transcriptomic analyses of human post-mortem ALS tissues revealed dysregulation of PNN-related genes, including core PNN components. Taken together, these findings indicate that PNN-related alterations described in ALS animal models are not reproduced by ALS-associated mutations in MNs alone, and instead point to a role for additional cellular components within the central nervous system.
    DOI:  https://doi.org/10.1038/s42003-026-10244-6
  10. J Cell Biol. 2026 Jul 06. pii: e202509040. [Epub ahead of print]225(7):
      The small GTP-binding protein Arl8b is established as a regulator of lysosome positioning and fusion, yet its role in lysosome biogenesis remains unclear. Here, we investigate the role of Arl8b in the trafficking of newly synthesized LAMP1 to lysosomes using the Retention Using Selective Hook (RUSH) assay. We find that Arl8b localizes to post-endocytic LAMP1-containing vesicles prior to fusion with acidic lysosomes. Arl8b depletion leads to Rab11a-dependent recycling of LAMP1 to the plasma membrane, impairing its lysosomal delivery. Mechanistically, Arl8b recruits the Rab11a GAP, TBC1D9B, to LAMP1-positive membranes, and TBC1D9B depletion similarly disrupts LAMP1 sorting. Notably, TBC1D9B knockdown also impairs the retrieval of cation-independent mannose-6-phosphate receptor (CI-M6PR) from Rab11a- and Rab14-positive endosomes to the trans-Golgi network, impairing pro-cathepsin trafficking and cargo degradation. These findings reveal that Arl8b-mediated recruitment of Rab GAP TBC1D9B is crucial for inactivation of the Rab11a recycling pathway, leading to efficient sorting of lysosomal cargo to their functional location.
    DOI:  https://doi.org/10.1083/jcb.202509040
  11. J Neurochem. 2026 May;170(5): e70465
      Understanding the mechanisms that underlie neuronal dynamics is crucial to promoting brain health. The chromatin-modifying protein RCOR2 (also named CoREST2) has recently gained attention for its neuroprotective roles. Although previous work has firmly established RCOR2's importance in neurogenesis and aging, its contributions to post-mitotic neurons remain understudied. This gap limits our ability to unveil its broader significance for brain health. In this study, we used the rat primary culture of embryonic hippocampal neurons, highly enriched in pyramidal glutamatergic neurons, to show that RCOR2 preserves the structural integrity of axons and dendrites. By combining state-of-the-art imaging techniques, including confocal, AiryScan2, and STED microscopy, we unveiled that RCOR2 is highly packed within neuronal nuclei, where it maintains the spatial organization of heterochromatin. The delivery of shRNA sequences targeting RCOR2, either by transient transfection or lentiviral infection, led to its partial knockdown and a pronounced shortening of axons and dendrites. This phenotype was paralleled by an abnormal accumulation of microtubule-associated proteins (MAPs), including MAP2 and Tau, revealed by qRT-PCR, immunoblotting, and confocal imaging. Strikingly, RCOR2 knockdown neurons show increased axonal MAP2, suggesting the loss of axonal identity. Moreover, z-stack imaging revealed an abnormal increase in the tyrosinated-tubulin/acetylated-tubulin ratio-a molecular marker of microtubule (MT) stability-indicating reduced MT stability. In this regard, the treatment with a nanomolar and MT-stabilizing dose of taxol (3 nM) partially rescued the neuritic growth of RCOR2 knock-down neurons, suggesting a MT-dependent mechanism. These findings unveil neuron-specific functions of RCOR2, highlighting its protective role in sustaining neuronal architecture during early development.
    Keywords:  CoREST2; MAP2; acetylation; chromatin; doublecortin; neurodegeneration; neuroprotection; tau; tubulin; tyrosination
    DOI:  https://doi.org/10.1111/jnc.70465
  12. J Alzheimers Dis. 2026 May 21. 13872877261452598
      BackgroundThe commonality of Alzheimer's disease (AD) in the elderly suggests connections between aging and AD biology. APOE biology is also tied to AD.ObjectiveWe sought to link three aging hallmarks (loss of proteostasis, mitochondrial dysfunction, deregulated nutrient sensing) to APOE biology.MethodsWe altered SH-SY5Y cell proteostasis directly via heat shock, integrated stress response inhibition (ISRIB), or autophagy inhibition (chloroquine), and indirectly by perturbing mitochondria (mtDNA depletion; oligomycin). We also exposed induced pluripotent stem cell-derived neurons to ISRIB and chloroquine. Conversely, we mitigated protein stress with rapamycin. We assessed intervention impact on APOE expression.ResultsIncreasing protein stress elevated and decreasing protein stress lowered APOE expression. In SH-SY5Y cells rapamycin blocked oligomycin-induced mTOR 2448 phosphorylation, Akt 473 phosphorylation, and APOE expression. In chloroquine-treated neurons rapamycin reduced mTOR phosphorylation and APOE expression.DiscussionProtein stress initiates APOE expression and facilitates mitochondrial dysfunction's impact on APOE by engaging the mTOR pathway. Our findings link aging and AD biology.
