Open AccessRewiring of endogenous signaling pathways to genomic targets for therapeutic cell reprogramming
Author(s) -
Krzysztof Krawczyk,
Leo Scheller,
Hyojin Kim,
Martin Fussenegger
Publication year - 2020
Publication title -
nature communications
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.559
H-Index - 365
ISSN - 2041-1723
DOI - 10.1038/s41467-020-14397-8
Subject(s) - jurkat cells , reprogramming , transactivation , microbiology and biotechnology , nfat , hek 293 cells , biology , signal transduction , mapk/erk pathway , smad , cell , gene expression , t cell , receptor , immune system , transcription factor , gene , genetics
Rewiring cellular sensors to trigger non-natural responses is fundamental for therapeutic cell engineering. Current designs rely on engineered receptors that are limited to single inputs, and often suffer from high leakiness and low fold induction. Here, we present Generalized Engineered Activation Regulators (GEARs) that overcome these limitations by being pathway-specific rather than input-specific. GEARs consist of the MS2 bacteriophage coat protein fused to regulatory or transactivation domains, and work by rerouting activation of the NFAT, NFκB, MAPK or SMAD pathways to dCas9-directed gene expression from genomic loci. This system enables membrane depolarization-induced activation of insulin expression in β-mimetic cells and IL-12 expression in activated Jurkat cells, as well as IL-12 production in response to the immunomodulatory cytokines TGFβ and TNFα in HEK293T cells. Engineered cells with the ability to reinterpret the extracellular milieu have potential for applications in immunotherapy and in the treatment of metabolic diseases.