Engineering programmable material-to-cell pathways via synthetic notch receptors to spatially control differentiation in multicellular constructs

  • Nat Commun. 2024 Jul 13;15(1):5891. doi: 10.1038/s41467-024-50126-1.
Mher Garibyan  #  1  2  3 ,  Tyler Hoffman  #  4 ,  Thijs Makaske  1  2  5 ,  Stephanie K Do  3 ,  Yifan Wu  4 ,  Brian A Williams  6 ,  Alexander R March  1  2 ,  Nathan Cho  3 ,  Nicolas Pedroncelli  4 ,  Ricardo Espinosa Lima  4 ,  Jennifer Soto  4 ,  Brooke Jackson  4 ,  Jeffrey W Santoso  3 ,  Ali Khademhosseini  4  7 ,  Matt Thomson  6 ,  Song Li  4  8  9 ,  Megan L McCain  10  11 ,  Leonardo Morsut  12  13  14
Affiliations
  • 1. Department of Stem Cell Biology and Regenerative Medicine, Keck School of Medicine of USC, University of Southern California, Los Angeles, CA, USA.
  • 2. Eli and Edythe Broad Center, University of Southern California, Los Angeles, CA, 90033, USA.
  • 3. Alfred E. Mann Department of Biomedical Engineering, USC Viterbi School of Engineering, University of Southern California, Los Angeles, CA, USA.
  • 4. Department of Bioengineering, University of California Los Angeles, Los Angeles, CA, USA.
  • 5. Utrecht University in the lab of Prof. Dr. Lukas Kapitein, Los Angeles, CA, 90024, USA.
  • 6. Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA.
  • 7. Terasaki Institute for Biomedical Innovation (TIBI), Los Angeles, CA, 90024, USA.
  • 8. Broad Stem Cell Center, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
  • 9. Jonsson Comprehensive Cancer Center, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
  • 10. Department of Stem Cell Biology and Regenerative Medicine, Keck School of Medicine of USC, University of Southern California, Los Angeles, CA, USA. [email protected].
  • 11. Alfred E. Mann Department of Biomedical Engineering, USC Viterbi School of Engineering, University of Southern California, Los Angeles, CA, USA. [email protected].
  • 12. Department of Stem Cell Biology and Regenerative Medicine, Keck School of Medicine of USC, University of Southern California, Los Angeles, CA, USA. [email protected].
  • 13. Eli and Edythe Broad Center, University of Southern California, Los Angeles, CA, 90033, USA. [email protected].
  • 14. Alfred E. Mann Department of Biomedical Engineering, USC Viterbi School of Engineering, University of Southern California, Los Angeles, CA, USA. [email protected].
  • # Contributed equally.
PMID: 39003263 DOI: 10.1038/s41467-024-50126-1
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.

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