bims-axbals Biomed News
on Axonal biology and ALS
Issue of 2025–05–04
fifty papers selected by
TJ Krzystek
  1. A guide to selecting high-performing antibodies for S1PR1 (UniProt ID: P21453) for use in western blot, immunoprecipitation, and immunofluorescence.
  2. High Content Analysis of Mitochondrial Function in Induced Pluripotent Stem Cell-Derived Neurons.
  3. Targeting TDP-43 Proteinopathy in hiPSC-Derived Mutated hNPCs with Mitoxantrone Drugs and miRNAs.
  4. Inborn errors of canonical autophagy in neurodegenerative diseases.
  5. The motor neuron m6A repertoire governs neuronal homeostasis and FTO inhibition mitigates ALS symptom manifestation.
  6. Co-cultures of Human-Induced Pluripotent Stem Cell-Derived Neurons, Astrocytes, and Microglia for Modeling Neurodegenerative Diseases.
  7. Truncation mutation of CHMP2B disrupts late endosome function but reduces TDP-43 aggregation through HSP70 upregulation.
  8. Role of glial cells in motor neuron degeneration in hereditary spastic paraplegias.
  9. Context-dependent Interactors Regulate TDP-43 Dysfunction in ALS/FTLD.
  10. The Molecular Intersection of NEK1, C21ORF2, Cyclin F, and VCP in ALS Pathogenesis.
  11. Using Human iPSC-Derived Astrocytes to Investigate Transcription Factor-Driven Astrocyte to Neuron Transdifferentiation.
  12. Metabolic dysregulation in amyotrophic lateral sclerosis: insights from 1H NMR-based metabolomics in a tertiary care center in India.
  13. Cross-tissue MiRNA profiling of extracellular vesicles and PBMCs from amyotrophic lateral sclerosis patients.
  14. TDP-43 seeding activity in the olfactory mucosa of patients with amyotrophic lateral sclerosis.
  15. The role of alpha-synuclein in synucleinopathy: Impact on lipid regulation at mitochondria-ER membranes.
  16. Widespread distribution of α-synuclein oligomers in LRRK2-related Parkinson's disease.
  17. Novel extracellular vesicle release pathway facilitated by toxic superoxide dismutase 1 oligomers.
  18. Genome-wide association analysis identifies APOE as a mitophagy modifier in Lewy body disease.
  19. Nanoscale domains govern local diffusion and aging within FUS condensates.
  20. Profiling the LAM Family of Contact Site Tethers Provides Insights into Their Regulation and Function.
  21. The potential of brain organoids in addressing the heterogeneity of synucleinopathies.
  22. Preclinical evaluation of AAV9-coSMN1 gene therapy for spinal muscular atrophy: efficacy and safety in mouse models and non-human primates.
  23. Image-based temporal profiling of autophagy-related phenotypes.
  24. Glutamate gradually elevates [Zn2+]i via the CaM-CaMKII-NOS cascade in primary cultured rat embryonic cortical neurons.
  25. Dysfunctional Mitochondria Characterize Amyotrophic Lateral Sclerosis Patients' Cells Carrying the p.G376D TARDBP Pathogenetic Substitution.
  26. Immunological Fluid Biomarkers in Frontotemporal Dementia: A Systematic Review.
  27. Deciphering the Inhibitory Mechanism of ALS-Associated N352S and S352p Variants against TDP-43 Aggregation and Its Destabilization Effect on TDP-43 Protofibrils.
  28. Mitochondria-Plasma Membrane Contact Sites: Emerging Regulators of Mitochondrial Form and Function.
  29. The Role of mTOR in Amyotrophic Lateral Sclerosis.
  30. A Novel Phosphorylated Tau Conformer Implicated in the Tauopathy Pathogenesis of Human Neurons.
  31. Challenges of modelling TDP-43 pathology in mice.
  32. Application of a high-density microelectrode array assay using a 3D human iPSC-derived brain microphysiological system model for in vitro neurotoxicity screening of environmental compounds.
  33. Differences in neuronal ciliation rate and ciliary content revealed by systematic imaging-based analysis of hiPSC-derived models across protocols.
  34. Elucidating the Synergistic Effect of the PrimeC Combination for Amyotrophic Lateral Sclerosis in Human Induced Pluripotent Stem Cell-Derived Motor Neurons and Mouse Models.
  35. VGF and Its Derived Peptides in Amyotrophic Lateral Sclerosis.
  36. Skeletal muscle biomarkers of amyotrophic lateral sclerosis: a large-scale, multi-cohort proteomic study.
  37. Emerging roles of transfer RNA fragments in the CNS.
  38. Attenuated PINK1 autophosphorylation play neuroprotective and anti-seizure roles in neonatal hypoxia.
  39. Organoid Vascularization: Strategies and Applications.
  40. Antisense Oligonucleotides: A Promising Advancement in Neurodegenerative Disease Treatment.
  41. Purification of Low-Complexity Domain Proteins FUS, EWSR1, and Their Fusions.
  42. Oligonucleotide therapeutics for neurodegenerative diseases.
  43. Engineered Hydrogels for 3D Cell Culture and Bioprinting of Human Induced Pluripotent Stem Cell-Derived Neuroepithelial Stem Cells.
  44. A comparison of p-tau assays for the specificity to detect tau changes in Alzheimer's disease.
  45. Cell-Based Assays Using Derived Human-Induced Pluripotent Cells in Drug Discovery and Development.
  46. SIRTUINS MODULATORS MITIGATE HYPOXIA-INDUCED CELL DEATH DUE TO CHANGES IN HISTONE 3 ACETYLATION, AND MITOCHONDRIAL FUNCTION, DYNAMICS, AND CONTENT.
  47. A Novel Approach to Relocate Misplaced Proteins in Cells.
  48. 3D Mitochondrial Structure in Aging Human Skeletal Muscle: Insights Into MFN-2-Mediated Changes.
  49. A good side of STING.
  50. Extracellular matrix mechanical cues (dys)regulate metabolic redox homeostasis due to impaired autophagic flux.
  1. F1000Res. 2024 ;13 792
    A guide to selecting high-performing antibodies for S1PR1 (UniProt ID: P21453) for use in western blot, immunoprecipitation, and immunofluorescence.
    NeuroSGC/YCharOS/EDDU collaborative group
      Sphingosine 1-phosphate receptor 1 (S1PR1) is a G-coupled protein receptor that induces crucial biological processes when bound by sphingosine 1-phosphate. Here, we have characterized nine S1PR1 commercial antibodies for western blot, immunoprecipitation, and immunofluorescence using a standardized experimental protocol based on comparing read-outs in knockout cell lines and isogenic parental controls. These studies are part of a larger, collaborative initiative seeking to address antibody reproducibility issues by characterizing commercially available antibodies for human proteins and publishing the results openly as a resource for the scientific community. While use of antibodies and protocols vary between laboratories, we encourage readers to use this report as a guide to select the most appropriate antibodies for their specific needs.
    Keywords:  EDG1; S1PR1; UniProt ID P21453; antibody characterization; antibody validation; immunofluorescence; immunoprecipitation; sphingosine 1-phosphate receptor 1; western blot
    DOI:  https://doi.org/10.12688/f1000research.153244.2
  2. Methods Mol Biol. 2025 ;2924 259-268
    High Content Analysis of Mitochondrial Function in Induced Pluripotent Stem Cell-Derived Neurons.
    Daniel Little, Christin Luft, Olukunbi Mosaku, Robin Ketteler, Michael J Devine, Paul Gissen.
      Mitochondrial dysfunction is linked to many neurological diseases; therefore, the ability to measure mitochondrial function is of great use for researching disease and testing potential therapeutics. Here we describe a high-content assay to simultaneously measure mitochondrial membrane potential, morphology, and cell viability in iPSC-derived neurons. Neurons are seeded into plates suitable for fluorescent microscopy, and stained with the mitochondrial membrane potential-dependent dye TMRM, cytoplasmic dye Calcein-AM, and nuclear stain Hoechst-33,342. Images are acquired in live cells and analyzed using automated image analysis software.
    Keywords:  High content screening; Image analysis; Induced pluripotent stem cells; Mitochondria; Neurons
    DOI:  https://doi.org/10.1007/978-1-0716-4530-7_19
  3. Pharmaceutics. 2025 Mar 25. pii: 410. [Epub ahead of print]17(4):
    Targeting TDP-43 Proteinopathy in hiPSC-Derived Mutated hNPCs with Mitoxantrone Drugs and miRNAs.
    Uzair A Ansari, Ankita Srivastava, Ankur K Srivastava, Abhishek Pandeya, Pankhi Vatsa, Renu Negi, Akash Singh, Aditya B Pant.
      Background/Objectives: TDP-43 mutation-driven Amyotrophic Lateral Sclerosis (ALS) motor neuron disease is one of the most prominent forms (approximately 97%) in cases of sporadic ALS. Dysfunctional autophagy and lysosomal function are the prime mechanisms behind ALS. Mitoxantrone (Mito), a synthetic doxorubicin analog, is an inhibitor of DNA and RNA synthesis/repair via intercalating with nitrogenous bases and inhibiting topoisomerase II. The therapeutic potential of miRNAs associated with disease conditions has also been reported. This study explores the therapeutic potential of Mito along with miRNAs against mutated TDP-43 protein-induced proteinopathy in human-induced pluripotent stem cell (hiPSC)-derived human neural progenitor cells (hNPCs). Methods: HiPSCs mutated for TDP-43 were differentiated into hNPCs and used to explore the therapeutic potential of Mito at a concentration of 1 μM for 24 h (the identified non-cytotoxic dose). The therapeutic effects of Mito on miRNA expression and various cellular parameters such as mitochondrial dynamics, autophagy, and stress granules were assessed using the high-throughput Open Array technique, immunocytochemistry, flow cytometry, immunoblotting, and mitochondrial bioenergetic assay. Results: Mutated TDP-43 protein accumulation causes stress granule formation (G3BP1), mitochondrial bioenergetic dysfunction, SOD1 accumulation, hyperactivated autophagy, and ER stress in hNPCs. The mutated hNPCs also show dysregulation in six miRNAs (miR-543, miR-34a, miR-200c, miR-22, miR-29b, and miR-29c) in mutated hNPCs. A significant restoration of TDP-43 mutation-induced alterations could be witnessed upon the exposure of mutated hNPCs to Mito. Conclusions: Our study indicates that miR-543, miR-29b, miR-22, miR-200c, and miR-34a have antisense therapeutic potential alone and in combination with Mitoxantrone.
    Keywords:  ALS model; TDP-43; hiPSCs; high-throughput screening; human neural progenitor cells (hNPCs); miRNA; mitoxantrone
    DOI:  https://doi.org/10.3390/pharmaceutics17040410
  4. Hum Mol Genet. 2025 Apr 30. pii: ddae179. [Epub ahead of print]
    Inborn errors of canonical autophagy in neurodegenerative diseases.
    Dennis Freisem, Helene Hoenigsperger, Alberto Catanese, Konstantin M J Sparrer.
      Neurodegenerative disorders (NDDs), characterized by a progressive loss of neurons and cognitive function, are a severe burden to human health and mental fitness worldwide. A hallmark of NDDs such as Alzheimer's disease, Huntington's disease, Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS) and prion diseases is disturbed cellular proteostasis, resulting in pathogenic deposition of aggregated protein species. Autophagy is a major cellular process maintaining proteostasis and integral to innate immune defenses that mediates lysosomal protein turnover. Defects in autophagy are thus frequently associated with NDDs. In this review, we discuss the interplay between NDDs associated proteins and autophagy and provide an overview over recent discoveries in inborn errors in canonical autophagy proteins that are associated with NDDs. While mutations in autophagy receptors seems to be associated mainly with the development of ALS, errors in mitophagy are mainly found to promote PD. Finally, we argue whether autophagy may impact progress and onset of the disease, as well as the potential of targeting autophagy as a therapeutic approach. Concludingly, understanding disorders due to inborn errors in autophagy-"autophagopathies"-will help to unravel underlying NDD pathomechanisms and provide unique insights into the neuroprotective role of autophagy, thus potentially paving the way for novel therapeutic interventions.
    Keywords:  autophagy; innate immunity; monogenic diseases; neurodegenerative diseases
    DOI:  https://doi.org/10.1093/hmg/ddae179
  5. Nat Commun. 2025 Apr 30. 16(1): 4063
    The motor neuron m6A repertoire governs neuronal homeostasis and FTO inhibition mitigates ALS symptom manifestation.
