Gaurav Singh Rathore

PhD student from the Kirkeby Lab

Titel: Mapping early human neural development in an in vitro model of human brain development using Single Cell RNAseq

Abstract

The complexity and size of the human brain is unique. However, due to limited availability of human fetal tissue, we have surprisingly little validated knowledge about the developmental processes which specify human brain regions and neuronal subtypes. Here, we tissue-engineered a model (MiSTR) of human neural tube regionalisation based on human embryonic stem cells (hESCs) and microfluidic cell culturing. By exposing differentiating hESCs to a WNT signalling gradient we mimicked early rostro-caudal neural patterning, and with >80% reproducibility generated a tissue with progressive caudalisation from forebrain over midbrain to hindbrain. Single-cell transcriptomics revealed that rostrocaudal organisation was established already at 24 hours of differentiation, before expression of neural markers. Moreover, the transcriptomic hallmarks of rostro-caudal MiSTR organisation accurately recapitulated gene expression patterns of the early rostro-caudal neural plate in mouse embryos. By implementing large scale single cell RNAseq on the cells from MiSTR tissue at different time points and regions , we aim to produce a comprehensive spatio-temporal map of human neural subtype specification, spanning an unprecedented variety of human neural lineages. For this we have sequenced approx 250K cells and build a web based MiSTR development atlas depcting gene expression and neural lineage trajectories.MiSTR thereby represents a novel in vitro model of human neurodevelopment to deconstruct and systematically analyse gene regulatory networks responsible for neural tube patterning.