    Keywords:  APOE; Alzheimer's disease; mTOR; mitochondria; proteostasis; rapamycin
    DOI:  https://doi.org/10.1177/13872877261452598
  13. Cytoskeleton (Hoboken). 2026 May 18. e70145
      Proper regulation of microtubule (MT) cytoskeleton forms the basis for molecular and functional polarization of neurons. The optimal growth of mechanosensory neurons in C. elegans depends on the activity of the microtubule depolymerizing enzyme, Kinesin-13/KLP-7. Yet it's unclear how KLP-7 collaborates with other microtubule regulators in neurons for proper MT regulation. Using the ectopic growth of mechanosensory neurons in klp-7 mutants as a phenotypic handle, we characterized its genetic interactions with known microtubule stabilizers. We found that mutations in unc-33(CRMP-2), unc-44(Ankyrin), or components of the unc-14/unc-51/vab-8 pathway suppress klp-7(0) ectopic growth. Quantification of EBP-2::GFP dynamics revealed that these factors promote microtubule stability in axon-like PLM anterior neurites, suggesting an antagonistic role to KLP-7. Additionally, mec-7 (β-tubulin), unc-33, and unc-44 proteins promote the plus-end-out microtubule polarity in these neurites, and their loss in klp-7(0) mutant can reciprocally suppress the microtubule dynamics defects observed in single mutants. Notably, the simultaneous loss of klp-7 and mec-7 restored steady-state microtubule dynamics and wild-type-like PLM morphology. Together, our findings suggest that the antagonistic interplay between microtubule depolymerizing kinesin KLP-7 and microtubule-stabilizing factors, UNC-44, UNC-33, and tubulins, is necessary to maintain steady-state microtubule dynamics and plus-end-out axonal microtubule polarity in neurons.
    Keywords:   C. elegans ; KLP‐7; MEC‐7; UNC‐33; UNC‐44; UNC‐76; microtubules; touch neurons
    DOI:  https://doi.org/10.1002/cm.70145
  14. bioRxiv. 2026 May 07. pii: 2026.05.04.722784. [Epub ahead of print]
      In neurons, the endoplasmic reticulum (ER) forms an extensive network that establishes membrane contact sites (MCSs) with various organelles including the plasma membrane (PM). While MCSs are known to regulate lipid exchange and Ca 2+ signaling, their specific roles in synaptic transmission remain poorly understood. Here, we demonstrate that the ER resident proteins VAPA and VAPB are essential for organizing presynaptic Ca 2+ exchange and mobilizing synaptic vesicles. We show that the loss of VAP impairs Ca 2+ loading into both the ER and mitochondria during electrical activity. This regulation occurs primarily through VAP interactions with voltage-gated potassium channels (Kv2) at the PM. Our data suggest that the Kv2-VAP complex organizes presynaptic Ca 2+ signaling outside of the active zone. Without this scaffold, synaptic vesicles become trapped in the reserve pool and fail to participate in exocytosis. These findings reveal a novel role for Kv2-VAP MCSs in coordinating organelle Ca 2+ signaling and the synaptic vesicle cycle.
