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

• DOI: 10.1021/acssynbio.4c00742
• Publication Date: 2025-05-06T15:39:57Z
• Authors: Catherine A. Hamann; Andrew Kjar; Hyosung Kim; Alan J. Simmons; Hannah J. Brien; Cheryl I. Quartey; Bonnie L. Walton; Ken S. Lau; Ethan S. Lippmann; Jonathan M. Brunger
• Keywords: Synthetic Biology; Progenitor

Historically, studying the development of brain and central nervous system (CNS) tissues has been challenging. Human pluripotent stem cell (hPSC) technology allowed for in vitro reconstitution relevant, early trajectories by using small molecules recombinant proteins to guide differentiation cells toward relevant CNS phenotypes. However, many these protocols fail recapitulate cell-guided programs intrinsic embryonic development, particularly signaling centers that emerge within neural tube during formation. Located on ventral end tube, floor plate acts as one such center pattern dorsal/ventral axis secreting morphogen Sonic Hedgehog (SHH). Here, we present a method synthetic Notch (synNotch) receptor platform regulate SHH production subsequent fate specification. We show widely used configuration orthogonal synNotch ligand green fluorescent protein (GFP) mounted platelet-derived growth factor receptor-β transmembrane chassis does not allow robust artificial synNotch-hPSCs ("receivers") cocultured with ligand-presenting hPSCs ("senders"). discovered refined designs membrane-bound GFP-ligand efficient activation hPSC receivers. A variant this enhanced drives sender:hPSC receiver cocultures gives rise plate-like types seen development. This revised potential morphogenesis studies designed uncover key paradigms human