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

The authors acknowledge Marion Johnson and all the members of the Morsut, McCain, Li, Khademhosseini Lab for insightful discussions and suggestions on the project. The authors acknowledge their family and friends that support them always and in particular for the times of this work that took place during the COVID-19 pandemic. Research reported in this publication was supported by NIGMS of the National Institutes of Health under award number R35GM138256 (L.M.); the National Science Foundation award number CBET-2145528 Faculty Early Career Development Program (L.M.); NSF RECODE from CBET-2034495 (M.L.M., L.M.); USC Department of Stem Cell Biology and Regenerative Medicine Startup Fund (L.M.); Wellcome Trust, Leap HOPE (L.M.), Viterbi Center for CIEBOrg (L.M., M.L.M.). T.H. acknowledges support from the Ruth L. Kirschstein National Research Service Award T32HL069766 and the UCLA Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, Research Award. Fellowships for students: CIRM fellowship for S.D., Fellowships from BME Department for the first year for M.G., N.C., and S.D. Grace True, Finacy Jin for technical support for the project. S.L. acknowledges the support of the Innovation Award from the UCLA Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, and grants (GM143485 and NS126918) from the National Institute of Health. GFP structures used in Figs. 1A, F and 2A, D created with BioRender.com. Marvin was used for drawing and displaying chemical structures, substructures, and reactions, Marvin 19.7.0, 2019, ChemAxon (http://www.chemaxon.com).

The source data file at the following link contains the quantified data points for each data graph in the figures. https://doi.org/10.6084/m9.figshare.25648026. The bulk and single-nuclei RNA sequencing data generated in this study have been deposited in the NIH GEO database under accession code GSE269404. Plasmids generated in this study are available on Addgene. Source data are provided with this paper.

The ImageJ macro used for the generation of scrambled pixel images for Fig. 3 is provided in the supplemental information file.

The technology transfer office of USC with the authors have filed patent disclosures with the technology described here; LM is an inventor on a previous synNotch patent for applications in cancer cell therapy licensed to Gilead; M.L.M. is an inventor on a patent on gelatin hydrogels licensed to Emulate. The remaining authors declare no competing interests.