    DOI:  https://doi.org/10.64898/2026.05.04.722784
  15. EMBO Rep. 2026 May 22.
      Brain aging is an intricate process that inevitably leads to functional deterioration. However, its molecular drivers remain unclear. Here, we show that the age-related decline in LINC complex expression on the neuronal nuclear envelope impairs axon initial segment (AIS)-mediated excitability and triggers brain aging. With aging, the expression of LINC complex components, including Sun1, decreases in various brain regions, accompanied by a reduction in AIS length. Preserving Sun1 expression rescues nuclear structural abnormalities in aged neurons, shifting chromatin dynamics and global gene expression toward those of young neurons. Particularly, it restores the expression of AIS-related molecules, including voltage-gated sodium or potassium channels essential for action potential generation. Inhibiting the LINC complex in young mice impairs AIS integrity, leading to reduced neuronal excitability and brain dysfunction. Furthermore, Sun1 administration to aged neurons prevents age-related AIS shortening, excitability impairment, and brain function changes. Thus, we uncover the mechanism of normal brain aging involving AIS dysfunction, identifying the LINC complex component Sun1 as essential for preserving brain function.
    DOI:  https://doi.org/10.1038/s44319-026-00786-5
  16. medRxiv. 2026 May 04. pii: 2026.05.04.26352054. [Epub ahead of print]
      TDP-43 dysfunction is a defining feature of amyotrophic lateral sclerosis (ALS), yet no biofluid biomarker directly measures its functional activity. We developed a serum-based homogeneous time-resolved FRET (hTR-FRET) assay that quantifies TDP-43 RNA-binding activity using synthetic UU rich RNA probes. We analyzed 1,080 serum samples from controls, sporadic ALS, and genetic subgroups (C9orf72, SOD1) across multiple biorepositories. Cross-sectionally, TDP-43 ligation activity was elevated in ALS (mean 390 a.u.) versus controls (304 a.u.), yielding AUC = 0.79. Genotype means were 392 a.u. (sporadic), 382 a.u. (C9orf72), and 323 a.u. (SOD1); with a 366 a.u threshold achieved 95% specificity against controls. Longitudinally, Target ALS showed a modest but significant inverse correlation between TDP-43 activity and ALSFRS-R, while other cohorts exhibited similar non-significant trends. Elevated signal likely reflects increased extracellular, probe-competent TDP-43 species. This assay provides direct functional measurement of disease-relevant TDP-43 biology, supporting applications in diagnostic discrimination, genotype stratification, and progression monitoring in prospective studies.
    DOI:  https://doi.org/10.64898/2026.05.04.26352054
  17. Front Hum Neurosci. 2026 ;20 1768774
      Aging is a complex biological process characterized by progressive functional decline and increased vulnerability to age-related diseases, particularly neurodegenerative disorders. At the biological level, aging is characterized by a range of molecular and cellular mechanisms, including genomic instability, telomere attrition, loss of proteostasis, mitochondrial dysfunction, and chronic inflammation, which collectively contribute to cognitive decline and neuronal dysfunction over time. These hallmarks do not function independently but instead interact with one another during aging and neurodegeneration. Consequently, brain aging and neurodegenerative diseases are recognized as closely interconnected processes. To better understand this relationship, it is essential to examine the shared molecular and cellular mechanisms that link brain aging to neurodegeneration. In this review, we summarize the principal mechanisms underlying aging and neurodegenerative diseases, examine their roles in these processes, and highlight how their interactions shape both aging and neurodegeneration. We also discuss potential therapeutic strategies targeting key mechanisms involved in aging and neurodegeneration.
    Keywords:  brain aging; genomic instability; inflammation; loss of proteostasis; mitochondrial dysfunction; neurodegenerative diseases; telomere attrition; therapeutic strategies
    DOI:  https://doi.org/10.3389/fnhum.2026.1768774
  18. Mitochondrion. 2026 May 19. pii: S1567-7249(26)00061-9. [Epub ahead of print] 102171
      Myoclonic epilepsy with ragged-red fibers (MERRF) syndrome is mainly caused by the m.8344A > G mutation and mitochondrial dysfunction, but the pathogenesis remains unclear. In this study, we demonstrated that carbonyl cyanide m-chlorophenyl hydrazine (CCCP) induced PINK1-mediated mitophagy and accelerated mitochondrial turnover in the skin fibroblasts of MERRF patients. We found that CCCP led to more pronounced increase of PINK1 accumulation, activation of LC3B II and degradation of Mfn1, Mfn2, OSCP and OPA1 cleavage in MERRF skin fibroblasts as compared with normal skin fibroblasts. Moreover, N-acetylcysteine suppressed PINK1 accumulation and ubiquitin phosphorylation and thus impaired clearance of damaged mitochondria. This inhibitory effect was validated in MERRF patient iPSC-derived neurons harboring the m.8344A > G mutation, which displayed mitochondrial dysfunction, ROS overproduction and impaired electrophysiological function of mature neurons. These findings suggest that oxidative stress plays a crucial role in the susceptibility to mitophagy of skin fibroblasts and iPSC-derived neurons of MERRF patients and that restoring proper mitophagic flux is a potential therapeutic approach.
