Genetic-code-expanded cell-based therapy for treating diabetes in mice

Chen, Chao; Yu, Guiling; Huang, Yujia; Cheng, Wenhui; Li, Yuxuan; Sun, Yi; Liu, Tao

doi:10.1038/s41589-021-00899-z

Cited by 30 publications

(26 citation statements)

References 52 publications

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“…In contrast to the quick rise of insulin in response to a glucose surge in native β-cells, it may take more time to synthesize and secret insulin proteins after the increase of blood glucose. Although such designer cells are able to normalize glucose homeostasis in T1D mouse models, an ideal β-cell-mimetic designer cell is expected to control insulin protein translation (Chen et al, 2022) or secretion.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, a variety of synthetic biology-inspired approaches are developed to deliver therapeutic agents on demand by using triggercontrolled gene switches (Auslander and Fussenegger, 2013). Promising results have been generated in engineered non-islet cells that trigger the expression of gene of interest (GOI) (e.g., insulin) in response to glucose, electronic, light or non-canonical amino acids (Xie et al, 2016;Shao et al, 2017;Ye et al, 2017;Krawczyk et al, 2020;Wu et al, 2020;Li et al, 2021;Chen et al, 2022;Zhou et al, 2022). However, these single-trigger-controlled insulin expression systems remain risky in clinical application.…”

Section: Introductionmentioning

confidence: 99%

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A glucose-blue light AND gate-controlled chemi-optogenetic cell-implanted therapy for treating type-1 diabetes in mice

Wu²,

Zhao³

et al. 2023

Front. Bioeng. Biotechnol.

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Exogenous insulin therapy is the mainstay treatment for Type-1 diabetes (T1D) caused by insulin deficiency. A fine-tuned insulin supply system is important to maintain the glucose homeostasis. In this study, we present a designed cell system that produces insulin under an AND gate control, which is triggered only in the presence of both high glucose and blue light illumination. The glucose-sensitive GIP promoter induces the expression of GI-Gal4 protein, which forms a complex with LOV-VP16 in the presence of blue light. The GI-Gal4:LOV-VP16 complex then promotes the expression of UAS-promoter-driven insulin. We transfected these components into HEK293T cells, and demonstrated the insulin was secreted under the AND gate control. Furthermore, we showed the capacity of the engineered cells to improve the blood glucose homeostasis through implantation subcutaneously into Type-1 diabetes mice.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A glucose-blue light AND gate-controlled chemi-optogenetic cell-implanted therapy for treating type-1 diabetes in mice

Wu²,

Zhao³

et al. 2023

Front. Bioeng. Biotechnol.

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“…To ensure efficient delivery of POSH, we demonstrated compact co-delivery of the necessary components in a single plasmid as well as delivery as an all-in-one chimeric form. In contrast to a recently developed strategy that provides control in the translational step by incorporating non-canonical amino acids in the protein sequence ( 40 ), our post-translational control system works in the natural cell environment without the need for any exogenous supplement, other than the inducer. Although post-translational control strategies have recently been developed that allow timely control of protein secretion ( 20–22 , 41 ), they are limited to chemical inducers and have so far been employed only in vitro.…”

Section: Discussionmentioning

confidence: 99%

Design of programmable post-translational switch control platform for on-demand protein secretion in mammalian cells

Mansouri

Ray

Franko

et al. 2022

Nucleic Acids Research

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The development of novel strategies to program cellular behaviors is a central goal in synthetic biology, and post-translational control mediated by engineered protein circuits is a particularly attractive approach to achieve rapid protein secretion on demand. We have developed a programmable protease-mediated post-translational switch (POSH) control platform composed of a chimeric protein unit that consists of a protein of interest fused via a transmembrane domain to a cleavable ER-retention signal, together with two cytosolic inducer-sensitive split protease components. The protease components combine in the presence of the specific inducer to generate active protease, which cleaves the ER-retention signal, releasing the transmembrane-domain-linked protein for trafficking to the trans-Golgi region. A furin site placed downstream of the protein ensures cleavage and subsequent secretion of the desired protein. We show that stimuli ranging from plant-derived, clinically compatible chemicals to remotely controllable inducers such as light and electrostimulation can program protein secretion in various POSH-engineered designer mammalian cells. As proof-of-concept, an all-in-one POSH control plasmid encoding insulin and abscisic acid-activatable split protease units was hydrodynamically transfected into the liver of type-1 diabetic mice. Induction with abscisic acid attenuated glycemic excursions in glucose-tolerance tests. Increased blood levels of insulin were maintained for 12 days.

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“…More than 300 ncAAs have been genetically incorporated into proteins, providing powerful tools for investigating protein structures and functions. [1][2][3]7,[77][78][79][80][81][82][83][84][85] To date, utilizing these ncAAs in the context of Genetic Code Expansion has required both exogenous feeding and good membrane permeability of chemically-synthesized ncAAs. Cell membranes are poorly permeable to ncAAs with charged, highly hydrophobic, or hydrophilic structures.…”

Section: Discussionmentioning

confidence: 99%

Unleashing the Potential of Noncanonical Amino Acid Biosynthesis for Creation of Cells with Site-Specific Tyrosine Sulfation

Chen

Jin

Zhang

et al. 2022

Preprint

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Incorporation of noncanonical amino acids (ncAAs) into proteins holds great promise for modulating the structure and function of those proteins and for influencing evolutionary dynamics in organisms. Despite significant progress in improving the efficiency of translational machinery needed for incorporating ncAAs, exogenous feeding of high concentrations of chemically-synthesized ncAAs, especially in the case of polar ncAAs, is required to ensure adequate intracellular ncAA levels. Here, we report the creation of autonomous cells, both prokaryotic and eukaryotic, with the ability to biosynthesize and genetically encode sulfotyrosine (sTyr), an important protein post-translational modification with low membrane permeability. We discovered the first enzyme catalyzing tyrosine sulfation, sulfotransferase 1C1 from Nipponia nippon (NnSULT1C1), using a sequence similarity network (SSN). The unique specificity of NnSULT1C1 for tyrosine has been systematically explored using both bioinformatics and computational methods. This NnSULT1C1 was introduced into both bacterial and mammalian cells so as to yield organisms capable of biosynthesizing high levels of intracellular sTyr. These engineered cells produced site-specifically sulfated proteins at a higher yield than cells fed exogenously even with the highest level of sTyr reported in literature. We have used these autonomous cells to prepare highly potent thrombin inhibitors with site-specific sulfation. By enhancing ncAA incorporation efficiency, this added ability of cells to biosynthesize ncAAs and genetically incorporate them into proteins greatly extends the utility of genetic code expansion methods.

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Genetic-code-expanded cell-based therapy for treating diabetes in mice

Cited by 30 publications

References 52 publications

A glucose-blue light AND gate-controlled chemi-optogenetic cell-implanted therapy for treating type-1 diabetes in mice

A glucose-blue light AND gate-controlled chemi-optogenetic cell-implanted therapy for treating type-1 diabetes in mice

Design of programmable post-translational switch control platform for on-demand protein secretion in mammalian cells

Unleashing the Potential of Noncanonical Amino Acid Biosynthesis for Creation of Cells with Site-Specific Tyrosine Sulfation

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