Design principles for generating robust gene expression patterns in dynamic engineered tissues

Recapitulating native tissue organization is a central challenge in regenerative medicine as it is critical for generating functional tissues. One strategy to generate engineered tissues with predictable and appropriate organization is to mimic the gene expression patterning process that organizes tissues in the developing embryo. In a developing embryo, correct organization is accomplished by tissue patterning via the generation of temporal and spatial patterns of gene expression coupled with, and leading to, extensive cellular re-organization. Methods to pattern gene expression in vitro could therefore provide both better models for understanding the cellular and molecular events taking place during tissue morphogenesis and novel strategies for engineering tissues with more realistic and complex architectures. While a few attempts have been made to genetically pattern tissues in vitro, these do not produce sharp predictable patterning. In both the embryo and an in vitro tissue, patterning often occurs during extensive cell re-organization but how the dynamics of gene induction and cell re-distribution interact to impact the final outcome of patterning and ultimately tissue organization is not known. Understanding this relationship and the system parameters that dictate robust pattern formation is critical for engineering genetic patterning in vitro to organize artificial tissues. We set out to identify key requirements for pattern formation by patterning gene expression in vitro in sheets of re-distributing cells using a drug-inducible gene expression system and patterned drug delivery to mimic morphogen gene induction. Based on our experimental observations, we develop a mathematical model that allows us to identify and experimentally verify the conditions under which generation of sharp gene expression patterns is possible in vitro. Our results highlight the importance of coordinating gene induction dynamics and cellular movement in order to achieve robust pattern formation.