Dissecting Gene Regulatory Networks Governing Human Cortical Cell Fate

Area of Science:

• Neuroscience
• Developmental Biology
• Genetics

Background:

• Human cortical neurogenesis is a complex process involving radial glia (RG) neural stem cells.
• Transcription factor (TF) circuits are crucial for balancing RG self-renewal and differentiation into diverse neural cell types.
• Functional studies are needed to understand TF roles in human RG lineage progression.

Purpose of the Study:

• To investigate the roles of 44 transcription factors (TFs) in human cortical neurogenesis using a primary culture system and CRISPR interference (CRISPRi) screening.
• To identify novel TFs and effector genes involved in regulating neural stem cell fate and lineage progression.
• To explore conserved mechanisms of RG lineage plasticity across primates and uncover roles in neurodevelopmental disorders.

Main Methods:

• Established a human primary cell culture system for sensitive discrimination of cell fate dynamics.
• Applied single-cell clustered regularly interspaced short palindromic repeats interference (CRISPRi) screening to assess the impact of 44 TFs.
• Analyzed transcriptional and cell fate consequences of TF perturbations during cortical neurogenesis.

Main Results:

• Identified novel TFs, including ZNF219 (represses differentiation), NR2E1, and ARX, with critical roles in human cortical neurogenesis.
• Discovered convergent effector genes downstream of TFs linked to neurodevelopmental and neuropsychiatric disorders.
• Uncovered conserved mechanisms of RG lineage plasticity across primates and a postmitotic role for ARX in interneuron (IN) specification.

Conclusions:

• The study provides a functional framework for dissecting regulatory networks governing cell fate during human neurogenesis.
• Novel TFs and effector genes were identified, offering new insights into the genetic programs underlying cortical development.
• Findings highlight conserved mechanisms and potential links to human brain disorders, paving the way for future research.