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

Mher Garibyan; Tyler Hoffman; Thijs Makaske; Stephanie K Do; Yifan Wu; Brian A Williams; Alexander R March; Nathan Cho; Nicolas Pedroncelli; Ricardo Espinosa Lima; Jennifer Soto; Brooke Jackson; Jeffrey W Santoso; Ali Khademhosseini; Matt Thomson; Song Li; Megan L McCain; Leonardo Morsut

doi:10.1038/s41467-024-50126-1

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.

Authors

Mher Garibyan^#^{1

2

3}, Tyler Hoffman^#⁴, Thijs Makaske^{1

2

5}, Stephanie K Do³, Yifan Wu⁴, Brian A Williams⁶, Alexander R March^{1

2}, Nathan Cho³, Nicolas Pedroncelli⁴, Ricardo Espinosa Lima⁴, Jennifer Soto⁴, Brooke Jackson⁴, Jeffrey W Santoso³, Ali Khademhosseini^{4

7}, Matt Thomson⁶, 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. mlmccain@usc.edu.
¹¹ Alfred E. Mann Department of Biomedical Engineering, USC Viterbi School of Engineering, University of Southern California, Los Angeles, CA, USA. mlmccain@usc.edu.
¹² Department of Stem Cell Biology and Regenerative Medicine, Keck School of Medicine of USC, University of Southern California, Los Angeles, CA, USA. morsut@usc.edu.
¹³ Eli and Edythe Broad Center, University of Southern California, Los Angeles, CA, 90033, USA. morsut@usc.edu.
¹⁴ Alfred E. Mann Department of Biomedical Engineering, USC Viterbi School of Engineering, University of Southern California, Los Angeles, CA, USA. morsut@usc.edu.

^# Contributed equally.

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.

MeSH terms

Animals
Cell Differentiation*
Endothelial Cells / cytology
Endothelial Cells / metabolism
Extracellular Matrix / metabolism
Extracellular Matrix Proteins / genetics
Extracellular Matrix Proteins / metabolism
Fibroblasts / cytology
Fibroblasts / metabolism
HEK293 Cells
Humans
Ligands
Mice
Muscle, Skeletal / cytology
Muscle, Skeletal / metabolism
Receptors, Notch* / metabolism
Signal Transduction*
Tissue Engineering* / methods
Tissue Scaffolds / chemistry

Substances

Receptors, Notch
Extracellular Matrix Proteins
Ligands

Abstract

MeSH terms

Substances

Grants and funding