    Keywords:  MERRF syndrome; Mitophagy; N-acetylcysteine; PINK1; iPSC-derived neural stem cells (iNSCs); iPSC-derived neurons; mtDNA mutation
    DOI:  https://doi.org/10.1016/j.mito.2026.102171
  19. ACS Sens. 2026 May 21.
      Semiconductor quantum dots (QDs) provide unique stability, brightness, and multiplexed capacity for biomarker detection in complex diseases; however, their distinctive intracellular distribution has rarely been leveraged for spatially resolved diagnostics. Here, we show how QD-based sensors enable selective detection of cytoplasmic proteins and can quantify nucleo-cytoplasm protein mislocalization in patient-derived samples. We validated this approach labeling TAR DNA-binding protein 43 (TDP-43), a key mislocalized protein in amyotrophic lateral sclerosis (ALS). Spatial resolution is achieved in several patient-derived models and mouse brain tissue, underscoring the nanosensor's versatility across biological systems. Multiplexed QD-based immunolabeling, combined with confocal imaging and high-throughput flow cytometry, enables the detection of distinct cytoplasmic biomarker signatures that discriminate ALS patients from healthy controls. These signatures include variations in TDP-43 mislocalization and protein coexpression patterns, which were further modulated by pharmacological treatment. This work establishes QDs as spatially selective, multiplexable nanosensors capable of resolving subtle yet disease-relevant intracellular phenotypes in patient-derived samples. Compared to organic fluorophores, QDs enhance sensitivity, improve signal stability, and enable simultaneous spatially resolved biomarker quantification, broadening their potential for clinical diagnostics and personalized medicine. These findings establish QDs as powerful tools for neurodegeneration research, disease monitoring, and early biomarker discovery, with potential applications in translational neuroscience and precision medicine.
    Keywords:  TDP-43; amyotrophic lateral sclerosis; multiplexed protein detection; protein mislocalization; quantum dots
    DOI:  https://doi.org/10.1021/acssensors.5c01941
  20. Neuroscience. 2026 May 19. pii: S0306-4522(26)00333-7. [Epub ahead of print]
      Midbrain dopamine (DA) neurons are essential regulators of basal ganglia function. Their axonal structure is intricate, with numerous non-synaptic release sites and fewer synaptic terminals that notably release glutamate or GABA. Despite their significance, the molecular mechanisms governing DA neuron connectivity and neurochemical identity remain poorly understood. We hypothesize that trans-synaptic cell adhesion molecules such as neurexins (Nrxns) regulate the interactions of DA neuron axons with target cells and thereby influence axonal branching and synapse formation by DA neurons. We therefore examined neuronal survival, axonal growth and synapse formation in cultured DA neurons lacking all neurexins (DAT::NrxnsKO). Conditional deletion of all Nrxns in DA neurons revealed that loss of Nrxns does not disrupt the basic development of these neurons or the structure of their axonal terminals, including normal expression of the vesicular monoamine transporter (VMAT2) and the calcium sensor synaptotagmin 1 (Syt1). However, we observed a reduction in the number of TH-positive DA neurons in culture, suggesting altered resilience or increased vulnerability under in vitro conditions. In addition, loss of Nrxns selectively reduced the proportion of DA neuron terminals associated with the inhibitory postsynaptic marker gephyrin, while the association with excitatory synaptic markers was preserved.