    Ya-Ping Yen, Ting-Hsiang Lung, Ee Shan Liau, Chuan-Che Wu, Guan-Lin Huang, Fang-Yu Hsu, Mien Chang, Zheng-Dao Yang, Chia-Yi Huang, Zhong Zheng, Wei Zhao, Jui-Hung Hung, Chuan He, Qing Nie, Jun-An Chen.
      Amyotrophic lateral sclerosis (ALS) is a swiftly progressive and fatal neurodegenerative ailment marked by the degenerative motor neurons (MNs). Why MNs are specifically susceptible in predominantly sporadic cases remains enigmatic. Here, we demonstrated N6-methyladenosine (m6A), an RNA modification catalyzed by the METTL3/METTL14 methyltransferase complex, as a pivotal contributor to ALS pathogenesis. By conditional knockout Mettl14 in murine MNs, we recapitulate almost the full spectrum of ALS disease characteristics. Mechanistically, pervasive m6A hypomethylation triggers dysregulated expression of high-risk genes associated with ALS and an unforeseen reduction of chromatin accessibility in MNs. Additionally, we observed diminished m6A levels in induced pluripotent stem cell derived MNs (iPSC~MNs) from familial and sporadic ALS patients. Restoring m6A equilibrium via a small molecule or gene therapy significantly preserves MNs from degeneration and mitigates motor impairments in ALS iPSC~MNs and murine models. Our study presents a substantial stride towards identifying pioneering efficacious ALS therapies via RNA modifications.
    DOI:  https://doi.org/10.1038/s41467-025-59117-2
  6. Methods Mol Biol. 2025 ;2924 75-91
    Co-cultures of Human-Induced Pluripotent Stem Cell-Derived Neurons, Astrocytes, and Microglia for Modeling Neurodegenerative Diseases.
    Julian Roewe, Yasmin Riesinger, Ruven Wilkens, Heyne Lee, Daniela Geist, Tanja Georgi, Michael Bahr, Carina Habich, Lamiaa Bahnassawy, Nathalie Nicolaisen, Venus Thomanetz, Thomas Wassmer, Miroslav Cik, Peter Reinhardt.
      Although progressive neurodegenerative diseases like Alzheimer's disease (AD) have been extensively studied for decades, some underlying mechanisms of pathogenesis remain elusive. In addition, modeling neurodegenerative diseases in vitro has proven to be a challenging task. However, advances in the technique of using human induced pluripotent stem cells (hiPSCs) have enabled the scientific community to study hiPSC-derived neurons, astrocytes, or microglia. Despite this important progress, monocultures of individual cell types may not accurately reflect the complexities of modeling specific pathological mechanisms. Therefore, we present a robust protocol for co-cultivating hiPSC-derived cortical neurons, astrocytes, and microglia in a tri-culture system for the purpose of hypothesis testing and drug screening. This co-cultivation system may allow modeling the effect of astrocyte and/or microglia modulation on neuronal health or intra-neuronal Tau aggregation, a key feature of AD progression.
    Keywords:  Astrocytes; Microglia; Neurons; hiPSC
    DOI:  https://doi.org/10.1007/978-1-0716-4530-7_6
  7. Neurochem Int. 2025 Apr 30. pii: S0197-0186(25)00055-5. [Epub ahead of print] 105982
    Truncation mutation of CHMP2B disrupts late endosome function but reduces TDP-43 aggregation through HSP70 upregulation.
    Yohei Iguchi, Yuhei Takahashi, Jiayi Li, Yoshinobu Amakusa, Yu Kawakami, Takashi Yoshimura, Ryo Chikuchi, Madoka Iida, Satoshi Yokoi, Masahisa Katsuno.
      TAR DNA-binding protein 43 (TDP-43)-positive cytoplasmic aggregation is a pathological hallmark of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). This aggregation contributes substantially to the neurodegeneration of ALS and FTLD. The endosome, a key component of membrane trafficking in eukaryotic cells and is involved in the autophagy-lysosome pathway. Endosome-related genes such as CHMP2B, Alsin, and TMEM106B, are either causative or act as genetic modifiers in ALS and FTLD. However, the association between endosomal functions and TDP-43 aggregations remain poorly understood. The C-terminal truncation mutation CHMP2B, which causes frontotemporal dementia associated with chromosome 3 (FTD3), disrupts late endosome (LE)-lysosomes fusion. Nevertheless, FTD3 does not induce TDP-43 pathology. In this study, we showed that CHMP2B mutation-induced LE dysfunction promotes TDP-43 aggregate degradation through enhanced recruitment to juxtanuclear quality control compartments. Transcriptomic analysis revealed that CHMP2Bintron5 overexpression upregulates HSP70 expression. New insights into the connection between CMHP2B and HSP70 as well as the role of HSP70-mediated membrane trafficking in TDP-43 aggregation, offer a valuable understanding of the disease mechanism of ALS and FTLD.
    Keywords:  ALS; CHMP2B; FTD; TDP-43; endosome; juxtanuclear quality control compartment
    DOI:  https://doi.org/10.1016/j.neuint.2025.105982
  8. Front Cell Neurosci. 2025 ;19 1553658
    Role of glial cells in motor neuron degeneration in hereditary spastic paraplegias.
    Manaswini Vijayaraghavan, Sarvika Periyapalayam Murali, Gitika Thakur, Xue-Jun Li.
      This review provides a comprehensive overview of hereditary spastic paraplegias (HSPs) and summarizes the recent progress on the role of glial cells in the pathogenesis of HSPs. HSPs are a heterogeneous group of neurogenetic diseases characterized by axonal degeneration of cortical motor neurons, leading to muscle weakness and atrophy. Though the contribution of glial cells, especially astrocytes, to the progression of other motor neuron diseases like amyotrophic lateral sclerosis (ALS) is well documented, the role of glial cells and the interaction between neurons and astrocytes in HSP remained unknown until recently. Using human pluripotent stem cell-based models of HSPs, a study reported impaired lipid metabolisms and reduced size of lipid droplets in HSP astrocytes. Moreover, targeting lipid dysfunction in astrocytes rescues axonal degeneration of HSP cortical neurons, demonstrating a non-cell-autonomous mechanism in axonal deficits of HSP neurons. In addition to astrocytes, recent studies revealed dysfunctions in HSP patient pluripotent stem cell-derived microglial cells. Increased microgliosis and pro-inflammation factors were also observed in HSP patients' samples, pointing to an exciting role of innate immunity and microglia in HSP. Building upon these recent studies, further investigation of the detailed molecular mechanism and the interplay between glial cell dysfunction and neuronal degeneration in HSP by combining human stem cell models, animal models, and patient samples will open avenues for identifying new therapeutic targets and strategies for HSP.
    Keywords:  astrocytes; axonal degeneration; lipid dysfunction; microglia; motor neurons; neuroinflammation
    DOI:  https://doi.org/10.3389/fncel.2025.1553658
  9. bioRxiv. 2025 Apr 09. pii: 2025.04.07.646890. [Epub ahead of print]
    Context-dependent Interactors Regulate TDP-43 Dysfunction in ALS/FTLD.
    Longxin Xie, Yuehua Zhu, Bryan T Hurtle, Matthew Wright, John L Robinson, Jocelyn C Mauna, Emily E Brown, Marilyn Ngo, Cristian A Bergmann, Jiazhen Xu, Jessica Merjane, Amanda M Gleixner, Gabriela Grigorean, Feilin Liu, Wilfried Rossoll, Edward B Lee, Evangelos Kiskinis, Maria Chikina, Christopher J Donnelly.
      TDP-43 mislocalization, aggregation, and loss of splicing function are neuropathological hallmarks in over 97% of Amyotrophic Lateral Sclerosis (ALS), 45% of Frontotemporal Lobar Degeneration (FTLD), and 60% of Alzheimer's Disease, which has been reclassified as LATE-NC. However, the mechanisms underlying TDP-43 dysfunction remain elusive. Here, we utilize APEX2-driven proximity labeling and mass spectrometry to characterize the context-dependent TDP-43 interactome in conditions of cytoplasmic mislocalization, impaired RNA-binding contributing to aggregation, and oxidative stress. We describe context-dependent interactors, including disrupted interactions with splicing-related proteins and altered biomolecular condensate (BMC) associations. By integrating ALS and FTLD snRNA-seq data, we uncover disease-relevant molecular alterations and validate our dataset through a functional screen that identifies key TDP- 43 regulators. We demonstrate that disrupting nuclear speckle integrity, particularly through the downregulation of the splicing factor SRRM2, promotes TDP-43 mislocalization and loss of function. Additionally, we identify NUFIP2 as an interactor associated with mislocalization that sequesters TDP-43 into cytoplasmic aggregates and co-localizes with TDP-43 pathology in patient tissue. We also highlight HNRNPC as a potent TDP-43 splicing regulator, where precise modulation of TDP-43 or HNRNPC can rescue cryptic exon splicing. These findings provide mechanistic insights and potential therapeutic targets for TDP-43 dysfunction.
    DOI:  https://doi.org/10.1101/2025.04.07.646890
  10. Genes (Basel). 2025 Mar 30. pii: 407. [Epub ahead of print]16(4):
    The Molecular Intersection of NEK1, C21ORF2, Cyclin F, and VCP in ALS Pathogenesis.
    Yasuaki Watanabe, Tadashi Nakagawa, Makiko Nakagawa, Keiko Nakayama.
      Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disorder characterized by the progressive degeneration of motor neurons, leading to muscle weakness, paralysis, and death. Although significant progress has been made in understanding ALS, its molecular mechanisms remain complex and multifactorial. This review explores the potential convergent mechanisms underlying ALS pathogenesis, focusing on the roles of key proteins including NEK1, C21ORF2, cyclin F, VCP, and TDP-43. Recent studies suggest that mutations in C21ORF2 lead to the stabilization of NEK1, while cyclin F mutations activate VCP, resulting in TDP-43 aggregation. TDP-43 aggregation, a hallmark of ALS, impairs RNA processing and protein transport, both of which are essential for neuronal function. Furthermore, TDP-43 has emerged as a key player in DNA damage repair, translocating to DNA damage sites and recruiting repair proteins. Given that NEK1, VCP, and cyclin F are also involved in DNA repair, this review examines how these proteins may intersect to disrupt DNA damage repair mechanisms, contributing to ALS progression. Impaired DNA repair and protein homeostasis are suggested to be central downstream mechanisms in ALS pathogenesis. Ultimately, understanding the interplay between these pathways could offer novel insights into ALS and provide potential therapeutic targets. This review aims to highlight the emerging connections between protein aggregation, DNA damage repair, and cellular dysfunction in ALS, fostering a deeper understanding of its molecular basis and potential avenues for intervention.
    Keywords:  C21ORF2; DNA damage repair; NEK1; TDP-43 aggregation; VCP; amyotrophic lateral sclerosis (ALS); cyclin F; neurodegeneration; protein homeostasis
    DOI:  https://doi.org/10.3390/genes16040407
  11. Methods Mol Biol. 2025 ;2924 17-29
    Using Human iPSC-Derived Astrocytes to Investigate Transcription Factor-Driven Astrocyte to Neuron Transdifferentiation.
    Ruven Wilkens, Jürgen Korffmann, Nathalie Nicolaisen, Tjasa Lepko Modic, Julian Roewe, Carina Habich, Lamiaa Bahnassawy, Miroslav Cik, Peter Reinhardt.
      Neurodegenerative diseases such as Alzheimer's or Parkinson's are marked by progressive loss of affected neurons. Even with novel disease-modifying therapies, this loss cannot be reversed. In situ astrocyte-to-neuron (AtN) transdifferentiation may provide an opportunity to convert resident astrocytes into new neurons to revert the loss of neurons incurred. Currently, most studies investigating AtN transdifferentiation in vitro rely on the use of primary mouse astrocyte cultures which require sacrificing animals and come with uncertainty regarding species differences. Conversely, human induced pluripotent stem cell (hiPSC)-derived astrocytes offer the advantage of working in a human cell culture system which improves translatability and provides the opportunity to generate large, cryopreservable batches of cells to identify and study conversion factors. This protocol details a workflow for assessing the suitability of potential conversion factors for transdifferentiating hiPSC-derived astrocytes into neurons.
    Keywords:  Astrocytes; Direct conversion; Lineage conversion; Neurons; Regeneration; Transdifferentiation; hiPSC
    DOI:  https://doi.org/10.1007/978-1-0716-4530-7_2
  12. Metab Brain Dis. 2025 May 01. 40(5): 196
    Metabolic dysregulation in amyotrophic lateral sclerosis: insights from 1H NMR-based metabolomics in a tertiary care center in India.
