Bioengineering Self-Organizing Signaling Centers to Control Embryoid Body Pattern Elaboration

Area of Science:

• Developmental Biology
• Synthetic Biology
• Stem Cell Biology

Background:

• Multicellular systems exhibit self-organization, crucial for embryonic development, involving pattern formation and symmetry breaking.
• Artificial stem cell-derived structures like embryoid bodies (EBs) show self-organization but lack the control seen in vivo.
• Improved tools are needed to direct self-organization in engineered stem cell models.

Purpose of the Study:

• To develop a synthetic biology platform for user-defined control over self-organization in stem cell-derived structures.
• To engineer HEK-293 cells to induce specific developmental patterns within EBs.

Main Methods:

• Overexpressed P-cadherin (Cdh3) in HEK-293 cells to form clusters on EBs.
• Engineered these cells to produce mouse WNT3A for localized signaling.
• Coaggregated engineered HEK-293 cells with EBs to observe self-organization dynamics.

Main Results:

• Cdh3-expressing HEK-293 cells formed localized clusters on EBs.
• Localized WNT3A production triggered polarized Wnt/β-catenin pathway activation in EBs.
• Induced nascent mesoderm specification, pattern elaboration, and symmetry breaking within EBs.

Conclusions:

• Synthetic biology offers a platform to engineer self-organizing coaggregates.
• Localized WNT3A delivery can direct developmental patterning in stem cell structures.
• This approach advances the creation of more realistic synthetic developmental models.