    Keywords:  Active zone proteins; Dopamine; Neurexins; Synapses; Volume transmission
    DOI:  https://doi.org/10.1016/j.neuroscience.2026.05.022
  21. Mamm Genome. 2026 May 22. pii: 72. [Epub ahead of print]37(1):
      Charcot-Marie-Tooth disease (CMT) is a genetic peripheral neuropathy arising from mutations in diverse genes that principally disrupt axons and Schwann cells. As the most distal synaptic interface of motor neurons, the neuromuscular junction (NMJ) represents a plausible but underexplored site at which such disruptions may converge to confer selective peripheral neuropathy. This review synthesises current evidence for NMJ involvement in CMT, focusing on mammalian systems, and evaluates how localised synaptic pathology relates to distal nerve dysfunction across genetic models. We outline the organisation of the mammalian NMJ and experimental approaches used to assess its dysregulation, emphasising the distinction between structural and functional denervation. Appraisal of NMJ abnormalities reported across axonal and demyelinating CMT models reveals evidence for impaired synaptic maturation, transmission and conduction failure, often prior to subsequent structural denervation and axonal degeneration. Emerging patterns indicate well-studied axonal subtypes show early, length-dependent synaptic dysfunction, whereas demyelinating forms often exhibit secondary NMJ destabilisation with ineffective axonal sprouting and reinnervation attempts. We also address methodological and interpretive considerations in NMJ studies, and consider the translational relevance of NMJ disruption as a functional readout of pathology and potential therapeutic target. Collectively, this review clarifies the NMJ as an informative, active and selective site of vulnerability in CMT, while demonstrating both the need and relevance for additional investigation in mammalian systems.
    Keywords:  Axon degeneration; CMT; Motor neuron; Peripheral neuropathy; Schwann cell; Synapse
    DOI:  https://doi.org/10.1007/s00335-026-10238-z
  22. Mol Ther Adv. 2026 Jun 11. 34(2): 201741
      An intronic hexanucleotide repeat expansion in C9orf72 is the leading genetic cause of both frontotemporal dementia and amyotrophic lateral sclerosis (C9-FTD/ALS). We have previously demonstrated that CRISPR-Cas9 excision of the repeat expansion in patient iPSCs reverts pathological hallmarks of C9-FTD/ALS. Here, we aim to identify efficient and safe gRNAs for CRISPR-spCas9 dual-gRNA excision of the C9-repeat expansion. Utilizing novel ddPCR and single-molecule sequencing assays, we screened 120 gRNA pairs, comparing 64 bi-allelic, intronic excisions of the repeat region to 56 allele-specific excisions of the mutant allele in patient iPSCs, ranking them by efficiency. Bi-allelic excisions of the intronic repeat region were more efficient than excisions of the mutant allele. Single gRNA indel rates can nominate likely efficient gRNA pairs, but these pairs must be tested empirically. The length of the repeat expansion did not impact excision efficiency; rather, the activity of individual gRNAs drove excision efficiencies. Using whole genome sequencing and INDUCE-seq, we found only one detectable off-target of those nominated by Cas-OFFinder and CHANGE-seq across 4 of the most efficient gRNAs. This study advances the development of targeted therapies for C9-FTD/ALS and establishes a framework for dual-gRNA screening in patient iPSCs applicable to other repeat expansions.
    Keywords:  C9orf72; CRISPR; allele-specific; amyotrophic lateral sclerosis, ALS; arrayed CRISPR gRNA screen; dementia; dual-gRNA; frontotemporal dementia, FTD; gene therapy; iPSCs; motor-neuron disease; repeat expansion
    DOI:  https://doi.org/10.1016/j.omta.2026.201741
  23. JCI Insight. 2026 May 19. pii: e202204. [Epub ahead of print]
      Biallelic loss-of-function variants in the adaptor protein complex 4 (AP-4) disrupt trafficking of transmembrane proteins at the trans-Golgi network, including the autophagy-related protein 9A (ATG9A), leading to childhood-onset hereditary spastic paraplegia (AP-4-HSP). AP-4-HSP is characterized by features of both a neurodevelopmental and degenerative neurological disease. To investigate the molecular mechanisms underlying AP-4-HSP and identify potential therapeutic targets, we conducted an arrayed CRISPR/Cas9 loss-of-function screen of 8,478 genes, targeting the 'druggable genome', in a human neuronal model of AP-4 deficiency. Through this phenotypic screen and subsequent experiments, key modulators of ATG9A trafficking were identified, and complementary pathway analyses provided insights into the regulatory landscape of ATG9A transport. Knockdown of ANPEP and NPM1 enhanced ATG9A availability outside the trans-Golgi network, suggesting they regulate ATG9A localization. These findings deepen our understanding of ATG9A trafficking in the context of AP-4 deficiency and offer a framework for the development of targeted interventions for AP-4-HSP.