    Priyanka Gautam, Rahul Yadav, Ranjeet Kumar Vishwakarma, Abhishek Pathak, Chandan Singh.
      Amyotrophic Lateral Sclerosis (ALS) is a progressive neurodegenerative disorder characterized by motor neuron loss, leading to severe physical impairment and mortality. Despite available treatments like Riluzole and Edaravone, their limited efficacy highlights the need for improved understanding of ALS pathology. This study has explored metabolic alterations in North Indian ALS patients using 1H Nuclear Magnetic Resonance (NMR)-based metabolomics. A case-control study, involving 45 ALS patients and 30 healthy controls (HCs) was performed. Serum samples were analyzed using 600-MHz NMR spectrometer, revealing significant metabolic differences between ALS and HC groups. Multivariate analyses identified nine dysregulated metabolites-pyruvate, glutamine, histidine, isoleucine, leucine, imidazole, arginine, creatinine, and choline-with ROC analysis showing isoleucine as a promising biomarker (AUC 83%). Pathway enrichment analysis highlighted disruptions in key metabolic pathways, including the Glucose-Alanine Cycle, Urea Cycle, Ammonia Recycling, and the Warburg Effect, suggesting potential links to neuroinflammatory and mitochondrial dysfunction in ALS pathogenesis. This pilot study provides insight into ALS-specific metabolic alterations in Indian cohort and demonstrates the potential of these metabolites as diagnostic biomarkers. Our findings identify potential biomarkers that require validation in larger, multi-centric cohorts to support diagnosis, prognosis, and improved management of ALS.
    Keywords:  Amyotrophic lateral sclerosis; Biomarker; Metabolic pathway; Nuclear magnetic resonance; Serum; Untargeted metabolomics
    DOI:  https://doi.org/10.1007/s11011-025-01616-8
  13. Sci Rep. 2025 Apr 29. 15(1): 14976
    Cross-tissue MiRNA profiling of extracellular vesicles and PBMCs from amyotrophic lateral sclerosis patients.
    Dragoni Francesca, Di Gerlando Rosalinda, Diamanti Luca, Bartolo Rizzo, Bordoni Matteo, Scarian Eveljn, Camilla Viola, Cerchia Giovanni, Zucca Susanna, Pansarasa Orietta, Gagliardi Stella.
      RNA-mediated toxicity, which can be controlled by alteration of gene expression, is considered a key event in Amyotrophic Lateral Sclerosis (ALS). Transcriptomic deregulation of miRNAs expression can spread via "horizontal" RNA transfer through extracellular vesicles (EVs) to act in conjunction with proteins, leading to changes in mRNA, which can provide early signals to indicate forthcoming neuropathological changes in the brain. The aim of this work is to compare expression profiles (obtained by miRNA-seq) from different tissues to highlight commonly expressed and tissue-specific miRNAs. miRNA species from plasma EVs were correlated with miRNA profiles obtained from peripheral blood mononuclear cells (PBMCs). Each tissue from ALS patients was compared to controls, revealing 159 deregulated (DE) miRNAs in Exosomes (EXOs), 247 DE miRNAs in PBMCs and 162 DE miRNAs in Microvesicles (MVs). Next, data were filtered to include only miRNAs expressed in disease samples (not in healthy subjects), to reduce the number of tissue- and ALS- specific miRNAs (EXO n = 22, MV = 11, PBMCs n = 8). We identified specific miRNAs and pathways related to each tissue. Interestingly, in PBMCs we found mainly neuro-linked pathways, such as neurotransmitters, brain and neuron development, while in EXOs, we found miRNAs implicated in MAPK and ERB signaling. In contrast, the altered pathways in MVs were not specific. This study shows that the composition of small RNA differs significantly between blood cells and its respective EVs fraction. Differentially expressed miRNAs can target definite transcripts in different cellular and molecular fractions. It is evident that, in terms of miRNAs cargo, MVs are not specific to ALS. Therefore, future studies will focus on the interaction between cells and EXOs.
    Keywords:  ALS; Biomarkers; Extracellular vesicles; MiRNA; PBMCs
    DOI:  https://doi.org/10.1038/s41598-025-99206-2
  14. Mol Neurodegener. 2025 Apr 26. 20(1): 49
    TDP-43 seeding activity in the olfactory mucosa of patients with amyotrophic lateral sclerosis.
    Maria Vizziello, Ilaria Linda Dellarole, Arianna Ciullini, Riccardo Pascuzzo, Annalisa Lombardo, Floriana Bellandi, Luigi Celauro, Claudia Battipaglia, Emilio Ciusani, Ambra Rizzo, Marcella Catania, Grazia Devigili, Sara Adriana Della Seta, Valentina Margiotta, Monica Consonni, Veronica Faltracco, Pietro Tiraboschi, Nilo Riva, Sara Maria Silvia Portaleone, Gianluigi Zanusso, Giuseppe Legname, Giuseppe Lauria, Eleonora Dalla Bella, Fabio Moda.
       BACKGROUND: In recent years, the seed amplification assay (SAA) has enabled the identification of pathological TDP-43 in the cerebrospinal fluid (CSF) and olfactory mucosa (OM) of patients with genetic forms of frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS). Here, we investigated the seeding activity of TDP-43 in OM samples collected from patients with sporadic ALS.
    METHODS: OM samples were collected from patients with (a) sporadic motor neuron diseases (MND), including spinal ALS (n = 35), bulbar ALS (n = 18), primary lateral sclerosis (n = 10), and facial onset sensory and motor neuronopathy (n = 2); (b) genetic MND, including carriers of C9orf72exp (n = 6), TARDBP (n = 4), SQSTM1 (n = 3), C9orf72exp + SQSTM1 (n = 1), OPTN (n = 1), GLE1 (n = 1), FUS (n = 1) and SOD1 (n = 4) mutations; (c) other neurodegenerative disorders (OND), including Alzheimer's disease (n = 3), dementia with Lewy bodies (n = 8) and multiple system atrophy (n = 6); and (d) control subjects (n = 22). All samples were subjected to SAA analysis for TDP-43 (TDP-43_SAA). Plasmatic levels of TDP-43 and neurofilament-light chain (NfL) were also assessed in a selected number of patients.
    RESULTS: TDP-43_SAA was positive in 29/65 patients with sporadic MND, 9/21 patients with genetic MND, 6/17 OND patients and 3/22 controls. Surprisingly, one presymptomatic individual also tested positive. As expected, OM of genetic non-TDP-43-related MND tested negative. Interestingly, fluorescence values from non-MND samples that tested positive were consistently and significantly lower than those obtained with sporadic and genetic MND. Furthermore, among TDP-43-positive samples, the lag phase observed in MND patients was significantly longer than that in non-MND patients. Plasma TDP-43 levels were significantly higher in sporadic MND patients compared to controls and decreased as the disease progressed. Similarly, plasma NfL levels were higher in both sporadic and genetic MND patients and positively correlated with disease progression rate (ΔFS). No significant correlations were detected between TDP-43_SAA findings and the biological, clinical, or neuropsychological parameters considered.
    CONCLUSIONS: The OM of a subset of patients with sporadic MND can trigger seeding activity for TDP-43, as previously observed in genetic MND. Thus, TDP-43_SAA analysis of OM can improve the clinical characterization of ALS across different phenotypes and enhance our understanding of these diseases. Finally, plasma TDP-43 could serve as a potential biomarker for monitoring disease progression. However, further research is needed to confirm and expand these findings.
    Keywords:  Amyotrophic lateral sclerosis; Neurodegeneration; Olfactory mucosa; Peripheral biomarkers; Seed amplification assay; TDP-43
    DOI:  https://doi.org/10.1186/s13024-025-00833-0
  15. NPJ Parkinsons Dis. 2025 Apr 30. 11(1): 103
    The role of alpha-synuclein in synucleinopathy: Impact on lipid regulation at mitochondria-ER membranes.
    Peter A Barbuti, Cristina Guardia-Laguarta, Taekyung Yun, Zena K Chatila, Xena Flowers, Chantel Wong, Bruno F R Santos, Simone B Larsen, James S Lotti, Nobutaka Hattori, Elizabeth Bradshaw, Ulf Dettmer, Saranna Fanning, Vilas Menon, Hasini Reddy, Andrew F Teich, Rejko Krüger, Estela Area-Gomez, Serge Przedborski.
      The protein alpha-synuclein (αSyn) plays a pivotal role in the pathogenesis of synucleinopathies, including Parkinson's disease and multiple system atrophy, with growing evidence indicating that lipid dyshomeostasis is a key phenotype in these neurodegenerative disorders. Previously, we identified that αSyn localizes, at least in part, to mitochondria-associated endoplasmic reticulum membranes (MAMs), which are transient functional domains containing proteins that regulate lipid metabolism, including the de novo synthesis of phosphatidylserine. In the present study, we analyzed the lipid composition of postmortem human samples, focusing on the substantia nigra pars compacta of Parkinson's disease and controls, as well as three less affected brain regions of Parkinson's donors. To further assess synucleinopathy-related lipidome alterations, similar analyses were performed on the striatum of multiple system atrophy cases. Our data reveal region- and disease-specific changes in the levels of lipid species. Specifically, our data revealed alterations in the levels of specific phosphatidylserine species in brain areas most affected in Parkinson's disease. Some of these alterations, albeit to a lesser degree, are also observed in multiple system atrophy. Using induced pluripotent stem cell-derived neurons, we show that αSyn regulates phosphatidylserine metabolism at MAM domains, and that αSyn dosage parallels the perturbation in phosphatidylserine levels. These findings support the notion that αSyn pathophysiology is linked to the dysregulation of lipid homeostasis, which may contribute to the vulnerability of specific brain regions in synucleinopathy. These findings have significant therapeutic implications.
    DOI:  https://doi.org/10.1038/s41531-025-00960-x
  16. Acta Neuropathol. 2025 May 02. 149(1): 42
    Widespread distribution of α-synuclein oligomers in LRRK2-related Parkinson's disease.
    Hiroaki Sekiya, Lukas Franke, Yuki Hashimoto, Mariko Takata, Katsuya Nishida, Naonobu Futamura, Kazuko Hasegawa, Hisatomo Kowa, Owen A Ross, Pamela J McLean, Tatsushi Toda, Zbigniew K Wszolek, Dennis W Dickson.
      Mutations in leucine-rich repeat kinase 2 (LRRK2) are the most common cause of familial and sporadic Parkinson's disease (PD). While the clinical features of patients with LRRK2-PD resemble those of typical PD, there are significant differences in the pathological findings. The pathological hallmark of definite PD is the presence of α-synuclein (αSYN)-positive Lewy-related pathology; however, approximately half of patients with LRRK2-PD do not have Lewy-related pathology. Lewy-related pathology is a late-stage αSYN aggregation that can be visualized with hematoxylin and eosin stains or conventional immunohistochemistry (IHC). Increasing evidence has indicated that αSYN oligomers, which represent the early-stage of αSYN aggregation, may have neurotoxicity. Visualization of αSYN oligomers requires specialized staining techniques, such as αSYN-proximity ligation assay (PLA). Distribution and severity of αSYN oligomers in the brain of patients with LRRK2-PD remain unknown. In this study, we performed phosphorylated αSYN-IHC and αSYN-PLA staining on postmortem brain sections of patients with three pathogenic LRRK2 mutants: p.G2019S (n = 5), p.I2020T (n = 5), and p.R1441C (n = 4). The severity of Lewy-related pathology and αSYN oligomers was assessed semi-quantitatively in the brainstem, limbic lobe, basal ganglia, and cerebral cortex. αSYN oligomers were detected in patients with LRRK2-PD even in those without Lewy-related pathology; a negative correlation was observed between Lewy-related pathology and αSYN oligomers (r = - 0.26 [- 0.39, - 0.12]; P < 0.0001). Our findings suggest that αSYN oligomers may represent a common pathological feature of LRRK2-PD. Notably, patients harboring p.G2019S and p.I2020T had significantly higher levels of αSYN oligomers in those without Lewy-related pathology compared to those with Lewy-related pathology. These patients also had a trend toward shorter disease duration. These results imply that in LRRK2-PD, αSYN oligomers may initially accumulate in the brain but do not progress to form Lewy-related pathology. The present study suggests that targeting αSYN oligomers may be a therapeutic strategy for LRRK2-PD even if there is no Lewy-related pathology.
    Keywords:  Alpha-synuclein; LRRK2; Lewy bodies; Oligomers; Parkinson disease; Pathogenesis
    DOI:  https://doi.org/10.1007/s00401-025-02872-9
  17. bioRxiv. 2025 Apr 10. pii: 2025.04.07.647611. [Epub ahead of print]
    Novel extracellular vesicle release pathway facilitated by toxic superoxide dismutase 1 oligomers.
