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Hierarchical mechanisms for transcription factor-mediated reprogramming of fibroblasts to neurons

ABSTRACT: Hierarchical mechanisms for transcription factor-mediated reprogramming of fibroblasts to neurons

PROVIDER: PRJNA188177 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR653912_1.fastq.gz	Fastqsanger.gz
	SRR653912_2.fastq.gz	Fastqsanger.gz
	SRR653913_1.fastq.gz	Fastqsanger.gz
	SRR653913_2.fastq.gz	Fastqsanger.gz
	SRR653914_1.fastq.gz	Fastqsanger.gz

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Hierarchical mechanisms for transcription factor-mediated reprogramming of fibroblasts to neurons

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Hierarchical mechanisms for transcription factor-mediated reprogramming of fibroblasts to neurons

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Project description:Direct lineage reprogramming is a promising approach for human disease modeling and regenerative medicine, with poorly understood mechanisms. Here, we reveal a hierarchical mechanism in the direct conversion of fibroblasts into induced neuronal (iN) cells mediated by the transcription factors Ascl1, Brn2, and Myt1l. Ascl1 acts as an "on-target" pioneer factor by immediately occupying most cognate genomic sites in fibroblasts. In contrast, Brn2 and Myt1l do not access fibroblast chromatin productively on their own; instead, Ascl1 recruits Brn2 to Ascl1 sites genome wide. A unique trivalent chromatin signature in the host cells predicts the permissiveness for Ascl1 pioneering activity among different cell types. Finally, we identified Zfp238 as a key Ascl1 target gene that can partially substitute for Ascl1 during iN cell reprogramming. Thus, a precise match between pioneer factors and the chromatin context at key target genes is determinative for transdifferentiation to neurons and likely other cell types.

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Project description:Recently, direct reprogramming between divergent lineages has been achieved by introducing cell-fate-determining transcription factors. This progress may provide alternative cell resources for drug discovery and regenerative medicine. However, the genetic manipulation may limit the future application of these approaches. In this study, we identified a novel small-molecule cocktail that directly converted fibroblasts into neuronal cell fate with a high yield up after 16-days of induction. After a further maturation stage, these chemically-induced neurons (CiNs) possessed neuron-specific expression patterns, generated action potentials and formed functional synapses. Gene expression profiling revealed the activation of neuronal specific genes in the early stage of small molecule treatment. Overall, our findings prove the principle of chemically-induced direct reprogramming of somatic cell fates across germ layers without genetic manipulation, and show that cell fate can be manipulated through disrupting initial cell program and activating target cell master genes with pure chemicals. Total of 15 samples were analyzed, including mouse fibroblasts, mouse cortical primary neurons and chemically-induced neurons by different duration of chemical induction (Day0, Day4, Day8, Day19) and different small-molecule cocktail (FICB, FICB-1)

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