    Keywords:  Adaptor proteins; Cell biology; Genetic diseases; Genetics; Neurological disorders; Neuroscience
    DOI:  https://doi.org/10.1172/jci.insight.202204
  24. J Proteomics. 2026 May 19. pii: S1874-3919(26)00083-7. [Epub ahead of print]330 105680
      Human pluripotent stem cells can be differentiated into a variety of different cell types, for instance cardiomyocytes. Especially in the context of future application in regenerative medicine, it is crucial to understand the developmental processes taking place upon differentiation. Moreover, the identification of a panel of cell surface markers suitable for characterization and purification is necessary to ensure quality of human pluripotent stem-cell derived products. In this study, we used quantitative mass spectrometry to characterize proteomic changes in early mesendodermal differentiation. Two human pluripotent stem cell lines, one embryonic (H3) and one induced pluripotent stem cell line (I2), were analyzed under pluripotent conditions and after two days of embryoid body-based differentiation. Functional clustering and enrichment analysis showed down-regulation of proteins associated with pluripotency and the tricarboxylic acid cycle at day two. In contrast, proteins related to the proteasome and annexin family were up-regulated upon differentiation. Among the proteins that were down-regulated upon differentiation in both, H3 and I2, the membrane protein junctional adhesion molecule A (JAM-A) emerged as potentially associated with pluripotency. Flow cytometry and immunocytochemistry further confirmed down-regulation of JAM-A on the cell surface of human induced pluripotent stem cells that were differentiated toward mesendoderm for just two days. STATEMENT OF SIGNIFICANCE: Understanding early molecular changes during human pluripotent stem cell differentiation is essential for stem cell biology and regenerative medicine. This study provides a comparative proteomic analysis of two cell lines during early differentiation and identifies coordinated metabolic and pluripotency-associated changes. The understanding of changes that occur in the early phase of cardiomyocyte differentiation might be helpful to more precisely monitor the differentiation process. Importantly, we identify the membrane protein JAM-A as a robustly down-regulated cell surface protein, highlighting its potential as marker for human pluripotent stem cells. In the future, JAM-A might be used in a panel of cell surface markers for pluripotent stem cells in order to remove pluripotent stem cells from stem cell-derived therapeutic products.
    Keywords:  Cell surface marker; Embryoid body-based differentiation; Human pluripotent stem cells; JAM-A; Proteomics
    DOI:  https://doi.org/10.1016/j.jprot.2026.105680
  25. Eur J Med Chem. 2026 May 14. pii: S0223-5234(26)00396-X. [Epub ahead of print]315 118951
      TRPML1, a lysosomal Ca2+ channel, has emerged as a clinically relevant target due to its genetic and mechanistic links to lysosomal storage disorders and neurodegenerative diseases, including Gaucher disease, Parkinson's disease, Alzheimer's disease, and amyotrophic lateral sclerosis. This evidence has prompted TRPML1 drug discovery efforts across academia and industry, with several small-molecule agonists advancing toward clinical development. In this review, we provide a comprehensive overview of the therapeutic potential of TRPML1 as a molecular target from a medicinal chemistry perspective. We summarize the structural basis of channel activation and inhibition, highlighting insights from recent cryo-EM studies that define the principal ligand-binding sites and mechanisms of allosteric modulation. We systematically survey the chemical space of TRPML1 ligands reported to date, including diverse agonist and antagonist chemotypes, and extend this analysis to encompass undisclosed or recently disclosed compounds emerging from industry pipelines. Furthermore, we discuss key determinants of ligand design and developability, including the challenges associated with targeting a deeply embedded, lipophilic binding pocket within the membrane. Overall, the available evidence positions TRPML1 as a promising target for small-molecule drug discovery and provides a framework for the rational design of next-generation lysosome-directed therapeutics.
    Keywords:  Lysosomal dysfunction; Lysosomal storage disorders; Neurodegenerative diseases; Protein aggregates; TRPML1 agonist