    Brianna Hnath, Nikolay V Dokholyan.
      Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease resulting in paralysis and death within three to five years. Mutations in over forty different proteins have been linked to ALS, leading to controversy whether ALS is one disease or many diseases with a similar phenotype. Mutations in Cu,Zn superoxide dismutase 1 (SOD1) are only found in 2-3% of ALS cases, yet misfolded SOD1 is found in both sporadic (sALS) and familial (fALS) patients. Yet, mutations in TDP-43 or FUS increase the level of misfolded SOD1 on extracellular vesicles (EVs). Additionally, small EVs isolated from ALS patient samples caused cell death of wild type motor neurons and myotubules. The toxicity and protein alterations of ALS EVs have led to the theory that EVs are responsible for the spread of ALS. We hypothesize that previously-identified toxic trimeric SOD1 is spreading on EVs in ALS and altering the spread of other ALS-related proteins, linking them to a common mechanism. To test our hypothesis, we isolate EVs from motor neuron-like cells expressing trimer stabilizing mutations and perform a sandwich enzyme-linked immunoassay (ELISA) (CD9 capture antibody) to quantify whether misfolded SOD1 and 17 other ALS-related proteins increase or decrease on EVs with trimer stabilization. We identify which EV release pathway is being affected by trimeric SOD1 utilizing endocytosis and exocytosis inhibitors, and determine if any specific EV-related proteins are altered with trimer stabilization. We establish that VAPB, VCP, and Stathmin-2 increase on EVs with trimer stabilization. The common pathway between SOD1 and three other ALS-associated proteins is affected by multiple pathways, including the Caveolae endocytosis pathway, suggesting a novel hybrid pathway of EV release present in ALS.
    DOI:  https://doi.org/10.1101/2025.04.07.647611
  18. Alzheimers Dement. 2025 Apr;21(4): e70198
    Genome-wide association analysis identifies APOE as a mitophagy modifier in Lewy body disease.
    Xu Hou, Michael G Heckman, Fabienne C Fiesel, Shunsuke Koga, Alexandra I Soto-Beasley, Jens O Watzlawik, Jing Zhao, Rebecca R Valentino, Patrick W Johnson, Launia J White, Zachary S Quicksall, Joseph S Reddy, Jose Bras, Rita Guerreiro, Na Zhao, Guojun Bu, Dennis W Dickson, Owen A Ross, Wolfdieter Springer.
       INTRODUCTION: Phosphorylated ubiquitin (p-S65-Ub) is generated during PINK1-PRKN mitophagy as a specific marker of mitochondrial damage. Despite the widespread deposition of p-S65-Ub in aged and diseased human brain, the genetic contribution to its accumulation remains unclear.
    METHODS: To identify novel mitophagy regulators, we performed a genome-wide association study using p-S65-Ub level as a quantitative trait in 1012 autopsy-confirmed Lewy body disease (LBD) samples.
    RESULTS: We identified a significant genome-wide association with p-S65-Ub for rs429358 (apolipoprotein E ε4 [APOE4]) and a suggestive association for rs6480922 (ZMIZ1). APOE4 was associated with higher p-S65-Ub levels and greater neuropathological burden. Functional validation in mouse and human induced pluripotent stem cell (iPSC) models confirmed APOE4-mediated mitophagy alterations. Intriguingly, ZMIZ1 rs6480922 was associated with lower p-S65-Ub levels, reduced neuropathological load, and increased brain weight, indicating a potential protective role.
    DISCUSSION: Our findings underscore the importance of mitochondrial quality control in LBD pathogenesis and nominate regulators that may contribute to disease risk or resilience.
    HIGHLIGHTS: p-S65-Ub levels were used as a quantitative marker of mitochondrial damage. A GWAS identified two genetic variants that modify mitophagy in LBD autopsy brain. APOE4 was associated with increased p-S65-Ub accumulation and neuropathology. APOE4 altered mitophagy via pathology-dependent and pathology-independent mechanisms. ZMIZ1 was linked to reduced p-S65-Ub and neuropathology indicative of protection.
    Keywords:  GWAS; PINK1; PRKN; Parkin; Parkinson's disease; ZMIZ1; autophagy; mitochondria; phosphorylated ubiquitin; ubiquitin
    DOI:  https://doi.org/10.1002/alz.70198
  19. bioRxiv. 2025 Apr 09. pii: 2024.04.01.587651. [Epub ahead of print]
    Nanoscale domains govern local diffusion and aging within FUS condensates.
    Guoming Gao, Emily R Sumrall, Nils G Walter.
      Biomolecular condensates regulate cellular physiology by sequestering and processing RNAs and proteins, yet how these processes are locally tuned within condensates remains unclear. Moreover, in neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS), condensates undergo liquid-to-solid phase transitions, but capturing early intermediates in this process has been challenging. Here, we present a surface multi-tethering approach to achieve intra-condensate single-molecule tracking of fluorescently labeled RNA and protein molecules within liquid-like condensates. Using RNA-binding protein Fused in Sarcoma (FUS) as a model for condensates implicated in ALS, we discover that RNA and protein diffusion is confined within distinct nanometer-scale domains, or nanodomains, which exhibit unique connectivity and chemical environments. During condensate aging, these nanodomains reposition, facilitating FUS fibrilization at the condensate surface, a transition enhanced by FDA-approved ALS drugs. Our findings demonstrate that nanodomain formation governs condensate function by modulating biomolecule sequestration and percolation, offering insights into condensate aging and disease-related transitions.
    DOI:  https://doi.org/10.1101/2024.04.01.587651
  20. Contact (Thousand Oaks). 2025 Jan-Dec;8:8 25152564251321770
    Profiling the LAM Family of Contact Site Tethers Provides Insights into Their Regulation and Function.
    Emma J Fenech, Meital Kupervaser, Angela Boshnakovska, Shani Ravid, Inês Gomes Castro, Yeynit Asraf, Sylvie Callegari, Christof Lenz, Henning Urlaub, Peter Rehling, Maya Schuldiner.
      Membrane contact sites are molecular bridges between organelles that are sustained by tethering proteins and enable organelle communication. The endoplasmic reticulum (ER) membrane harbors many distinct families of tether proteins that enable the formation of contacts with all other organelles. One such example is the LAM (Lipid transfer protein Anchored at Membrane contact sites) family in yeast, which is composed of six members, each containing a putative lipid binding and transfer domain and an ER-embedded transmembrane segment. The family is divided into three homologous pairs each unique in their molecular architecture and localization to different ER subdomains. However, what determines the distinct localization of the different LAMs and which specific roles they carry out in each contact are still open questions. To address these, we utilized a labeling approach to profile the proximal protein landscape of the entire family. Focusing on unique, candidate interactors we could support that Lam5 resides at the ER-mitochondria contact site and demonstrate a role for it in sustaining mitochondrial activity. Capturing shared, putative interactors of multiple LAMs, we show how the Lam1/3 and Lam2/4 paralogous pairs could be associated specifically with the plasma membrane. Overall, our work provides new insights into the regulation and function of the LAM family members. More globally it demonstrates how proximity labeling can help identify the shared or unique functions of paralogous proteins.
    Keywords:  ABOLISH; GRAMD/ASTER/STARD; LAM protein family; endoplasmic reticulum; membrane contact sites; proximity labeling
    DOI:  https://doi.org/10.1177/25152564251321770
  21. Cell Mol Life Sci. 2025 Apr 28. 82(1): 188
    The potential of brain organoids in addressing the heterogeneity of synucleinopathies.
    Xiao-Jun Diao, Claudio Soto, Fei Wang, Yu Wang, Yun-Cheng Wu, Abhisek Mukherjee.
      Synucleinopathies are a group of diseases characterized by neuronal and glial accumulation of α-synuclein (aSyn) linked with different clinical presentations, including Parkinson's disease (PD), Parkinson's disease with dementia (PDD), Dementia with Lewy Bodies (DLB) and Multiple system atrophy (MSA). Interestingly, the structure of the aSyn aggregates can vary across different synucleinopathies. Currently, it is unclear how the aSyn protein can aggregate into diverse structures and affect distinct cell types and various brain regions, leading to different clinical symptoms. Recent advances in induced pluripotent stem cells (iPSCs)-based brain organoids (BOs) technology provide an unprecedented opportunity to define the etiology of synucleinopathies in human brain cells within their three-dimensional (3D) context. In this review, we will summarize current advances in investigating the mechanisms of synucleinopathies using BOs and discuss the scope of this platform to define mechanisms underlining the selective vulnerability of cell types and brain regions in synucleinopathies.
    Keywords:  Cerebral organoids; Midbrain organoids; Parkinson’s disease; Pluripotent stem cells; Synucleinopathies
    DOI:  https://doi.org/10.1007/s00018-025-05686-w
  22. Mol Med. 2025 Apr 29. 31(1): 158
    Preclinical evaluation of AAV9-coSMN1 gene therapy for spinal muscular atrophy: efficacy and safety in mouse models and non-human primates.
    Wenhao Ma, Zhijie Wu, Tianyi Zhao, Yan Xia, Jing Qin, Xue Tian, Xin Li, Jun He, Yan Zhang, Lina Zhang, Li Li, Zheyue Dong, Zhichun Feng, Xiaoyan Dong, Wang Sheng, Xiaobing Wu.
       BACKGROUND: Spinal muscular atrophy (SMA) is a severe neuromuscular disorder caused by the loss of motor neurons in the spinal cord. Our team has initiated clinical trials using adeno-associated virus serotype 9 (AAV9) vectors carrying a codon-optimized human SMN1 (coSMN1) gene, delivered via intrathecal (IT) injection. Here, we present the preclinical research that laid the groundwork for these trials, offering comprehensive data on the efficacy and safety of AAV9-coSMN1 in both murine models and non-human primates.
    MATERIAL AND METHOD: We developed a codon-optimized hSMN1 expression cassette and analyzed SMN protein levels using Western blot and immunofluorescence. Taiwanese SMA-like mouse model was employed to assess tail length preservation, as well as to examine motor neuron and skeletal muscle pathological phenotypes through immunofluorescence and histopathological staining. Serum biomarkers in both mice and cynomolgus monkeys were measured using a blood chemistry analyzer. The in-vivo biodistribution of AAV9-coSMN1 and toxicological profile were investigated through quantitative Polymerase Chain Reaction(qPCR) and histopathological staining.
    RESULTS: Codon optimization of hSMN1 led to enhanced gene expression and increased SMN protein levels in vitro. AAV9-coSMN1 demonstrated significant therapeutic efficacy in a Type 3 SMA mouse model, effectively rescuing motor neurons, preserving tail integrity, and improving skeletal muscle histopathology. In vivo studies, both mice and cynomolgus monkeys revealed widespread CNS distribution following a single intracerebroventricular or intrathecal injection, with no observed toxic inflammatory responses in the dorsal root ganglia. Peripheral organs also showed detectable levels of the vector gene, indicating effective systemic distribution.
    CONCLUSION: The preclinical evaluation confirms that AAV9-coSMN1 is a safe and effective therapeutic candidate for SMA, with potential applicability across various phenotypes. The study provides critical data supporting its advancement to clinical trials, underscoring its promise for broader neurological applications.
    DOI:  https://doi.org/10.1186/s10020-025-01207-4
  23. Autophagy Rep. 2025 ;pii: 2484835. [Epub ahead of print]4(1):
    Image-based temporal profiling of autophagy-related phenotypes.
    Nitin Sai Beesabathuni, Neil Alvin B Adia, Eshan Thilakaratne, Ritika Gangaraju, Priya S Shah.
      Autophagy is a dynamic process critical in maintaining cellular homoeostasis. Dysregulation of autophagy is linked to many diseases and is emerging as a promising therapeutic target. High-throughput methods to characterise autophagy are essential for accelerating drug discovery and characterising mechanisms of action. In this study, we developed a scalable image-based temporal profiling approach to characterise ~900 morphological features at a single cell level with high temporal resolution. We differentiated drug treatments based on morphological profiles using a random forest classifier with ~90% accuracy and identified the key features that govern classification. Additionally, temporal morphological profiles accurately predicted biologically relevant changes in autophagy after perturbation, such as total cargo degraded. Therefore, this study acts as proof-of-principle for using image-based temporal profiling to differentiate autophagy perturbations in a high-throughput manner and has the potential identify biologically relevant autophagy phenotypes. Ultimately, approaches like image-based temporal profiling can accelerate drug discovery.
