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A NeuroD1 AAV-Based Gene Therapy For Functional Brain Repair After Ischemic Injury Through In Vivo Astrocyte-To-Neuron Conversion

ABSTRACT: A NeuroD1 AAV-Based Gene Therapy For Functional Brain Repair After Ischemic Injury Through In Vivo Astrocyte-To-Neuron Conversion

PROVIDER: PRJNA560878 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR10001352.fastq.gz	Fastqsanger.gz
	SRR10001353.fastq.gz	Fastqsanger.gz
	SRR10001354.fastq.gz	Fastqsanger.gz
	SRR10001355.fastq.gz	Fastqsanger.gz
	SRR10001356.fastq.gz	Fastqsanger.gz

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A NeuroD1 AAV-Based Gene Therapy For Functional Brain Repair After Ischemic Injury Through In Vivo Astrocyte-To-Neuron Conversion

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Project description:Direct astrocyte-to-neuron conversion holds promise for in situ neuroregeneration to treat neurodegenerative diseases. However, commonly-used adeno-associated vectors carrying GFAP mini-promoters drive leaky expression of neurogenic factors in endogenous neurons, raising concerns about trans-differentiation validity. Developing inducible, lineage-traceable transgenic mice, we evaluate NeuroD1 effects on astrocyte-to-neuron conversion in spinal cord and brain injury models. Lineage-tracing and single-cell transcriptomics reveal that ectopic NeuroD1 expression promotes robust conversion of reactive astrocytes in lesion cores to neurons at the acute phase of injury, but not in peri-injury astrocytes.

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A NeuroD1 AAV-Based Gene Therapy for Functional Brain Repair after Ischemic Injury through In Vivo Astrocyte-to-Neuron Conversion.

Project description:Adult mammalian brains have largely lost neuroregeneration capability except for a few niches. Previous studies have converted glial cells into neurons, but the total number of neurons generated is limited and the therapeutic potential is unclear. Here, we demonstrate that NeuroD1-mediated in situ astrocyte-to-neuron conversion can regenerate a large number of functional new neurons after ischemic injury. Specifically, using NeuroD1 adeno-associated virus (AAV)-based gene therapy, we were able to regenerate one third of the total lost neurons caused by ischemic injury and simultaneously protect another one third of injured neurons, leading to a significant neuronal recovery. RNA sequencing and immunostaining confirmed neuronal recovery after cell conversion at both the mRNA level and protein level. Brain slice recordings found that the astrocyte-converted neurons showed robust action potentials and synaptic responses at 2 months after NeuroD1 expression. Anterograde and retrograde tracing revealed long-range axonal projections from astrocyte-converted neurons to their target regions in a time-dependent manner. Behavioral analyses showed a significant improvement of both motor and cognitive functions after cell conversion. Together, these results demonstrate that in vivo cell conversion technology through NeuroD1-based gene therapy can regenerate a large number of functional new neurons to restore lost neuronal functions after injury.

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