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BMP, MEK and WNT modulation with NGN2 expression for rapid neuron differentiation from hiPSC amenable to regional patterning [BMWi_freeze]

ABSTRACT: BMP, MEK and WNT modulation with NGN2 expression for rapid neuron differentiation from hiPSC amenable to regional patterning [BMWi_freeze]

PROVIDER: PRJNA1065918 | ENA |

REPOSITORIES: ENA

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			Action	DRS
		Other
	SRR27596534.fastq.gz	Fastqsanger.gz
	SRR27596535.fastq.gz	Fastqsanger.gz
	SRR27596537.fastq.gz	Fastqsanger.gz
	SRR27596539.fastq.gz	Fastqsanger.gz

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BMP, MEK and WNT modulation with NGN2 expression for rapid generation of hiPSC-derived neurons amenable to regional patterning [snRNA-seq]

Project description:hiPSC are a promising tool for studying neurological diseases and developing therapies for neurodegenerative diseases. Differentiation of hiPSC into neurons can be achieved by dual SMAD inhibition (dSMADi) or induced NGN2 overexpression (“iNGN2”). Starting directly from hiPSC, iNGN2 shortens the time to a neuronal stage but leads to neurons partially resembling peripheral or posterior fates while dSMADi more faithfully recapitulates forebrain development. To modify the iNGN2 paradigm, we applied an accelerated induction paradigm that is dependent on inhibition of BMP, MEK and WNT pathways (“BMWi”), to commit hiPSC into a rostral neuroectoderm fate before iNGN2. The resulting neurons showed strong expression of telencephalic and cortical markers, with decreased levels of peripheral and posterior marker genes compared to iNGN2 alone. The resulting cortical neurons are suitable for a tau aggregation assay. Furthermore, we could demonstrate that during BMWi treatment, the cells are amenable to additional regional patterning cues. This allowed the generation of neurons from different regions of the CNS and PNS, which will significantly facilitate in vitro modeling of a range of neurodevelopmental and neurodegenerative disorders.

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BMP, MEK and WNT modulation with NGN2 expression for rapid neuron differentiation from hiPSC amenable to regional patterning [BMWi_freeze]

Project description:hiPSCs aid in studying neurological diseases and developing therapies. Interventions, such as dual SMAD inhibition or NGN2 overexpression (“iNGN2”), can direct hiPSCs towards neurons. Starting directly from hiPSCs, iNGN2 shortens the time to a neuronal stage but leads to neurons resembling peripheral or posterior fates. We applied an accelerated induction paradigm that is only dependent on inhibition of BMP, MEK and WNT pathways (“BMWi”), to commit iPSCs before to rostral neuroectoderm. The resulting neurons showed strong expression of telencephalic, cortical markers, with decreased levels of peripheral and posterior marker genes compared to iNGN2 alone. The resulting cortical neurons are suitable for a tau aggregation assay. Furthermore, we could demonstrate that during BMWi treatment, the cells are amenable to regional patterning clues. This allowed the generation of neurons from different regions of the CNS and PNS, which will significantly improve existing in vitro models for neurodegenerative disorders.

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