    Keywords:  Autophagy; autophagy flux; cargo degradation; fluorescence microscopy; live cell imaging; morphological features; rapamycin; temporal profiling; wortmannin
    DOI:  https://doi.org/10.1080/27694127.2025.2484835
  24. Sci Rep. 2025 Apr 30. 15(1): 15205
    Glutamate gradually elevates [Zn2+]i via the CaM-CaMKII-NOS cascade in primary cultured rat embryonic cortical neurons.
    Hui-Chiun Tseng, Yong-Sheng Wang, Chien-Yuan Pan.
      Zn2+ is essential for neuronal signaling, but imbalance cause cell death and neurodegenerative disorders. While the buffering system maintains low cytosolic Zn2+ concentration ([Zn2+]i), the details on physiological stimuli elevating [Zn2+]i for neuronal processes remain limited. Our previous reports have demonstrated that dopamine elevates [Zn2+]i through the cAMP-NO pathway, activating autophagy and inflammation in neurons. In this study, we adopted the Zn2+ imaging technique to verify how glutamate elevated [Zn2+]i in cultured cortical neurons and examined the inflammatory response. Our results showed that glutamate elevates the [Zn2+]i, by activating ionotropic glutamate receptors. Inhibitors of calmodulin (CaM), CaM-dependent protein kinase II (CaMKII), and NO synthase (NOS) blocked the glutamate-induced Zn2+ response. High-K+ buffer induced-membrane depolarization significantly elevated the intracellular Ca2+ concentration ([Ca2+]i) but only slightly increased [Zn2+]i and NO production. Glutamate also transiently increased NOS phosphorylation at Ser1417 within 15 min. The Zn2+ chelator, TPEN suppressed glutamate-induced inflammasome formation. These results indicate that glutamate-induced local increment in [Ca2+]i via the ionotropic glutamate receptors activates the CaM-CaMKII-NOS complex to produce NO and elevate [Zn2+]i. which trigger inflammation in cultured neurons. Henceforth, this novel glutamate-Zn2+ signaling pathway after glutamate depolarization elevates [Ca2+]i indicates the involvement of Zn2+ in modulating long-term neuronal activities.
    Keywords:  Calmodulin; Calmodulin-dependent protein kinase II; Inflammation; Ionotropic glutamate receptor; Nitric oxide synthase; Zn2+
    DOI:  https://doi.org/10.1038/s41598-025-99142-1
  25. Antioxidants (Basel). 2025 Mar 28. pii: 401. [Epub ahead of print]14(4):
    Dysfunctional Mitochondria Characterize Amyotrophic Lateral Sclerosis Patients' Cells Carrying the p.G376D TARDBP Pathogenetic Substitution.
    Giuseppe Petito, Victoria Stefania Del Fiore, Arianna Cuomo, Federica Cioffi, Gilda Cobellis, Antonia Lanni, Flora Guerra, Cecilia Bucci, Rosalba Senese, Roberta Romano.
      Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease caused by the degeneration of upper and lower motor neurons in the brain, brainstem and spinal cord. About 10% of familial ALS cases are linked to pathogenetic substitution in TARDBP, the gene encoding the TDP-43 protein. A novel rare causative variant in TARDBP (p.G376D) was recently reported in ALS patients. It leads to TDP-43 cytoplasmic mislocalization, increased oxidative stress and reduced cell viability. However, functional studies on the effects of this molecular defect have not yet been carried out. Mitochondria are highly dynamic organelles, and their deregulation has emerged as a key factor in many diseases, among which is ALS. Therefore, this study aimed at determining the impact of this causative variant on mitochondria. In cellular models expressing TDP-43G376D and in fibroblasts derived from patients carrying this molecular defect, we observed alterations of mitochondrial functionality. We demonstrated increased localization of the mutated protein to mitochondria and a reduced abundance of subunits of complex I and complex II of the mitochondrial respiratory chain, associated with a decrease in mitochondrial membrane potential, in cellular respiration and in cytochrome C oxidase (COX) activity. Moreover, ALS cells showed increased mitochondrial fragmentation and reduced abundance of antioxidant enzymes causing increased oxidative stress. These results expand our knowledge about the molecular mechanisms underlying ALS pathogenesis associated with TDP-43 p.G376D and could help to identify new therapeutic strategies to counteract this disease.
    Keywords:  TARDBP; TDP-43; amyotrophic lateral sclerosis; mitochondria; oxidative stress
    DOI:  https://doi.org/10.3390/antiox14040401
  26. Biomolecules. 2025 Mar 24. pii: 473. [Epub ahead of print]15(4):
    Immunological Fluid Biomarkers in Frontotemporal Dementia: A Systematic Review.
    Philip Ahle Erichsen, Emil Elbæk Henriksen, Jørgen Erik Nielsen, Patrick Ejlerskov, Anja Hviid Simonsen, Anders Toft.
      Dysregulated immune activation plays a key role in the pathogenesis of neurodegenerative diseases, including frontotemporal dementia (FTD). This study reviews immunological biomarkers associated with FTD and its subtypes. A systematic search of PubMed and Web of Science was conducted for studies published before 1 January 2025, focusing on immunological biomarkers in CSF or blood from FTD patients with comparisons to healthy or neurological controls. A total of 124 studies were included, involving 6686 FTD patients and 202 immune biomarkers. Key findings include elevated levels of GFAP and MCP1/CCL2 in both CSF and blood and consistently increased CHIT1 and YKL-40 in CSF. Complement proteins from the classical activation pathway emerged as promising targets. Distinct immune markers were found to differentiate FTD from Alzheimer's disease (AD) and amyotrophic lateral sclerosis (ALS), with GFAP, SPARC, and SPP1 varying between FTD and AD and IL-15, HERV-K, NOD2, and CHIT1 differing between FTD and ALS. A few markers, such as Galectin-3 and PGRN, distinguished FTD subtypes. Enrichment analysis highlighted IL-10 signaling and immune cell chemotaxis as potential pathways for further exploration. This study provides an overview of immunological biomarkers in FTD, emphasizing those most relevant for future research on immune dysregulation in FTD pathogenesis.
    Keywords:  Alzheimer’s disease; amyotrophic lateral sclerosis; biomarkers; frontotemporal dementia; immune
    DOI:  https://doi.org/10.3390/biom15040473
  27. ACS Chem Neurosci. 2025 May 01.
    Deciphering the Inhibitory Mechanism of ALS-Associated N352S and S352p Variants against TDP-43 Aggregation and Its Destabilization Effect on TDP-43 Protofibrils.
    Zhengdong Xu, Wenjuan Yi, Lulu Guan, Jiaxing Tang, Dushuo Feng, Yu Zou.
      Amyotrophic lateral sclerosis (ALS) is closely related to ubiquitin-positive inclusions formed by transactive response deoxyribonucleic acid (DNA) binding protein of 43 kDa (TDP-43). Previous experiments identified that the ALS-linked familial variant, N352S (asparagine substituted by serine), and subsequent phosphorylation of S352 (S352p) are associated with the aggregation of TDP-43. However, the underlying molecular mechanisms are still not fully understood. By performing all-atom explicit-solvent replica exchange molecular dynamics (REMD) simulations with a total simulation time of 100.8 μs, we scrutinized the impact of the N352S mutation and its phosphorylation variant S352p on the conformational ensembles of the TDP-43342-366 dimer. Our simulation results show that both the N352S and S352p variants could promote the formation of unstructured conformation and impede the formation of β-structure and helix content, and the inhibitive effect of S352P is more obvious. Further analyses suggest that the H-bonding and hydrophobic interaction among TDP-43342-366 peptides, as well as the R361-E362 salt bridge, are attenuated by N352S and S352p variants. Additional MD simulations show that N352S and S352p variants reduce the structural stability of the hydrophobic region and lower the number of H-bonds and contacts of two hydrophobic clusters, thus possessing a destabilization effect on the TDP-43282-360 protofibrils. Our results unmask the molecular mechanism of the N352S mutation and its phosphorylation variant S352p toward the inhibition of TDP-43342-366 aggregation and prove the protofibril-destabilizing effects of these two variants, which may be helpful for designing drugs for the treatment of ALS.
    Keywords:  N352S mutation; TDP-43; aggregation; amyotrophic lateral sclerosis; molecular dynamics simulation; phosphorylation
    DOI:  https://doi.org/10.1021/acschemneuro.5c00045
  28. Contact (Thousand Oaks). 2025 Jan-Dec;8:8 25152564251332141
    Mitochondria-Plasma Membrane Contact Sites: Emerging Regulators of Mitochondrial Form and Function.
    Jason C Casler, Matilde V Neto, Thomas Burgoyne, Laura L Lackner.
      Sites of close apposition between organelles, known as membrane contact sites (MCSs), are critical regulators of organelle function. Mitochondria form elaborate reticular networks that perform essential metabolic and signaling functions. Many mitochondrial functions are regulated by MCSs formed between mitochondria and other organelles. In this review, we aim to bring attention to an understudied, but physiologically important, MCS between mitochondria and the plasma membrane (PM). We first describe the molecular mechanism of mitochondria-PM tethering in budding yeast and discuss its role in regulating multiple biological processes, including mitochondrial dynamics and lipid metabolism. Next, we discuss the evidence for mitochondria-PM tethering in higher eukaryotes, with a specific emphasis on mitochondria-PM contacts in retinal cells, and speculate on their functions. Finally, we discuss unanswered questions to guide future research into the function of mitochondria-PM contact sites.
    Keywords:  cell biology; electron microscopy; interorganelle (inter-organelle); membrane contact sites (MCSs)‌; mitochondrion (mitochondria); plasma membrane
    DOI:  https://doi.org/10.1177/25152564251332141
  29. Biomedicines. 2025 Apr 13. pii: 952. [Epub ahead of print]13(4):
    The Role of mTOR in Amyotrophic Lateral Sclerosis.
    José Augusto Nogueira-Machado, Fabiana Rocha-Silva, Nathalia Augusta Gomes.
      Background: Amyotrophic lateral sclerosis (ALS) is a rare, progressive, and incurable disease characterized by muscle weakness and paralysis. Recent studies have explored a possible link between ALS pathophysiology and mTOR signaling. Recent reports have linked the accumulation of protein aggregates, dysfunctional mitochondria, and homeostasis to the development of ALS. mTOR plays a pivotal role in controlling autophagy and affecting energy metabolism, in addition to supporting neuronal growth, plasticity, and the balance between apoptosis and autophagy, all of which are important for homeostasis. Aim: This mini-review approaches the regulatory roles of mTOR signaling pathways, their interaction with other metabolic pathways, and their potential to modulate ALS progression. Significance: It discusses how these metabolic signaling pathways affect the neuromuscular junction, producing symptoms of muscle weakness and atrophy similar to those seen in patients with ALS. The discussion includes the concepts of neurocentric and peripheral and the possible connection between mTOR and neuromuscular dysfunction in ALS. Conclusions: It highlights the therapeutic potential of mTOR signaling and interconnections with other metabolic routes, making it a promising biomarker and therapeutic target for ALS.
    Keywords:  ALS; amyotrophic lateral sclerosis; genetic biomarkers; mTOR; metabolic signaling; neurodegenerative diseases; neuromuscular degeneration
    DOI:  https://doi.org/10.3390/biomedicines13040952
  30. Biomolecules. 2025 Apr 15. pii: 585. [Epub ahead of print]15(4):
    A Novel Phosphorylated Tau Conformer Implicated in the Tauopathy Pathogenesis of Human Neurons.
    Nahid Tofigh, Sadaf Agahi, Gholamhossein Riazi, Mahboobeh Ghalamkar Moazzam, Koorosh Shahpasand.
      Alzheimer's disease (AD) is a neurodegenerative disorder with no effective treatments. Hyperphosphorylation of tau protein contributes to neurodegeneration in AD. Previous studies have identified pT231-tau in the cis conformation as an early driver of neurodegeneration in tauopathy models. Here, we identify a novel neurotoxic pT231-tau conformer in human AD neurons, distinct from both cis and trans conformations, which we propose as the gauche pT231-tau conformer. Notably, levels of this conformer were elevated in neurons subjected to aging-associated stress. In order to confirm the stress, we examined p21 accumulation in both human iPSC-derived and mouse cortical neurons under aging stress. Targeted elimination of the gauche pT231-tau conformer mitigated neurodegeneration in human AD cultures. These findings suggest the gauche pT231-tau conformer plays a key role in tau-mediated neurodegeneration and may be a potential therapeutic target for AD.
    Keywords:  Alzheimer’s disease; gauche p-tau; neurodegeneration; pT231-tau monoclonal antibody
    DOI:  https://doi.org/10.3390/biom15040585
  31. Mamm Genome. 2025 Apr 29.
    Challenges of modelling TDP-43 pathology in mice.