    DOI:  https://doi.org/10.1016/j.ejmech.2026.118951
  26. Cell Death Dis. 2026 May 19.
      Autism spectrum disorder (ASD) presents a major clinical challenge, necessitating the identification of novel therapeutic targets rooted in its underlying pathophysiology. The axon initial segment (AIS) is the critical site for action potential initiation and a hub for homeostatic plasticity; however, its involvement in ASD remains poorly defined. Herein, we report significant structural and functional deficits in the AIS within a clinically relevant ASD mouse model harboring a 15q11-13 duplication (15q dup). We observed that pyramidal neurons in the medial prefrontal cortex (mPFC) exhibited shortened AIS, resulting in reduced neuronal excitability and impaired plasticity. Importantly, these abnormalities were specific to long-range circuits, including the mPFC-dorsal raphe nucleus (DRN) pathway, which is critical for social behavior. We employed a circuit-specific chemogenetic strategy that activates these mPFC-DRN projection neurons to test the reversibility of this phenotype. Remarkably, this targeted intervention normalized AIS structure and rescued core ASD-like behaviors, including social interaction deficits and repetitive behaviors. These results demonstrated that AIS alterations in this ASD model represent a reversible form of maladaptive plasticity, rather than permanent neuropathology. Our study highlights circuit-specific AIS modulation as a promising novel avenue for therapeutic interventions aimed at correcting fundamental neuronal excitability deficits in ASD.
    DOI:  https://doi.org/10.1038/s41419-026-08873-0
  27. Neuroscience. 2026 May 20. pii: S0306-4522(26)00341-6. [Epub ahead of print]
      TAR DNA-binding protein 43 (TDP-43) proteinopathy has recently emerged as a pivotal, yet underrecognized, contributor to the multifaceted neuropathology of Alzheimer's disease (AD). While amyloid-β and tau have long been established as cardinal pathological hallmarks, growing evidence delineates TDP-43 as a critical participant of neurodegeneration, intricately interwoven with amyloid and tau pathologies. TDP-43 mislocalization, post-translational modifications, and aggregation potentiate neuronal loss through disruption of RNA metabolism, nucleocytoplasmic transport, and protein homeostasis. This tripartite interplay manifests in synergistic and possibly multidirectional pathological cascades that amplify neuronal vulnerability and cognitive decline, thereby complicating the clinical and pathological complexity of AD. Here, we critically reviewed the mechanistic crosstalk among TDP-43, amyloid-β, and tau, focusing on preclinical and clinical evidence, highlighting possible convergent pathways of aggregation, propagation, and neurodegeneration. Moreover, this review also evaluates mitochondrial dysfunction, autophagy failure, and inflammation as underlying events associated with TDP-43 pathology. Therefore, we argue for a reconceptualization of AD as a dynamic proteinopathy network, with TDP-43 as a core integrative node influencing disease onset and its progression. Notably, we discuss emerging diagnostic modalities associated with molecular tracers of TDP-43, providing prospects for future biomarker identification. Finally, this review articulates the translational relevance of TDP-43 therapy in AD and related neurological disorders, emphasizing the necessity of holistic approaches that transcend the traditional amyloid-tau paradigm to effectively tackle the full spectrum of AD pathobiology.
    Keywords:  Amyloid beta; Autophagy; Inflammation; Mitochondrial dysfunction; Protein Aggregates; Tau
    DOI:  https://doi.org/10.1016/j.neuroscience.2026.05.024
  28. bioRxiv. 2026 May 05. pii: 2026.04.30.721979. [Epub ahead of print]
      Three-dimensional stem cell-derived neural organoids provide a promising platform for investigating early brain development and interregional circuit formation. Although co-culture of region-specific organoids into assembloids has enabled the study of cortical and subcortical interactions, these models lack directional specificity and spatial control, limiting their ability to recapitulate canonical circuit architecture. Here, we present a microfluidic platform for constructing directional and tunable interregional circuits while preserving anatomical distinction. This system, which we term "directoids" incorporates micropatterned polydimethylsiloxane (PDMS) microstructures to control uni- and bidirectional axonal growth between cortical and thalamic organoids. We observed a 70.4% success rate of axons traversing the full channel length in the permissive direction and reaching the opposing organoid, whereas no neurites successfully crossed the probative direction. These results demonstrate robust directionally bias in axon outgrowth and establish a scalable, reproducible strategy for controlling asymmetric connectivity between anatomically distinct neural organoids. Using high-density CMOS microelectrode arrays, we further validated directional tuning of extracellular action potential propagation within directoid microchannels, a feature not observed in straight-channel connectoid controls. Directoids also exhibited significant asymmetry in firing rates between channel entry and exit sites, consistent with engineered bias in signal flow. This provides an experimental paradigm for dissecting how anatomical connectivity and functional activity converge to shape neuronal networks. Together, these findings establish a microfluidic platform for investigating the mechanisms underlying hierarchical circuit formation, regional specification, and functional integration in developing human neural organoid models at cellular resolution not possible in vivo .
