Repressing PTBP1 fails to convert reactive astrocytes to dopaminergic neurons in a 6-hydroxydopamine mouse model of Parkinson's disease
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Abstract
Lineage reprogramming of resident glial cells to dopaminergic neurons (DAns) is an attractive prospect of the cell-replacement therapy for Parkinson’s disease (PD). However, it is unclear whether repressing polypyrimidine tract binding protein 1 (PTBP1) could efficiently convert astrocyte to DAns in the substantia nigra and striatum. Although reporter-positive DAns were observed in both groups after delivering the adeno-associated virus (AAV) expressing a reporter with shRNA or CRISPR-CasRx to repress astroglial PTBP1, the possibility of AAV leaking into endogenous DAns could not be excluded without using a reliable lineage-tracing method. By adopting stringent lineage-tracing strategy, two other studies show that either knockdown or genetic deletion of quiescent astroglial PTBP1 fails to obtain induced DAns under physiological condition. However, the role of reactive astrocytes might be underestimated because upon brain injury, reactive astrocyte can acquire certain stem cell hallmarks that may facilitate the lineage conversion process. Therefore, whether reactive astrocytes could be genuinely converted to DAns after PTBP1 repression in a PD model needs further validation. In this study, we used Aldh1l1-CreERT2 -mediated specific astrocyte-lineage-tracing method to investigate whether reactive astrocytes could be converted to DAns in a 6-hydroxydopamine (6-OHDA) mouse model of PD. However, we found that no astrocyte-originated DAn was generated after effective and persistent knockdown of astroglial PTBP1 either in the substantia nigra or in striatum, while AAV ‘leakage’ to nearby neurons was easily observed. Our results confirm that repressing PTBP1 does not convert astrocytes to DAns, regardless of physiological or PD-related pathological conditions.
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Some reported conversions reflected viral promoter leakage into pre-existing neurons.
“However, subsequent lineage-tracing studies yielded conflicting results, suggesting that PTBP1 depletion alone may be insufficient to drive genuine astrocyte-to-neuron conversion in the adult mammalian brain, raising concerns that some reported conversions reflected viral promoter leakage into pre-existing neurons.”
From Reverse the progression of Alzheimer’s disease through Nano-ERASER-based adult neuroregeneration (2026) · cites this paper as [17]
Subsequent lineage-tracing studies yielded conflicting results, suggesting that PTBP1 depletion alone may be insufficient to drive genuine astrocyte-to-neuron conversion in the adult mammalian brain.
“However, subsequent lineage-tracing studies yielded conflicting results, suggesting that PTBP1 depletion alone may be insufficient to drive genuine astrocyte-to-neuron conversion in the adult mammalian brain, raising concerns that some reported conversions reflected viral promoter leakage into pre-existing neurons.”
From Reverse the progression of Alzheimer’s disease through Nano-ERASER-based adult neuroregeneration (2026) · cites this paper as [17]
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Ptbp1 knockdown failed to induce astrocytes to neurons in vivo
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PTBP1 regulates injury responses and sensory pathways in adult peripheral neurons
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Therapeutic Potential of PTB Inhibition Through Converting Glial Cells to Neurons in the Brain
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Role of astrocytes in the pathogenesis of perinatal brain injury
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Develop an efficient and specific AAV-based labeling system for Muller glia in mice
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Post-transcriptional mechanisms controlling neurogenesis and direct neuronal reprogramming
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Post-transcriptional regulation of the transcriptional apparatus in neuronal development
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Base editing of Ptbp1 in neurons alleviates symptoms in a mouse model of Parkinson’s diseasePreprint
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Strategies and mechanisms of neuronal reprogramming
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The role of transcriptional and epigenetic modifications in astrogliogenesis
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A cutting-edge strategy for spinal cord injury treatment: resident cellular transdifferentiation
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Delivery Systems in Neuronal Direct Cell Reprogramming
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RNA therapeutics for neurological disease
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PTBP1 depletion in mature astrocytes reveals distinct splicing alterations without neuronal featuresPreprint
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Ptbp1 is not required for retinal neurogenesis and cell fate specification
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PTBP1 Depletion in Mature Astrocytes Reveals Distinct Splicing Alterations Without Neuronal FeaturesPreprint
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Ptbp1 is not required for retinal neurogenesis and cell fate specificationPreprint
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RNA Splicing Dysregulation in Parkinson’s Disease
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- Authors
- Chen, W., Zheng, Q., Huang, Q., Ma, S., & Li, M.
- Affiliations
- Sun Yat-sen University (CN)
- Corresponding author
- Ma, Shanshan
- Li, Mingtao
- Venue
- eLife
- Publisher
- eLife Sciences Publications, Ltd
- Published
- 2022-05-10
- Volume
- 11
- ISSN
- 2050-084X (ISSN-L)
- PMC
- PMC9208759
- OpenAlex
- W4229450008
- Cited by
- 89
- References
- 69
- Citations by year
- 2026: 13 · 2025: 16 · 2024: 23 · 2023: 29 · 2022: 8
- FWCI
- 6.20
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- eLife (publishedVersion, cc-by)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Ministry of Science and Technology of China (the National Key R&D Program of China (2018YFA0108300))
- National Natural Science Foundation of China (U1801681, 81771368, 31871019, 32070959)
- Department of Science and Technology of Guangdong Province (the Key Realm R&D Program of Guangdong Province (2018B030337001), the Guangdong Provincial Key Laboratory of Brain Function and Disease (2020B1212060024))
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Health Sciences › Medicine › Neurology › Parkinson's Disease Mechanisms and Treatments
- Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
- Keywords
- PTBP1, reactive astrocytes, Parkinson's disease, lineage reprogramming, substantia nigra, striatum, AAV gene delivery, shRNA knockdown, lineage tracing, dopaminergic neurons
- MeSH terms
- Dependovirus, Animals, Astrocytes, Disease Models, Animal, Parkinson Disease, Substantia Nigra, Oxidopamine, Heterogeneous-Nuclear Ribonucleoproteins, Polypyrimidine Tract-Binding Protein, Mice, Dopaminergic Neurons
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