    José Miguel Brito Armas, Lucas Taoro-González, Elizabeth M C Fisher, Abraham Acevedo-Arozena.
      TDP-43 is a normally nuclear RNA binding protein that under pathological conditions may be excluded from the nucleus and deposited in the cytoplasm in the form of insoluble polyubiquitinated and polyphosphorylated inclusions. This nuclear exclusion coupled with cytoplasmic accumulation is called TDP-43 pathology and contributes to a range of disorders collectively known as TDP-43 proteinopathies. These include the great majority of amyotrophic lateral sclerosis (ALS) cases, all limbic-predominant age-related TDP-43 encephalopathy (LATE), as well as up to 50% of frontotemporal lobar degeneration (FTLD) and Alzheimer's disease (AD) cases. Thus, TDP-43 pathology is a common feature underlying a wide range of neurodegenerative conditions. However, modelling it has proven to be challenging, particularly generating models with concomitant TDP-43 loss of nuclear function and cytoplasmic inclusions. Here, focussing exclusively on mice, we discuss TDP-43 genetic models in terms of the presence of TDP-43 pathology, and we consider other models with TDP-43 pathology due to mutations in disparate genes. We also consider manipulations aimed at producing TDP-43 pathology, and we look at potential strategies to develop new, much needed models to address the many outstanding questions regarding how and why TDP-43 protein leaves the nucleus and accumulates in the cytoplasm, causing downstream dysfunction and devastating disease.
    DOI:  https://doi.org/10.1007/s00335-025-10131-1
  32. Arch Toxicol. 2025 Apr 28.
    Application of a high-density microelectrode array assay using a 3D human iPSC-derived brain microphysiological system model for in vitro neurotoxicity screening of environmental compounds.
    Kelly E Carstens, Elena Gronskaya, David Jäckel, Jessica Bertoli, Kelvin Ramirez Cuevas, Julien Dorier, Shan Wang, David Lopez-Rodriguez, Timothy J Shafer, Marie-Gabrielle Zurich, David Pamies.
      Unraveling the associations between human exposure to environmental chemicals and potential neurotoxicity presents significant challenges. Evaluation of neurotoxicity potential using animal testing is resource-intensive (financial, labor, and animal use) and faces uncertainties regarding biological relevance to human health outcomes. Therefore, there is a need to develop efficient and human-relevant in vitro new approach methodologies (NAMs) to screen and evaluate chemicals for neurotoxicity potential. Recording of neural network activity using microelectrode array (MEA) technology has been identified as a reliable and reproducible method for evaluating neurotoxicity. Much of this research has been performed in 2D rodent-derived cell models. The 'BrainSpheres MEA assay' described in this study offers a promising functional human induced pluripotent stem cell (iPSC)-derived 3D brain model comprising neurons, astrocytes, and oligodendrocytes. We demonstrate consistent spontaneous neuronal firing and network bursting parameters from 7-week-old BrainSpheres using a high-density MEA technology. The performance of this model as a human-relevant NAM was evaluated by conducting a multi-concentration, 13 day exposure study with a set of ten chemicals. Neural activity metrics were assessed and compared to results from a 2D-MEA assay using rodent cells. Loperamide and domoic acid (two assay positive controls) demonstrated similar bioactivity profiles in the BrainSphere MEA assay to the 2D-MEA assay, while acetaminophen (assay negative control) was inactive in both assays. The 2D-MEA model demonstrated more potent bioactivity for 4/7 chemicals that were active in both assays. In the future, reducing replicate variability and testing a larger set of chemicals will likely improve the accuracy and reliability of the assay. These preliminary findings suggest that the BrainSphere assay could be used alongside the rat network formation assay (rNFA) as part of a tiered strategy, where hits in the rNFA are confirmed and further characterized in the BrainSphere model, helping move toward animal-free toxicological testing.
    Keywords:  3D cultures; BrainSphere; High-density electrode array; MEA; MPS; Microelectrode array; Neuronal electrical activity; Neurotoxicology; iPSC
    DOI:  https://doi.org/10.1007/s00204-025-04043-x
  33. Front Cell Dev Biol. 2025 ;13 1516596
    Differences in neuronal ciliation rate and ciliary content revealed by systematic imaging-based analysis of hiPSC-derived models across protocols.
    Walther Haenseler, Melanie Eschment, Beth Evans, Marta Brasili, Joana Figueiro-Silva, Fee Roethlisberger, Affef Abidi, Darcie Jackson, Martin Müller, Sally A Cowley, Ruxandra Bachmann-Gagescu.
       Introduction: Ciliopathies are a group of human Mendelian disorders caused by dysfunction of primary cilia, small quasi-ubiquitous sensory organelles. Patients suffering from ciliopathies often display prominent neurodevelopmental phenotypes, underscoring the importance of primary cilia during development and for function of the central nervous system (CNS). Human tissues, in particular from the CNS, are very hard to obtain for research. Patient derived- or genetically engineered human induced pluripotent stem cells (hiPSCs) are therefore a precious resource for investigating the role of cilia in human neurons.
    Methods: In this study we used a variety of 2D and 3D neuronal differentiation protocols in multiple hiPSC lines and systematically analyzed ciliation rates and ciliary length in hiPSCs, neural stem cells (NSCs), immature and different types of mature neurons using immunofluorescence.
    Results: We found that ciliation rate varied substantially between cell lines and differentiation protocols. Moreover, ciliation rate depended on differentiation stage, being maximal in NSCs and decreasing with neuronal maturation. In various types of mature neurons obtained with different protocols, we found ciliation rates to be as low as ∼10%. Neuronal density also played an important role, with higher ciliation in denser cultures. We further investigated the ciliary protein content in these cells at different differentiation stages using commonly used antibodies against ARL13B, INPP5E, AC3 and GPR161. Cilia in hiPSCs, NSCs and neurons were all positive for ARL13B, with a decreasing trend in intensity in more mature neurons. Likewise, INPP5E was present in all cilia analyzed, while AC3 positivity increased as maturation proceeded. Interestingly, we found that while GPR161 signal almost completely disappeared from cilia upon Sonic hedgehog (SHH) stimulation in NSCs and immature neurons, this was not the case in more mature neurons, suggesting a possible developmental time window for cilia-dependent SHH signaling.
    Conclusion: Taken together, our results provide a systematic description of cilia in hiPSC-derived neuronal cells generated with different protocols, underscoring the importance of selecting the optimal model system and controls for investigating primary cilia in hiPSC-derived neuronal cells.
    Keywords:  cilia; ciliopathies; human iPSC (induced pluripotent stem cells); immunofluorescence staining; neurons
    DOI:  https://doi.org/10.3389/fcell.2025.1516596
  34. Pharmaceuticals (Basel). 2025 Apr 03. pii: 524. [Epub ahead of print]18(4):
    Elucidating the Synergistic Effect of the PrimeC Combination for Amyotrophic Lateral Sclerosis in Human Induced Pluripotent Stem Cell-Derived Motor Neurons and Mouse Models.
    Shiran Salomon-Zimri, Nitai Kerem, Gabriel R Linares, Niva Russek-Blum, Justin K Ichida, Ferenc Tracik.
      Background: Amyotrophic lateral sclerosis (ALS) is a multifactorial neurodegenerative disease characterized by the involvement of multiple pathways and mechanisms. The complexity of its pathophysiology is reflected in the diverse hypotheses relating to its underlying causes. Given this intricate interplay of processes, a combination therapy approach offers a promising strategy. Combination therapies have demonstrated significant success in treating complex diseases, where they aim to achieve synergistic therapeutic effects and reduce drug dosage. PrimeC is an oral combination treatment composed of a patented novel formulation consisting of specific and unique doses of two well-characterized drugs (ciprofloxacin and celecoxib). It aims to synergistically inhibit the progression of ALS by addressing key elements of its pathophysiology. Objectives: Demonstrating the synergistic effect of the PrimeC combination compared to each of its individual components, celecoxib and ciprofloxacin, and assessing its ability to improve the drug concentration profile and efficacy. Methods: The efficacy of the PrimeC combination was assessed in a survival assay using human induced pluripotent stem cell (iPSC)-derived motor neurons. Additionally, a drug profiling study was conducted, measuring drug levels in the brain and serum of C57BL mice treated with a single compound versus the combination. Results: Motor neurons modeling ALS treated with the PrimeC combination exhibited better survival rates compared to treatment with either individual compound alone. The enhanced efficacy of the combination was further supported by a drug concentration profiling study in rodents, demonstrating that the PrimeC combination resulted in increased ciprofloxacin concentrations in both brain tissue and serum-highlighting the optimized interaction and synergistic potential of its two comprising agents. Conclusions: Our findings support the potential of combination therapy as an effective strategy for ALS treatment. Specifically, the PrimeC combination demonstrated promising therapeutic effects, providing a strong rationale for its ongoing development as a targeted treatment for ALS.
    Keywords:  amyotrophic lateral sclerosis; combination therapy; iPSCs; in vitro; in vivo; mechanism of action; preclinical; synergism
    DOI:  https://doi.org/10.3390/ph18040524
  35. Brain Sci. 2025 Mar 22. pii: 329. [Epub ahead of print]15(4):
    VGF and Its Derived Peptides in Amyotrophic Lateral Sclerosis.
    Antonio Luigi Manai, Paola Caria, Barbara Noli, Cristina Contini, Barbara Manconi, Federica Etzi, Cristina Cocco.
      Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by a progressive degeneration in the neurons of the frontal cortex, spinal cord, and brainstem, altering the correct release of neurotransmitters. The disease affects every muscle in the body and could cause death three to five years after symptoms first occur. There is currently no efficient treatment to stop the disease's progression. The lack of identification of potential therapeutic strategies is a consequence of the delayed diagnosis due to the absence of accurate ALS early biomarkers. Indeed, neurotransmitters altered in ALS are not measurable in body fluids at quantities that allow for testing, making their use as diagnostic tools a challenge. Contrarily, neuroproteins and neuropeptides are chemical messengers produced and released by neurons, and most of them have the potential to enter bodily fluids. To find out new possible ALS biomarkers, the research of neuropeptides and proteins is intensified using mass spectrometry and biochemical-based assays. Neuropeptides derived from the proVGF precursor protein act as signaling molecules within neurons. ProVGF and its derived peptides are expressed in the nervous and endocrine systems but are also widely distributed in body fluids such as blood, urine, and cerebrospinal fluid, making them viable options as disease biomarkers. To highlight the proVGF and its derived peptides' major roles as ALS diagnostic biomarkers, this review provides an overview of the VGF peptide alterations in spinal cord and body fluids and outlines the limitations of the reported investigations.
    Keywords:  ALS; biomarkers; neuropeptide; proVGF; proteins
    DOI:  https://doi.org/10.3390/brainsci15040329
  36. medRxiv. 2025 Apr 25. pii: 2025.04.23.25326161. [Epub ahead of print]
    Skeletal muscle biomarkers of amyotrophic lateral sclerosis: a large-scale, multi-cohort proteomic study.
    Oleksandr Dergai, Joanne Wuu, Magdalena Koziczak-Holbro, Andrea Malaspina, Volkan Granit, Jessica P Hernandez, Anne Cooley, Ruchika Sachdev, Lili Yu, Michael Bidinosti, Ludvine Flotte, Mark Nash, Lori L Jennings, James D Berry, Lucie I Bruijn, Sophie Brachat, Michael Benatar.
       Background: Biomarkers with clear contexts-of-use are important tools for ALS therapy development. Understanding their longitudinal trajectory in the untreated state is key to their use as potential markers of pharmacodynamic response. To this end, we undertook a large-scale proteomic study in well-phenotyped cohorts to identify biomarker candidates of ALS disease state and disease progression.
    Methods: Clinical phenotypic data and biofluid samples, collected from patients with ALS and healthy controls through multiple longitudinal natural history studies, were used to identify biomarker candidates. SOMAmer (Slow Off-rate Modified Aptamer)-based relatively quantitative measurement of ∼7,000 proteins was performed in plasma and CSF, with immunoassay validation of candidates of interest.
    Results: We identified 329 plasma proteins significantly differentially regulated between ALS and controls (adjusted p-value <0.05), with 25 showing >40% relative abundance. PDLIM3, TNNT2, and MYL11 had the greatest log-fold elevation, while ANTXR2 and ART3 had the greatest log-fold reduction. A similar set of plasma proteins was found to increase (e.g. PDLIM3, TNNT2, MYL11) or decrease (e.g. ANTXR2, ART3, MSTN) with disease progression. CSF proteins with the greatest log-fold elevation included NEFL, NEFH, CHIT1, CA3, MYL11 and GPNMB. These results were confirmed in an independent replication cohort. Moreover, tissue-specific signature enrichment suggests a significant contribution of muscle as a source of these biomarkers. Immunoassays provided orthogonal validation of plasma TNNT2 and CSF GPNMB.
