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Local source of FGF8 specifies a fronto-temporal axis in engineered human brain organoids (PolarOids)

ABSTRACT: Local source of FGF8 specifies a fronto-temporal axis in engineered human brain organoids (PolarOids)

PROVIDER: PRJNA733039 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR14664005.fastq.gz	Fastqsanger.gz
	SRR14664006.fastq.gz	Fastqsanger.gz
	SRR14664007.fastq.gz	Fastqsanger.gz
	SRR14664008.fastq.gz	Fastqsanger.gz
	SRR14664009.fastq.gz	Fastqsanger.gz

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Local source of FGF8 specifies a fronto-temporal axis in engineered human brain organoids (PolarOids)

Project description:The establishment of the fronto-temporal axis in the developing human cortex is achieved by temporary signaling activity coming from secondary organizers. However, studying this process remains challenging due to the limited access to fetal material and inaccuracy of animal models in mimicking human-specific aspects. Although human brain organoids provide a potential solution to study this, their lack of axial organization does not supply the appropriate positional information needed to recapitulate detailed patterning processes in situ. Here, we develop a method that combines a cell line expressing the morphogen FGF8 fused to organoids engineered with PDMS moulds representing the exact length of the developing brain, resulting in the formation of polarized cortical organoids, or PolarOids. Thanks to a dose-sensitive reporter for anterior cortical differentiation, we carefully titrate and time the fusion dynamics to elicit a gradient in the patterned tissue while preserving neural activity as assessed through electrophysiology. We show that our localized FGF8 source is sufficient to trigger a distance-dependent transcriptional signature that matches fronto-temporal cortical fates in vivo. Lastly, we use these PolarOids to model a disease-relevant FGFR3 mutation and describe the transcriptional loss of axial gradient formation underlying a dramatic cortical phenotype observed in patients. As such, Polaroids provide a powerful tool to model hitherto neglected human diseases caused by defects in antero-posterior patterning in the brain

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Effect of FGF8 on bovine granulosa cells

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Specification of a rostro-caudal axis in cortical assembloids through a polarized source of FGF8 (scRNA-Seq)

Project description:Current organoid protocols are widely used to mechanistically study cortical development, evolution and brain pathologies. While these systems generate all major cortical cell types in histologically distinct compartments, they lack cortical area topography, which is initiated by gradients of signaling molecules released by signaling organizers that establish transcriptionally distinct domains along the rostro-caudal axis of the developing cortex. Here, we engineer human cortical assembloids that combine an organizer-like structure expressing FGF8 with an elongated organoid design to enable the controlled modulation of FGF8 signaling along the longitudinal organoid axis. We demonstrate that individual polarized cortical assembloids (polCA) mount a position-dependent transcriptional program that matches the in vivo rostro-caudal gene expression patterns. Importantly, positional identity is lost in polCA carrying a mutation in the FGFR3 gene associated with temporal lobe malformations and intellectual disability in humans. Similar to in vivo, this program is accompanied by transcriptional divergence of radial glia cells and excitatory neurons and their segregation along the longitudinal axis of individual organoids. Thus, this method reproduces in vivo cortical topography in individual organoids and enables the analysis of area-related phenotypes underlying human disorders.

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Specification of a rostro-caudal axis in cortical assembloids through a polarized source of FGF8 (bulk RNA-Seq)

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Transcription profiling of animal caps from Danio rerio embryos expressing GFP or vhnf1 and treated with FGF8 or BSA protein

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Fgf8 E18.5 hypomorph versus control mouse fetal lungs

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