1. Engineering programmable material-to-cell pathways via synthetic notch receptors to spatially control differentiation in multicellular constructs.
- Author
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Garibyan, Mher, Hoffman, Tyler, Makaske, Thijs, Do, Stephanie, Wu, Yifan, Williams, Brian, March, Alexander, Cho, Nathan, Pedroncelli, Nicolas, Lima, Ricardo, Soto, Jennifer, Jackson, Brooke, Santoso, Jeffrey, Khademhosseini, Ali, Thomson, Matt, Li, Song, McCain, Megan, and Morsut, Leonardo
- Subjects
Receptors ,Notch ,Cell Differentiation ,Tissue Engineering ,Animals ,Humans ,Signal Transduction ,Mice ,Extracellular Matrix ,Fibroblasts ,Extracellular Matrix Proteins ,Ligands ,Tissue Scaffolds ,Muscle ,Skeletal ,Endothelial Cells ,HEK293 Cells - Abstract
Synthetic Notch (synNotch) receptors are genetically encoded, modular synthetic receptors that enable mammalian cells to detect environmental signals and respond by activating user-prescribed transcriptional programs. Although some materials have been modified to present synNotch ligands with coarse spatial control, applications in tissue engineering generally require extracellular matrix (ECM)-derived scaffolds and/or finer spatial positioning of multiple ligands. Thus, we develop here a suite of materials that activate synNotch receptors for generalizable engineering of material-to-cell signaling. We genetically and chemically fuse functional synNotch ligands to ECM proteins and ECM-derived materials. We also generate tissues with microscale precision over four distinct reporter phenotypes by culturing cells with two orthogonal synNotch programs on surfaces microcontact-printed with two synNotch ligands. Finally, we showcase applications in tissue engineering by co-transdifferentiating fibroblasts into skeletal muscle or endothelial cell precursors in user-defined micropatterns. These technologies provide avenues for spatially controlling cellular phenotypes in mammalian tissues.
- Published
- 2024