Induced Neural Progenitor Specification from Human Pluripotent Stem Cells by a Refined Synthetic Notch Platform

Area of Science:

• Developmental Biology
• Stem Cell Biology
• Neuroscience

Background:

• Studying human brain and central nervous system (CNS) development in vitro is challenging.
• Human pluripotent stem cells (hPSCs) offer a model for early CNS development.
• Existing protocols often fail to replicate embryonic signaling centers, like the floor plate, crucial for neural tube patterning.

Purpose of the Study:

• To develop a cell-guided differentiation method using a synthetic Notch (synNotch) receptor platform.
• To precisely regulate Sonic Hedgehog (SHH) production for controlled cell fate specification.
• To improve upon existing synNotch configurations for robust signaling in hPSC cocultures.

Main Methods:

• Utilized a synthetic Notch (synNotch) receptor platform for cell-guided differentiation.
• Engineered refined designs of membrane-bound green fluorescent protein (GFP) ligands for enhanced receptor activation.
• Co-cultured ligand-presenting hPSCs ('senders') with synNotch-hPSCs ('receivers') to induce SHH production.

Main Results:

• Identified limitations in standard orthogonal synNotch configurations for hPSC signaling.
• Developed enhanced synNotch ligands enabling efficient receptor activation in hPSC receivers.
• Demonstrated that the revised synNotch system drives SHH production and generates floor plate-like cells.

Conclusions:

• The enhanced synNotch platform provides a robust method for cell-guided differentiation of hPSCs.
• This system successfully mimics floor plate signaling, a key event in neural tube development.
• The platform holds potential for synthetic morphogenesis studies to understand human CNS development paradigms.