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Predicting cellular reprogramming for cell replacement

Reference number
SB16-0065
Start and end dates
170801-230731
Amount granted
34 000 000 SEK
Administrative organization
Karolinska Institutet
Research area
Life Sciences

Summary

In neurodegenerative disease such as Parkinson’s, it may be possible to replace lost dopamineergic neurons by new cells derived from the patient’s own endogenous brain astrocytes. This requires reprogramming, which in turn requires a mechanistic understanding of how dopaminergic neurons are specified and maintained. To achieve these goals, we will 1. Create a single-cell atlas of human embryonic ventral midbrain 2. Directly measure transcription factor activity in defined subpopulations 3. Model the transcriptional regulation of dopaminergic cell identity 4. Test model predictions by reprogramming astrocytes in vitro 5. Use the model predictions to reprogram astrocytes directly in vivo in striatum 6. Verify the outcome by single-cell RNA-seq of reprogrammed astrocytes, and comparison with the cell atlas Expected outcomes include: •"A detailed atlas of human ventral midbrain based on single-cell RNA-seq •"A mechanistic model of integration, by transcription factors, of patterning signals to specify dopaminergic fate •"Demonstration of direct in vivo reprogramming of astrocytes to dopaminergic neurons

Popular science description

I detta projekt vill vi uteckla en ny behandlingsmetod för parkinsons sjukdom, där patientegna astorcyter omprogrammeras på plats i striatum till dopamin-neuron. För att uppnå detta vill vi 1. Kartlägga ventrala mitthjärnan hos människa med single-cell RNA sekvensering 2. Direkt mäta transkriptionsfaktorers aktivitet i definierade cellpopulationer 3. Modellera och identifiera relevanta faktorer som kan omprogrammera celler till dopamin-neuron 4. Testa modellens förutsägelser in vitro genom att omprogrammera astrocyter 5. Testa modellen in vivo (i mus) genom att med virus programmera om astrocyter i striatum 6. Verifiera att omprogrammerade astrocyter in vivo förvandlats till dopamin-neuron, med single-cell RNA-seq