Revisiting astrocyte to neuron conversion with lineage tracing in vivo

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 precis...

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Published in:	Cell Vol. 184; no. 21; p. 5465
Main Authors:	Wang, Lei-Lei, Serrano, Carolina, Zhong, Xiaoling, Ma, Shuaipeng, Zou, Yuhua, Zhang, Chun-Li
Format:	Journal Article
Language:	English
Published:	United States 14.10.2021
Subjects:	Animals Astrocytes - cytology Astrocytes - metabolism Basic Helix-Loop-Helix Transcription Factors - metabolism Brain - pathology Brain Injuries - pathology Cell Differentiation Cell Line, Tumor Cell Lineage Cellular Reprogramming Dependovirus - metabolism Down-Regulation Gene Expression Regulation Genes, Reporter Glial Fibrillary Acidic Protein - genetics Heterogeneous-Nuclear Ribonucleoproteins - metabolism Homeodomain Proteins - metabolism Humans Integrases - metabolism Mice Mice, Inbred C57BL Mice, Transgenic Neurons - cytology Neurons - metabolism Polypyrimidine Tract-Binding Protein - metabolism Promoter Regions, Genetic - genetics Transcription Factors - metabolism NEUROD1 in vivo reprogramming PTBP1 DLX2 astrocyte-to-neuron conversion shRNA AAV PAX6 lineage tracing CRISPR-CasRx
ISSN:	1097-4172, 1097-4172
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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.
AbstractList	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. 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.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.
Author	Serrano, Carolina Ma, Shuaipeng Zou, Yuhua Wang, Lei-Lei Zhong, Xiaoling Zhang, Chun-Li
Author_xml	– sequence: 1 givenname: Lei-Lei surname: Wang fullname: Wang, Lei-Lei email: leilei.wang@utsouthwestern.edu organization: Department of Molecular Biology and Hamon Center for Regenerative Science and Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA. Electronic address: leilei.wang@utsouthwestern.edu – sequence: 2 givenname: Carolina surname: Serrano fullname: Serrano, Carolina organization: Department of Molecular Biology and Hamon Center for Regenerative Science and Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA – sequence: 3 givenname: Xiaoling surname: Zhong fullname: Zhong, Xiaoling organization: Department of Molecular Biology and Hamon Center for Regenerative Science and Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA – sequence: 4 givenname: Shuaipeng surname: Ma fullname: Ma, Shuaipeng organization: Department of Molecular Biology and Hamon Center for Regenerative Science and Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA – sequence: 5 givenname: Yuhua surname: Zou fullname: Zou, Yuhua organization: Department of Molecular Biology and Hamon Center for Regenerative Science and Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA – sequence: 6 givenname: Chun-Li surname: Zhang fullname: Zhang, Chun-Li email: chun-li.zhang@utsouthwestern.edu organization: Department of Molecular Biology and Hamon Center for Regenerative Science and Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA. Electronic address: chun-li.zhang@utsouthwestern.edu
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References	34653366 - Cell. 2021 Oct 14;184(21):5303-5305 34699780 - Mol Ther. 2021 Nov 3;29(11):3097-3098 35108505 - Mol Ther. 2022 Mar 2;30(3):986-987 35123657 - Mol Ther. 2022 Mar 2;30(3):982-986
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Snippet	In vivo cell fate conversions have emerged as potential regeneration-based therapeutics for injury and disease. Recent studies reported that ectopic expression... In vivo cell fate conversions have emerged as potential regeneration-based therapeutics for injury and disease. Recent studies reported that ectopic expression...
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SubjectTerms	Animals Astrocytes - cytology Astrocytes - metabolism Basic Helix-Loop-Helix Transcription Factors - metabolism Brain - pathology Brain Injuries - pathology Cell Differentiation Cell Line, Tumor Cell Lineage Cellular Reprogramming Dependovirus - metabolism Down-Regulation Gene Expression Regulation Genes, Reporter Glial Fibrillary Acidic Protein - genetics Heterogeneous-Nuclear Ribonucleoproteins - metabolism Homeodomain Proteins - metabolism Humans Integrases - metabolism Mice Mice, Inbred C57BL Mice, Transgenic Neurons - cytology Neurons - metabolism Polypyrimidine Tract-Binding Protein - metabolism Promoter Regions, Genetic - genetics Transcription Factors - metabolism
Title	Revisiting astrocyte to neuron conversion with lineage tracing in vivo
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