Revisiting astrocyte to neuron conversion with lineage tracing in vivo
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Abstract
In vivo cell fate conversions have emerged as potential regeneration-based therapeutics for injury and disease. Recent studies reported that ectopic expression or knockdown of certain factors can convert resident astrocytes into functional neurons with high efficiency, region specificity, and precise connectivity. However, using stringent lineage tracing in the mouse brain, we show that the presumed astrocyte-converted neurons are actually endogenous neurons. AAV-mediated co-expression of NEUROD1 and a reporter specifically and efficiently induces reporter-labeled neurons. However, these neurons cannot be traced retrospectively to quiescent or reactive astrocytes using lineage-mapping strategies. Instead, through a retrograde labeling approach, our results reveal that endogenous neurons are the source for these viral-reporter-labeled neurons. Similarly, despite efficient knockdown of PTBP1 in vivo, genetically traced resident astrocytes were not converted into neurons. Together, our results highlight the requirement of lineage-tracing strategies, which should be broadly applied to studies of cell fate conversions in vivo.
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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 [16]
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 [16]
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- Authors
- Wang, L.-L., Serrano, C., Zhong, X., Ma, S., Zou, Y., & Zhang, C.-L.
- Affiliations
- The University of Texas Southwestern Medical Center (US)
- Corresponding author
- Wang, Lei-Lei
- Zhang, Chun-Li
- Venue
- Cell
- Publisher
- Elsevier BV
- Published
- 2021-10
- Volume
- 184
- Issue
- 21
- Pages
- 5465-5481.e16
- ISSN
- 0092-8674 (ISSN-L)
- PMC
- PMC8526404
- OpenAlex
- W3203538421
- Cited by
- 329
- References
- 65
- Citations by year
- 2026: 30 · 2025: 60 · 2024: 74 · 2023: 68 · 2022: 73 · 2021: 21 · 2020: 3
- FWCI
- 21.49
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- www.elsevier.com/tdm/userlicense/1.0/ (tdm)
- www.elsevier.com/legal/tdmrep-license (tdm)
- www.elsevier.com/open-access/userlicense/1.0/ (vor, 378 d)
- doi.org/10.15223/policy-017 (stm-asf)
- doi.org/10.15223/policy-037 (stm-asf)
- doi.org/10.15223/policy-012 (stm-asf)
- doi.org/10.15223/policy-029 (stm-asf)
- doi.org/10.15223/policy-004 (stm-asf)
- Locations
- Cell (publishedVersion)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- Welch Foundation (I-1724)
- National Institutes of Health (NS099073, NS117065, NS111776, NS092616, NS088095)
- Topics
- Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
- Keywords
- astrocyte-to-neuron conversion, lineage tracing, retrograde tracing, NeuroD1, PTBP1, reactive astrocytes, cell fate conversion, brain regeneration, mouse brain
- MeSH terms
- Dependovirus, Animals, Astrocytes, Brain, Brain Injuries, Cell Differentiation, Gene Expression Regulation, Glial Fibrillary Acidic Protein, Humans, Mice, Inbred C57BL, Mice, Transgenic, Neurons, Promoter Regions, Genetic, Transcription Factors, Down-Regulation, Genes, Reporter, Homeodomain Proteins, Cell Lineage, Integrases, Heterogeneous-Nuclear Ribonucleoproteins, Polypyrimidine Tract-Binding Protein, Cell Line, Tumor, Mice, Basic Helix-Loop-Helix Transcription Factors, Basic Helix-Loop-Helix Proteins, Cellular Reprogramming
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