Micro-RNAs secreted through astrocyte-derived extracellular vesicles cause neuronal network degeneration in C9orf72 ALS
Open accessAbstract
Background Astrocytes regulate neuronal function, synaptic formation and maintenance partly through secreted extracellular vesicles (EVs). In amyotrophic lateral sclerosis (ALS) astrocytes display a toxic phenotype that contributes to motor neuron (MN) degeneration. Methods We used human induced astrocytes (iAstrocytes) from 3 ALS patients carrying C9orf72 mutations and 3 non-affected donors to investigate the role of astrocyte-derived EVs (ADEVs) in ALS astrocyte toxicity. ADEVs were isolated from iAstrocyte conditioned medium via ultracentrifugation and resuspended in fresh astrocyte medium before testing ADEV impact on HB9-GFP + mouse motor neurons (Hb9-GFP + MN). We used post-mortem brain and spinal cord tissue from 3 sporadic ALS and 3 non-ALS cases for PCR analysis. Findings We report that EV formation and miRNA cargo are dysregulated in C9ORF72-ALS iAstrocytes and this affects neurite network maintenance and MN survival in vitro . In particular, we have identified downregulation of miR-494-3p, a negative regulator of semaphorin 3A (SEMA3A) and other targets involved in axonal maintenance. We show here that by restoring miR-494-3p levels through expression of an engineered miRNA mimic we can downregulate Sema3A levels in MNs and increases MN survival in vitro . Consistently, we also report lower levels of mir-494-3p in cortico-spinal tract tissue isolated from sporadic ALS donors, thus supporting the pathological importance of this pathway in MNs and its therapeutic potential. Interpretation ALS ADEVs and their miRNA cargo are involved in MN death in ALS and we have identified miR-494-3p as a potential therapeutic target. Funding: Thierry Latran Fondation and Academy of Medical Sciences.
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Astrocytes generated from hiPSC derived from fibroblasts of patients with CNS diseases show pathological phenotypes, including dysregulation of lipid metabolism, alteration in extracellular vesicles, reduced autophagy, or altered STAT3 signaling.
“Interestingly, astrocytes generated from hiPSC derived from fibroblasts obtained from patients with CNS diseases (usually with a genetic mutation causative of disease or a risk polymorphism) show pathological phenotypes, including dysregulation of lipid metabolism11, alteration in the contents of the extracellular vesicles released by astrocytes80, reduced autophagy81, or altered STAT3 signaling82”
From Reactive astrocyte nomenclature, definitions, and future directions (2021) · cites this paper as [80]
The Results section (“MicroRNAs regulating axonal maintenance are selectively dysregulated in C9 patient ADEVs”; page unavailable) reports altered miRNA cargo in extracellular vesicles released by C9orf72-ALS patient astrocytes, supporting the point attached to this citation. However, the Materials and methods section (“Conversion of skin fibroblasts to induced neural progenitor cells”; page unavailable) describes direct conversion of patient fibroblasts to neural progenitor cells and then astrocytes, not generation through hiPSCs. (LLM, conf=0.88)
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- Authors
- Varcianna, A., Myszczynska, M. A., Castelli, L. M., O'Neill, B., Kim, Y., Talbot, J., Nyberg, S., Nyamali, I., Heath, P. R., Stopford, M. J., Hautbergue, G. M., & Ferraiuolo, L.
- Affiliations
- University of Sheffield (GB)
- Corresponding author
- Ferraiuolo, Laura
- ORCID
- Venue
- EBioMedicine
- Publisher
- Elsevier BV
- Published
- 2019-02
- Volume
- 40
- Pages
- 626-635
- ISSN
- 2352-3964 (ISSN-L)
- PMC
- PMC6413467
- OpenAlex
- W2911503339
- Cited by
- 194
- References
- 45
- Citations by year
- 2026: 14 · 2025: 13 · 2024: 32 · 2023: 32 · 2022: 34 · 2021: 39 · 2020: 23 · 2019: 7
- FWCI
- 17.43
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- EBioMedicine (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- White Rose Research Online (University of Leeds, The University of Sheffield, University of York) (acceptedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Wellcome Trust
- Motor Neurone Disease Association (Bonanno/Apr16/848-791, Hautbergue/Apr16/846-791)
- Academy of Medical Sciences (SBF002\1142)
- Medical Research Council (MR/R024162/1, 1812144)
- Topics
- Health Sciences › Medicine › Neurology › Amyotrophic Lateral Sclerosis Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › MicroRNA in disease regulation
- Keywords
- amyotrophic lateral sclerosis, C9orf72, microRNA, miR-494-3p, semaphorin 3A, motor neuron degeneration, astrocyte toxicity, corticospinal tract, therapeutic target
- MeSH terms
- Extracellular Vesicles, C9orf72 Protein, Adult, Aged, Amyotrophic Lateral Sclerosis, Animals, Astrocytes, Autopsy, Biopsy, Cell Survival, Disease Models, Animal, Disease Susceptibility, Female, Gene Expression Regulation, Humans, Male, Middle Aged, Models, Biological, Motor Neurons, Mutation, Nerve Degeneration, Skin, Culture Media, Conditioned, Gene Expression Profiling, RNA Interference, MicroRNAs, Semaphorin-3A, Mice
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