    DOI:  https://doi.org/10.64898/2026.04.30.721979
  29. Neurochem Res. 2026 May 22. pii: 169. [Epub ahead of print]51(3):
      Transactive Response Deoxyribonucleic Acid-Binding Protein-43 (TDP-43) acetylation may couple motor-neuron degeneration to systemic immune orchestration in Amyotrophic Lateral Sclerosis (ALS). Upon nuclear clearance and mislocalisation, TDP-43 enters the periphery; acetylation shapes its conformation, trafficking and immunogenicity. This narrative review synthesises single-cell transcriptomics, proteomic immunoprofiling and clinical inflammatory phenotyping to examine whether site-specific acetylated TDP-43 species may be associated with peripheral inflammatory signatures relevant to ALS immunopathology. By integrating separate datasets on acetylated TDP-43, monocyte phenotypes and cytokine modules, we propose two provisional endotypes characterised by monocyte reprogramming, cytokine modules and Blood-Brain Barrier (BBB) dysfunction-each representing clinically actionable pathways. Framed as a provisional neuroimmune interface, the acetylation state is considered here as a plausible molecular correlate and potential therapeutic entry point: a measurable clue to inform pharmacological targeting and, potentially, a modifiable target via p300CREB-Binding Protein (CBP)-Histone Deacetylase (HDAC) axes or sirtuin activity. Recasting TDP-43 from neuropathological hallmark to immunoactive sentinel supports a shift from descriptive nosology to stratified immunotherapy, in which treatment allocation is informed by acetylation-defined peripheral signatures.
    Keywords:  Amyotrophic Lateral Sclerosis; Immune stratotype; Peripheral neuroinflammation; Post-translational modification; TDP-43 acetylation
    DOI:  https://doi.org/10.1007/s11064-026-04770-2
  30. Sci Adv. 2026 May 22. 12(21): eaeb8658
      In p53-deficient cancers, targeting cholesterol metabolism has emerged as a promising therapeutic approach, given that p53 loss dysregulates sterol regulatory element-binding protein 2 pathways, thereby enhancing cholesterol biosynthesis. While cholesterol synthesis inhibitors such as statins have shown initial success, their efficacy is often compromised by the development of acquired resistance. Consequently, strategies are being explored to disrupt cholesterol homeostasis more comprehensively by inhibiting its synthesis and intracellular transport. In this study, we investigate a previously underexplored function of PI5P4Ks, which catalyzes the conversion of PI(5)P to PI(4,5)P2 at intracellular membranes. Our findings reveal that PI5P4Ks play a key role in facilitating lysosomal cholesterol transport, regulating lysosome positioning, and sustaining growth signaling via the mechanistic target of rapamycin (mTOR) pathway. While PI5P4Ks have previously been implicated in mTOR signaling and tumor proliferation in p53-deficient contexts, this work elucidates an upstream mechanism that unifies these earlier observations.
    DOI:  https://doi.org/10.1126/sciadv.aeb8658
  31. Stud Health Technol Inform. 2026 May 21. 336 1705-1709
      Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder characterized by progressive motor neuron degeneration and limited diagnostic biomarkers. Identifying robust molecular biomarkers for ALS remains a major challenge due to disease heterogeneity and high-dimensional gene expression data. In this study, we developed a machine learning (ML) based pipeline integrating transcriptome data and feature selection to identify potential ALS biomarkers. RNA-Seq data of motor neuron disease patients and healthy controls were obtained from publicly available GEO datasets, followed by preprocessing was performed. We implemented two ensembled ML models such as eXtreme gradient boosting (XGBoost) and random forest (RF) algorithms under a five-fold stratified cross-validation framework to identify the differentially expressed genes. These models were evaluated using the performance metrics. We identified top 10 genes ranked by feature importance from the XGBoost and RF models. Notably, the DCN (Decorin) gene appears consistently in the top 10 features of both models, underscoring its stability and biological relevance. Both ML models exhibited excellent classification performance, with RF achieving 98.8% accuracy and XGBoost achieving 97.6% accuracy, alongside consistently high sensitivity, specificity, precision, and F1-score values. This work highlights the utility of transcriptomic data and ML in identifying key genes as biomarkers for diagnostic and therapeutic potential in ALS.
    Keywords:  Amyotrophic Lateral Sclerosis; Biomarkers; Feature Importance; Machine learning; Transcriptomic data
    DOI:  https://doi.org/10.3233/SHTI260516