    Conclusion: We identified an array of novel biomarkers with the potential to serve as response biomarkers to aid therapy development, as well as to shed light on the underlying biology of disease.
    Key messages: What is already known on this topic: There are currently few monitoring and disease progression biomarkers in ALS; and there is no published work from large-scale, multi-cohort proteomic studies that utilized longitudinal plasma and CSF samples to help fill this gap.What this study adds: Using Slow Off-rate Modified Aptamer (SOMAmer)-based methods, we have identified an array of novel biomarkers of disease state (i.e. differentially regulated in ALS vs. controls) and ALS disease progression. These included, among others, PDLIM3, MYL11, ANTXR2, ART3, and MSTN.Skeletal muscle is the likely source of many of these newly discovered biomarkers.How this study might affect research, practice or policy: These newly identified monitoring and disease progression biomarkers may be used to evaluate pharmacodynamic response in future clinical trials, thereby aiding ALS therapy development efforts.
    DOI:  https://doi.org/10.1101/2025.04.23.25326161
  37. Brain. 2025 Apr 28. pii: awaf130. [Epub ahead of print]
    Emerging roles of transfer RNA fragments in the CNS.
    Katarzyna Winek, Hermona Soreq.
      tRNA-derived small RNAs (tsRNAs), previously considered inactive tRNA degradation products, have now been shown to be functional small noncoding RNAs. They may play important roles within the central nervous system (CNS) and in brain-body interactions both during normal developmental stages as well as in diverse brain pathologies. Among the cell types found in the CNS, tsRNAs are most abundant in neurons. Correspondingly, neurons show cell type specific tRNA expression profiles when compared to other cells of the CNS under homeostatic conditions and defects in tRNA processing may lead to neurological disorders. Disease-specific tsRNA profiles have been identified in a number of CNS disorders including amyotrophic lateral sclerosis (ALS) and epilepsy. Elevated levels of specific tsRNAs have been found in the blood before the onset of epileptic seizures, and age-related, sex-specific loss of mitochondrial genome-originated tsRNAs in the nucleus accumbens of female patients is correlated to accelerated cognitive deterioration in Alzheimer's disease. Disease-related tsRNA signatures have also been identified in the cerebrospinal fluid of Parkinson's disease patients, and nucleated blood cells from ischemic stroke patients show specific elevation of cholinergic-targeted tsRNAs. The mechanisms of action of tsRNAs are still being elucidated but include targeting complementary mRNA to impact RNA levels and translation in a miRNA-like manner, direct interaction with RNA binding proteins, or interference with translation machinery. The function of tsRNAs may be affected by the chemical modifications they inherit from the originating tRNA molecules, which impact tsRNAs production and may modulate their interactions with proteins. Research on the genetics, biochemical properties and regulatory roles of tsRNAs has expanded rapidly in recent years, facilitated by novel sequencing strategies which include the removal of tRNA modifications and chemically blocked ends that hinder amplification and adapter ligation. Future in-depth profiling of tsRNAs levels, mode/s of function, and identification of interacting proteins and RNAs may together shed light on the tsRNAs impact on neuronal function, and enable novel diagnostics/therapeutics avenues for brain diseases in age, sex and disease-specific manner.
    Keywords:  brain aging; neurodegeneration; neuroprotection; sex differences; tRNA mutations; tRNA-derived small RNAs (tsRNAs)
    DOI:  https://doi.org/10.1093/brain/awaf130
  38. Sci Rep. 2025 Apr 29. 15(1): 15078
    Attenuated PINK1 autophosphorylation play neuroprotective and anti-seizure roles in neonatal hypoxia.
    Yi Yuan, Xiaoqian Wang, Yaru Cui, Hua Zhou, Wenna Li, Qian Teng, Hongjin Wang, Bohan Sun, Qiaoyun Wang, Hongliu Sun, Jianhua Tang.
      This study investigated the roles and mechanisms of PINK1 activity in neonatal hypoxia-induced seizures with shRNA intervention targeting translocase outer mitochondrial membrane 7 (TOM7), the positive regulator of PINK1 autophosphorylation, or overlapping with the m-AAA protease 1 homolog (OMA1), the negative regulator of PINK1 autophosphorylation. Studies have suggested that in hypoxia-induced neonatal seizures, the phosphorylation level of PINK1 is significantly increased and the mitophagic pathway is activated, accompanied by neuronal damage and learning-memory deficits. Inhibiting PINK1 phosphorylation by reducing TOM7 expression alleviated mitophagy, mitochondrial oxidative stress, neuronal damage and seizures. In contrast, the inhibition of OMA1 expression resulted in a further increase in PINK1 phosphorylation and aggravated hypoxia-induced seizures and neuronal injury. This study implicated PINK1 activity in neonatal hypoxia and suggest that attenuated PINK1 autophosphorylation may have neuroprotective and anti-seizure effects in neonatal hypoxia.
    Keywords:  Mitochondrial oxidative stress; Mitophagy; Neuronal injury; PINK1; Seizure
    DOI:  https://doi.org/10.1038/s41598-025-99915-8
  39. Adv Healthc Mater. 2025 Apr 26. e2500301
    Organoid Vascularization: Strategies and Applications.
    Qianmin Gao, Jian Wang, Hao Zhang, Jianhua Wang, Yingying Jing, Jiacan Su.
      Organoids provide 3D structures that replicate native tissues in biomedical research. The development of vascular networks within organoids enables oxygen and nutrient delivery while facilitating metabolic waste removal, which supports organoid growth and maturation. Recent studies demonstrate that vascularized organoid models offer insights into tissue interactions and promote tissue regeneration. However, the current limitations in establishing functional vascular networks affect organoid growth, viability, and clinical translation potential. This review examines the development of vascularized organoids, including the mechanisms of angiogenesis and vasculogenesis, construction strategies, and biomedical applications. The approaches are categorized into in vivo and in vitro methods, with analysis of their specific advantages and limitations. The review also discusses emerging techniques such as bioprinting and gene editing for improving vascularization and functional integration in organoid-based therapies. Current developments in organoid vascularization indicate potential applications in modeling human diseases and developing therapeutic strategies, contributing to advances in translational research.
    Keywords:  angiogenesis; organoids; regenerative medicine; vascularization; vasculogenesis
    DOI:  https://doi.org/10.1002/adhm.202500301
  40. Eur J Pharmacol. 2025 Apr 24. pii: S0014-2999(25)00398-X. [Epub ahead of print] 177644
    Antisense Oligonucleotides: A Promising Advancement in Neurodegenerative Disease Treatment.
    Katarzyna Dudzisz, Ilona Wandzik.
      Antisense oligonucleotides (ASOs) are a class of therapeutics designed to modulate gene expression and have shown promise in the treatment of various neurodegenerative diseases. As of March 2025, four ASO-based therapies have received approval for the treatment of neurodegenerative diseases, including spinal muscular atrophy (SMA), amyotrophic lateral sclerosis (ALS), and hereditary transthyretin amyloidosis (ATTR). These approvals underscore the therapeutic potential of ASOs as effective treatments for neurodegenerative diseases by addressing specific genetic abnormalities. This is best demonstrated by clinical studies in more than a dozen ASOs, which could pave the way for the development of new therapeutics soon. Moreover, the ongoing extended clinical studies, which target presymptomatic carriers, have significant potential to cure familial ALS based on the SOD1 gene mutation. This review provides an update on clinical trials, highlighting promising results and the challenges encountered.
    Keywords:  antisense oligonucleotides; clinical trials; neurodegenerative diseases
    DOI:  https://doi.org/10.1016/j.ejphar.2025.177644
  41. Curr Protoc. 2025 Apr;5(4): e70136
    Purification of Low-Complexity Domain Proteins FUS, EWSR1, and Their Fusions.
    Jesse J Altemus, Michelle A Lay, Valery F Thompson, Jacob C Schwartz.
      FET proteins are large multifunctional proteins that have several key roles in biology. The FET family of proteins, including FUS, EWSR1, and TAF15, play critical roles in transcription regulation, RNA processing, and DNA damage repair. These multifunctional RNA- and DNA-binding proteins are ubiquitously expressed and conserved across vertebrate species. They contain low-complexity (LC) domains that allow them to assemble and phase separate but also makes the proteins prone to aggregation. Aberrations in FET proteins, such as point mutations, aggregation, or translocations leading to fusion proteins, have been implicated in several pathologies, including frontotemporal lobar degeneration (FTLD), amyotrophic lateral sclerosis (ALS), and Ewing sarcoma. In vitro study of FET proteins is hampered by their propensity to aggregate, their disordered structure, and their susceptibility to proteolysis, making high-yield production difficult. Here, we present optimized methods for the purification of full-length FUS, EWSR1, and their fusion proteins. These protocols enable researchers to overcome issues related to aggregation and solubility, facilitating biochemical and biophysical studies of these critical yet complex proteins. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol: Purification of EWSR1 and FUS proteins Alternate Protocol: Purification for fusion proteins.
    Keywords:  Ewing sarcoma; fusion protein; low‐complexity domain; neurodegenerative disease; phase separation; protein purification
    DOI:  https://doi.org/10.1002/cpz1.70136
  42. NeuroImmune Pharm Ther. 2025 Mar;4(1): 1-11
    Oligonucleotide therapeutics for neurodegenerative diseases.
    Victor Li, Yunlong Huang.
      Recently there has been a surge in interest involving the application of oligonucleotides, including small interfering RNA (siRNA) and antisense oligonucleotides (ASOs), for the treatment of chronic diseases that have few available therapeutic options. This emerging class of drugs primarily operates by selectively suppressing target genes through antisense and/or RNA interference mechanisms. While various commercial medications exist for delivering oligonucleotides to the hepatic tissue, achieving effective delivery to extra hepatic tissues remains a formidable challenge. Here, we review recent advances in oligonucleotide technologies, including nanoparticle delivery, local administration, and 2'-O-hexadecyl (C16)-conjugation that work to extend the applicability of siRNAs and ASOs to nerve tissues. We discuss critical factors pivotal for the successful clinical translations of these modified or engineered oligonucleotides in the context of treating neurodegenerative diseases such as Alzheimer's disease and amyotrophic lateral sclerosis.
    Keywords:  Alzheimer’s disease; amyotrophic lateral sclerosis; antisense oligonucleotides; oligonucleotide therapeutics; small interfering RNA
    DOI:  https://doi.org/10.1515/nipt-2024-0013
  43. Methods Mol Biol. 2025 ;2924 223-233
    Engineered Hydrogels for 3D Cell Culture and Bioprinting of Human Induced Pluripotent Stem Cell-Derived Neuroepithelial Stem Cells.
    Daniel Aili, Anna Herland.
      This protocol outlines the synthesis and use of engineered hyaluronan-based hydrogels for 3D cell culture and bioprinting of human induced pluripotent stem cell (hiPSC)-derived neuroepithelial stem cells (lt-NES). Key steps include hydrogel formation using bioorthogonal chemistries, cell encapsulation, and 3D bioprinting with a Cellink BioX printer, enabling the creation of complex tissue models. The protocol ensures high cell viability and supports differentiation, essential for neuroscience research and drug development.
    Keywords:  3D bioprinting; 3D cell culture; Hyaluronan; Hydrogels; Laminin; Neuroepithelial stem cells
    DOI:  https://doi.org/10.1007/978-1-0716-4530-7_16
  44. Alzheimers Dement. 2025 Apr;21(4): e70208
    A comparison of p-tau assays for the specificity to detect tau changes in Alzheimer's disease.
    Shorena Janelidze, Nicholas J Ashton, Anna Orduña Dolado, Ulrika Nordström, Divya Bali, Karin M E Forsberg, Isil Keskin, Andrea Mastrangelo, Veria Vacchiano, Rocco Liguori, Kaj Blennow, Henrik Zetterberg, Niklas Mattsson-Carlgren, Fernando Gonzalez-Ortiz, Piero Parchi, Peter Munch Andersen, Oskar Hansson.
       INTRODUCTION: We evaluated differences in p-tau levels between Alzheimer's disease (AD), a condition with brain-specific changes in p-tau, and amyotrophic lateral sclerosis (ALS), a condition associated with increases in peripheral p-tau levels.
    METHODS: Cerebrospinal fluid and plasma from 668 participants were analyzed using immunoassays specific for the low-molecular-weight (LMW) tau isoforms present in the brain (i.e., p-tau217Lilly, p-tau181Lilly) and those that detect both LMW- and high-molecular-weight (HMW) tau expressed in the peripheral nervous system (i.e., p-tau217AlzPath, p-tau181UGOT).
    RESULTS: Increases in plasma p-tau in ALS versus controls were significantly smaller for the LMW-specific p-tau assays (15.9%-20.5%) compared with non-specific assays (92.0%-121.3%). The LMW-specific p-tau assays showed significantly larger plasma p-tau increases in AD versus ALS, discriminating AD from ALS with areas under the curve (AUCs; 0.890.93) higher than the AUCs of the non-specific assays (0.54-0.74).
    DISCUSSION: LMW-specific p-tau assays could be more useful in the diagnostic workup of AD, especially in population-based communities where conditions causing peripheral neuropathy are frequent.
    HIGHLIGHTS: Increases in plasma phosphorylated tau (p-tau) in amyotrophic lateral sclerosis (ALS) versus controls were significantly smaller for low-molecular-weight (LMW)-specific p-tau assays (i.e., p-tau217Lilly, p-tau181Lilly) compared with p-tau assays that also detect high-molecular-weight (HMW) assays (i.e., p-tau217AlzPath, p-tau181UGOT). The LMW-specific p-tau assays showed significantly larger increases in plasma p-tau in AD versus ALS compared with the non-specific assays. The LMW-specific p-tau assays discriminated AD from ALS with higher precision, showing significantly better performance than the non-specific assays. LMW-specific p-tau assays could be more useful in the diagnostic workup of AD, especially in population-based communities where conditions causing peripheral neuropathy (such as ALS) are frequent.
    Keywords:  Alzheimer's disease; amyotrophic lateral sclerosis; biomarker; blood; low‐molecular‐weight tau; p‐tau
    DOI:  https://doi.org/10.1002/alz.70208
  45. Methods Mol Biol. 2025 ;2924 1-14
    Cell-Based Assays Using Derived Human-Induced Pluripotent Cells in Drug Discovery and Development.
    James A Ross, Carl-Fredrik Mandenius.
      The use of human-induced pluripotent cell lines in the differentiation of specific cell types and assay development has in recent years progressed significantly. In this introduction to the field, the rationale for using induced pluripotent stem cells in drug development is described and some of the advantages and drawbacks are discussed. The prospect of facilitating drug development by using stratified sources of cells from diverse patient groups and differentiating these to organ and tissue cells for their use in disease models, screening assays, and ADME testing is highlighted. The variety of methodologies and protocols presented in this volume provides a selected collection of experiences and practice. Here, these are discussed in a general perspective with references and comments to other similar protocols in the literature.
    Keywords:  Differentiation protocols; Disease models; Drug screening; Induced pluripotent cell; iPSC
    DOI:  https://doi.org/10.1007/978-1-0716-4530-7_1
  46. Neuropharmacology. 2025 Apr 30. pii: S0028-3908(25)00190-X. [Epub ahead of print] 110484
    SIRTUINS MODULATORS MITIGATE HYPOXIA-INDUCED CELL DEATH DUE TO CHANGES IN HISTONE 3 ACETYLATION, AND MITOCHONDRIAL FUNCTION, DYNAMICS, AND CONTENT.
    Luiz Felipe Souza E Silva, Amanda Siena, Jessica Mayumi Yuzawa, Tatiana Rosado Rosenstock.
      Hypoxia is a key environmental factor linked to neurodevelopmental complications, primarily through its impact on mitochondrial dysfunction. Given that sirtuins regulate mitochondrial and cellular metabolism, we aimed to investigate whether pharmacological modulation of sirtuins could protect neurons from hypoxia-induced mitochondrial dysfunction and cell death. To explore this, primary cortical neurons from male Wistar rats (control) and Spontaneously Hypertensive Rats (a model for neonatal hypoxia and schizophrenia) were exposed to cobalt chloride (CoCl2) to chemically induce hypoxia. Neurons were also treated with Nicotinamide (50 μM), Resveratrol (0.5 μM), and Sirtinol (5 μM) to modulate sirtuin activity. We first assessed histone deacetylation, cell death, mitochondrial calcium retention capacity, mitochondrial membrane potential, and levels of reactive oxygen species (ROS). In addition, we analysed the expression of genes related to mitochondrial metabolism, dynamics, and biogenesis, as well as high-energy compound levels. Our data indicate that both chemical and neonatal hypoxia caused mitochondrial depolarization, reduced calcium retention, increased ROS levels, and elevated Nfe2l2 expression in primary cortical neurons. Hypoxia also led to increased expression of genes associated with mitochondrial biogenesis and fission, as well as reduced ATP levels and elevated pyruvate and lactate levels. Importantly, treatment with sirtuin modulators enhanced neuron viability, likely by further increasing Nfe2l2 expression and reducing ROS production. These modulators also improved metabolic outcomes, including higher ATP levels, and normalized pyruvate and lactate production, as well as mitochondrial fusion gene expression. Collectively, our findings suggest that sirtuin modulators could mitigate hypoxia-induced damage and may represent a potential therapeutic strategy for managing neurodevelopmental disorders.
    Keywords:  Hypoxia; Mitochondrial dysfunction; Schizophrenia and Neurons; Sirtuins
    DOI:  https://doi.org/10.1016/j.neuropharm.2025.110484
  47. Biology (Basel). 2025 Apr 14. pii: 420. [Epub ahead of print]14(4):
    A Novel Approach to Relocate Misplaced Proteins in Cells.
    Grace Hohman, Ava Watson, Mohamed A Eldeeb.
      Proper cellular function hinges on appropriate subcellular protein localization. When cellular proteins become mislocalized, they can accumulate, cause cellular damage, and disrupt many biochemical and cellular processes. Notably, mislocalized protein accumulation and the resulting cytotoxic effects are salient features of neurodegenerative diseases including Alzheimer's, Parkinson's disease, and ALS. The detrimental cellular consequences of mislocalized proteins accumulation make it crucial to develop techniques and approaches that counteract this malfunction. Remarkably, a recent study by Ng et al. introduced targeted relocalization-activating molecules (TRAMs) as a novel molecular tool for relocalizing endogenous target proteins to counteract disease-associated mislocalized proteins. The authors developed a quantitative single-cell analysis to evaluate the strength and relocalization capability of TRAMs by coupling a target protein and a shuttle protein. Herein, we briefly highlight and discuss the potential molecular implications for targeted protein relocalization as an effective approach for correcting mislocalized proteins.
    Keywords:  misfolded proteins; neurodegenerative disease; protein localization; protein quality control; protein targeting
    DOI:  https://doi.org/10.3390/biology14040420
  48. Aging Cell. 2025 Apr 25. e70054
    3D Mitochondrial Structure in Aging Human Skeletal Muscle: Insights Into MFN-2-Mediated Changes.
    Estevão Scudese, Andrea G Marshall, Zer Vue, Vernat Exil, Benjamin I Rodriguez, Mert Demirci, Larry Vang, Edgar Garza López, Kit Neikirk, Bryanna Shao, Han Le, Dominique Stephens, Duane D Hall, Rahmati Rostami, Taylor Rodman, Kinuthia Kabugi, Jian-Qiang Shao, Margaret Mungai, Salma T AshShareef, Innes Hicsasmaz, Sasha Manus, Celestine N Wanjalla, Aaron Whiteside, Revathi Dasari, Clintoria R Williams, Steven M Damo, Jennifer A Gaddy, Brian Glancy, Estélio Henrique Martin Dantas, André Kinder, Ashlesha Kadam, Dhanendra Tomar, Fabiana Scartoni, Matheus Baffi, Melanie R McReynolds, Mark A Phillips, Anthonya Cooper, Sandra A Murray, Anita M Quintana, Nelson Wandira, Okwute M Ochayi, Magdalene Ameka, Annet Kirabo, Sepiso K Masenga, Chanel Harris, Ashton Oliver, Pamela Martin, Amadou Gaye, Olga Korolkova, Vineeta Sharma, Bret C Mobley, Prasanna Katti, Antentor Hinton.
      Age-related skeletal muscle atrophy, known as sarcopenia, is characterized by loss of muscle mass, strength, endurance, and oxidative capacity. Although exercise has been shown to mitigate sarcopenia, the underlying governing mechanisms are poorly understood. Mitochondrial dysfunction is implicated in aging and sarcopenia; however, few studies explore how mitochondrial structure contributes to this dysfunction. In this study, we sought to understand how aging impacts mitochondrial three-dimensional (3D) structure and its regulators in skeletal muscle. We hypothesized that aging leads to remodeling of mitochondrial 3D architecture permissive to dysfunction and is ameliorated by exercise. Using serial block-face scanning electron microscopy (SBF-SEM) and Amira software, mitochondrial 3D reconstructions from patient biopsies were generated and analyzed. Across five human cohorts, we correlate differences in magnetic resonance imaging, mitochondria 3D structure, exercise parameters, and plasma immune markers between young (under 50 years) and old (over 50 years) individuals. We found that mitochondria are less spherical and more complex, indicating age-related declines in contact site capacity. Additionally, aged samples showed a larger volume phenotype in both female and male humans, indicating potential mitochondrial swelling. Concomitantly, muscle area, exercise capacity, and mitochondrial dynamic proteins showed age-related losses. Exercise stimulation restored mitofusin 2 (MFN2), one such of these mitochondrial dynamic proteins, which we show is required for the integrity of mitochondrial structure. Furthermore, we show that this pathway is evolutionarily conserved, as Marf, the MFN2 ortholog in Drosophila, knockdown alters mitochondrial morphology and leads to the downregulation of genes regulating mitochondrial processes. Our results define age-related structural changes in mitochondria and further suggest that exercise may mitigate age-related structural decline through modulation of mitofusin 2.
    Keywords:  3D reconstruction; MFN‐2; aging; exercise; human skeletal muscle; mitochondria
    DOI:  https://doi.org/10.1111/acel.70054
  49. Sci Signal. 2025 Apr 29. 18(884): eady4818
    A good side of STING.
    Leslie K Ferrarelli.
      The innate immunity mediator STING senses and repairs lysosomal dysfunction.
    DOI:  https://doi.org/10.1126/scisignal.ady4818
  50. Eur J Clin Invest. 2025 Apr 25. e70051
    Extracellular matrix mechanical cues (dys)regulate metabolic redox homeostasis due to impaired autophagic flux.
    Heloísa Gerardo, Tânia Lourenço, Júlio Torres, Manuela Ferreira, Célia Aveleira, Susana Simões, Lino Ferreira, Cláudia Cavadas, Paulo J Oliveira, José Teixeira, Mário Grãos.
       BACKGROUND: Extracellular matrix (ECM) stiffness is increasingly recognized as a critical regulator of cellular behaviour, governing processes such as proliferation, differentiation, and metabolism. Neurodegenerative diseases are characterized by mitochondrial dysfunction, oxidative stress, impaired autophagy, and progressive softening of the brain tissue, yet research into how mechanical cues influence cellular metabolism in this context remains scarce.
    MATERIALS AND METHODS: In this study, we evaluated the long-term effects of brain-compliant, soft ECM on mitochondrial bioenergetics, redox balance, and autophagic capacity in human neuroblastoma (SH-SY5Y) and mouse hippocampal (HT22) cell lines, as well as primary mouse neurons.
    RESULTS: We observed that prolonged exposure to soft ECM does not impact cell proliferative capacity of neuronal cells but results in mitochondrial bioenergetic dysfunction, redox imbalance, and disrupted autophagic flux. These findings were consistently validated across both human and mouse neuronal cells. Our data indicate a decreased maximal autophagic capacity in cells exposed to long-term soft ECM, potentially due to an imbalance in autophagosome formation and degradation, as demonstrated by decreased LC3 II levels following chloroquine-induced autophagic flux inhibition. This impairment in autophagy was coupled with increased cellular oxidative stress, further indicating metabolic alterations.
    CONCLUSIONS: These findings emphasize the critical role of ECM stiffness in regulating neuronal cell metabolism and suggest that prolonged exposure to soft ECM may mimic key aspects of neurodegenerative disease pathology, thereby enhancing the physiological relevance of in vitro models. This study underscores the necessity for further research into ECM mechanics as a contributing factor in neurodegenerative disease progression and as a potential target for therapeutic strategies.
    Keywords:  autophagy; extracellular matrix; mechanical cues; mitochondria bioenergetic; neuronal cells; redox homeostasis
    DOI:  https://doi.org/10.1111/eci.70051
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